References of "Sacheli, Rosalie"
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See detailMicroRNA Targeting of CoREST controls polarization of migrating cortical neurons
Volvert; Prévot, Pierre-Paul; Close, Pierre ULg et al

in Cell Reports (2014), 7(4), 1168-83

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See detailEVALUATION OF THE RAPID DETECTION OF ST-17 AND ST-1 GROUP B STREPTOCOCCI USING A MICROFLEX MALDI-TOF MS (BRUKER)
MEEX, Cécile ULg; SACHELI, Rosalie ULg; DESCY, Julie ULg et al

Poster (2014, May)

Objectives Clearly associated to neonatal meningitis, Group B streptococci (GBS) classified as sequence type-17 (ST-17) are defined as the “highly virulent” clone amongst GBS. The aim of this study was to ... [more ▼]

Objectives Clearly associated to neonatal meningitis, Group B streptococci (GBS) classified as sequence type-17 (ST-17) are defined as the “highly virulent” clone amongst GBS. The aim of this study was to evaluate an easy and rapid method, recently described to detect ST-17 and ST-1 GBS, based on distinguishing peak-shifts present on the protein spectrum of these 2 sequence types, using a Microflex (Bruker) matrix-assisted laser desorption/ionization time of flight mass spectrometer (MALDI-TOF MS). Methods This study was performed on 67 multi locus sequence typed (MLST) GBS originated from the Belgian and Czech National Reference Centers, including 18 ST-17 and 16 ST-1. After culture on blood agar, an ethanol/formic acid extraction was performed on each strain. Each extract was spotted once on a target plate, overlaid with 1 µl alpha-cyano-4-hydroxycinnamic acid matrix and further analysed by a Microflex MALDI-TOF MS. One spectrum per isolate was recorded, 240 laser shots being recorded for each spectrum. The spectra were further analysed using a Bruker prototype software, and 2 logarithmic values, one for ST-17 and one for ST-1, calculated from the intensities of the present and absent peaks, were obtained for each strain. If >0, this value indicated the presence of the specific sequence type. In a second step, the test was repeated on each strain with discordant result when compared with MLST. Results Compared with MLST method, the first analysis of the strains gave poor results, leading to very low sensitivities (77.8% for ST-17 and 50% for ST-1) but rather good specificities (85.7% for ST-17 and 98.0% for ST-1). After repeating the analysis on the strains with discordant result, sensitivity, 100% and 93.8%, and specificity, 87.8% and 98.0%, for ST-17 and ST-1 respectively were highly improved. Conclusion Since ST-17 and ST-1 GBS both show distinguishing peak-shifts on their protein spectrum, as described by Lartigue et al., the distinction of these 2 sequence types is now possible by MALDI-TOF MS. To our knowledge, this study is the first describing this application on a Microflex MS using a software to classify the strains. The observed results are promising but, given to the variability of the logarithmic value given by the software, the need to perform several measures on a same strain seems to be essential. After optimization of the analysis procedure, this rapid, easy and cheap method could be used to precociously detect ST-17 among GBS isolated from prenatal screenings, allowing a better follow up of the colonized mothers and a closer monitoring of their newborns. We would like to thank the Bruker Company which allowed us to evaluate the prototype software they have developed. [less ▲]

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See detailAssessment of pfcrt 72-76 haplotypes eight years after chloroquine withdrawal in Kinshasa, Democratic Republic of Congo
Mvumbi, Dieudonné; BOREUX, Raphaël ULg; SACHELI, Rosalie ULg et al

in Malaria Journal (2013), 12

BACKGROUND: In 2001, the World Health Organization (WHO) has recommended the use of artemisinin-based combination therapy (ACT) as the first-line treatment of uncomplicated malaria cases, as monotherapies ... [more ▼]

BACKGROUND: In 2001, the World Health Organization (WHO) has recommended the use of artemisinin-based combination therapy (ACT) as the first-line treatment of uncomplicated malaria cases, as monotherapies had become ineffective in many parts of the world. As a result, the Democratic Republic of Congo (DRC) withdrew chloroquine (CQ) from its malaria treatment policy in 2002 and an artesunate (AS)-amodiaquine (AQ) combination became the ACT of choice in DRC in 2005. AQ-resistance (AQR) has been reported in several parts of the world and mutations in codons 72-76 of the Plasmodium falciparum chloroquine-resistance transporter (pfcrt) gene have been strongly correlated with resistance, especially mutations encoding the SVMNT haplotype. This haplotype was first identified in Southeast Asia and South America but was recently reported in two African countries neighbouring DRC. These facts raised two questions: the first about the evolution of CQ resistance (CQR) in DRC and the second about the presence of the SVMNT haplotype, which would compromise the use of AQ as a partner drug for ACT. METHODS: A total of 213 thick blood films were randomly collected in 2010 from a paediatric clinic in Kinshasa, DRC. Microscopy controls and real-time polymerase chain reaction (RT-PCR) were performed for Plasmodium species identification. Haplotypes of the pfcrt gene were determined by sequencing. RESULTS: The K76T mutation was detected in 145 out of 198 P. falciparum-positive samples (73.2%).In these 145 resistant strains, only the CVIET haplotype was detected. CONCLUSIONS: This study is the first to assess the molecular markers of resistance to CQ and AQ after the introduction of ACT in DRC. The results suggest first that CQR is decreasing, as wild-type pfcrt haplotypes were found in only 26.8% of the samples and secondly that the SVMNT haplotype is not yet present in Kinshasa, suggesting that AQ remains valid as a partner drug for ACT in this region. [less ▲]

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See detailDNA fingerprinting using Diversilab system for genotyping characterization of Microsporum audouinii and Trichophyton violaceum
SACHELI, Rosalie ULg; DIMO, Lauryl; GRAIDE, Hélène ULg et al

in Mycoses (2013, October 01), 56(Supplement S3), 99

Objectives: To investigate the epidemiological determinants responsible for the high number of anthropophilic dermatophytes received by the National Reference Center for Mycosis of Liege (NRCL) during the ... [more ▼]

Objectives: To investigate the epidemiological determinants responsible for the high number of anthropophilic dermatophytes received by the National Reference Center for Mycosis of Liege (NRCL) during the year 2012. To perform a genotypic characterization by the Diversilab® system focusing on the two main isolated species, Microsporum audouinii and Trichophyton violaceum. To present a preliminary study preceding the national survey launched in 2013. Methods: A total of 51 strains of M. audouinii (50 clinical + 1 reference (ref.) strains) and 15 strains of T. violaceum (14 clinical + 1 ref. strain) originating from different locations through Belgium were included in the study. The fungal strains were first cultivated on Malt agar, then sub-cultured in Sabouraud liquid medium (Fluka). The grown mycelium was processed for DNA extraction following recommendations of the manufacturer (Ultra Clean® DNA Microbial isolation kit, MoBio laboratories). Genotypic analysis was performed using the DiversiLab® system (BioMérieux) for DNA fingerprinting and analysis. Results: Regarding M. audouinii, four different genotypic groups of strains were separated. Group 1 includes 11 strains and is only found in the Liège surroundings. Group 2 includes only one strain with little differences compared to group 1 and collected from the Liège area. These two groups may be related to each other. Group 3 contains 36 strains and the reference strain. This genotype is distributed in different Belgium locations. The last group, group 4, contains only 3 isolates sharing low similarities in comparison with the 3 other groups. Concerning T. violaceum, 6 different genotypic groups with a mixed geographical distribution were determined. Group 1 includes 8 clinical isolates and the ref. strain. The other five isolates are all different and seem not to be related to each other. Conclusion: The automated typing DiversiLab® system proved to be an easy and efficient method to investigate the molecular epidemiology of dermatophytes infections. Preliminary results of the study show that, through Belgium, several groups of isolates co-exist for M. audouinii and T. violaceum providing evidence of genetic heterogeneity. This variation can be related to acquired mutations due to environmental adaptation. Further investigations are necessary to better understand the impact of this genotypic variation. [less ▲]

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See detailSurveillance of serotypes and antimicrobial susceptibility profile in group B streptococcus (GBS) in Belgium
Melin, Pierrette ULg; SACHELI, Rosalie ULg; Sarlet, Gilles ULg et al

in Program and Abstract of the 53rd Intersciences Conference on Antimicrobial Agents and Chemotherapy. Washington, USA: ASM. (2013, September)

BACKGROUND Today GBS vaccines for prevention of severe neonatal disease through transplacental delivery of antibodies directly from immunized mothers are in advanced stage of development. For the ... [more ▼]

BACKGROUND Today GBS vaccines for prevention of severe neonatal disease through transplacental delivery of antibodies directly from immunized mothers are in advanced stage of development. For the introduction of any GBS vaccine there are urgent needs for pre and post vaccine enhanced surveillance studies of strains isolated from both neonatal diseases and vagino-rectal colonization of pregnant women. In Belgium, surveillance of invasive isolates is regularly done by the NRC. We report in this study a surveillance of colonizing isolates of GBS. METHODS In 2012, 344 GBS isolates were obtained from a Belgian surveillance for vagino-rectal colonization among pregnant women (max. 5 isolates/lab). Capsular types were determined by agglutination (Strep-B-latex, SSI, Denmark) and MICs by using a microdilution method (Sensititre) and Etest® (EUCAST interpretive criteria). Furthermore, for the erythromycin (E) resistant (R) isolates, the inducible (iMLS), constitutive (cMLS) and M phenotypes were assessed by a double-disk diffusion test. RESULTS Serotype III was the more common (27.6%) followed by V, II, Ia, Ib, IV, IX, VII and VI (18.1%, 16.4%, 13.4%, 7%, 4.7%, 2.5%, 0.8%, 0.5%) and 8.9% were non typable. All isolates were susceptible to penicillin ; 29% were R to E with a higher rate among serotypes IV and V (p<0.05). Among these E-R isolates, 93% exhibited the MLS phenotype (R to E and CC): 66% were cMLS with E MIC50>256 mg/L and 27% iMLS with E MIC50/MIC90 2/>8 mg/L. The M phenotype (R to E and S to C) was expressed by 7% of E-R isolates with E MIC50/MIC90 2/4 mg/L. CONCLUSION Compared with Belgian data relating to neonatal invasive strains (NRC reports) 1) Serotype V and II are more frequent and III less frequent among colonizing isolates 2) Prevalence of E-R is similar in percentage and phenotypes with the MLS R phenotype as major mechanism. Extended surveillance of both invasive and colonizing isolates is needed currently to prepare the follow-up in the future vaccine era. [less ▲]

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See detailImprovement of transport condition of swabs for group B streptococcal (GBS) screening
MELIN, Pierrette ULg; Dodémont, Magali; Sarlet, Gilles ULg et al

in Program and Abstract of the 53rd Intersciences Conference on Antimicrobial Agents and Chemotherapy. Washington, USA: ASM. (2013, September)

BACKGROUND For the screening-based strategy for prevention of perinatal GBS disease, CDC Guidelines as many others recommend use of appropriate transport media (Amies, Stuart, e.g.) and processing of ... [more ▼]

BACKGROUND For the screening-based strategy for prevention of perinatal GBS disease, CDC Guidelines as many others recommend use of appropriate transport media (Amies, Stuart, e.g.) and processing of specimen as soon as possible within 1 to 4 days. False negative cultures occur for several causes including lost of GBS viability during transport. Could Lim broth, recommended for the selective enrichment, and Granada tubes be used as transport media for swab? Simulating conditions of routine practice, Lim broth and Granada tubes, were evaluated in vitro as transport media. METHODS Tubes of 3 brands of Lim broth (Becton Dickinson, bioMérieux, Copan) and Granada tubes (bioMérieux) were inoculated with low inocula of 10-100 CFU of GBS. Each type of tubes was incubated at 4°C, room T° (RT) and 35°C. GBS were enumerated from each tube by subculture on blood agar after 1, 2, 3 and 4 days of storage at the different T°. All tests were processed in triplicates with 3 strains of GBS belonging to serotype Ia, III and V. RESULTS No difference of survival was observed between the 3 strains. T° had significant impact on GBS recovery for each type of tubes. At 4°C the viability was hardly sustained along the 4 days. At RT and 35°C, an increase >6 log of the inocula was observed. The increase of GBS density was sustained at least 4 days for the 3 brands of Lim broth. For the Granada broth, such increase was also observed but at day 3 for tubes incubated at 35°C, viability decreased and for some tubes, GBS subcultures were negative at day 3 or 4. CONCLUSION To improve sensitivity of GBS screening cultures, Lim broth could be recommended as a strong transport media and the advisable storage condition would be RT to 35°C up to 4 days. In this way, initiating selective enrichment culture at the time of collection of specimen would provide higher sensitivity even for low density of colonization. Transport at 4°C should be avoided in favour with RT to 35°C. Studies in clinical setting are expected. For Granada tubes, storage at RT was fine but improvement seemed restricted in time at 35°C as there was a loss of viability after 3 days. For Granada tubes, extended evaluation and delimitation of use are needed. [less ▲]

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See detailGene transfer in inner ear cells: a challenging race
Sacheli, Rosalie ULg; Delacroix, Laurence ULg; Van Den Ackerveken, Priscilla ULg et al

in Gene Therapy (2013), 20

Recent advances in human genomics led to the identification of numerous defective genes causing deafness, which represent novel putative therapeutic targets. Future gene-based treatment of deafness ... [more ▼]

Recent advances in human genomics led to the identification of numerous defective genes causing deafness, which represent novel putative therapeutic targets. Future gene-based treatment of deafness resulting from genetic or acquired sensorineural hearing loss may include strategies ranging from gene therapy to antisense delivery. For successful development of gene therapies, a minimal requirement involves the engineering of appropriate gene carrier systems. Transfer of exogenous genetic material into the mammalian inner ear using viral or non-viral vectors has been characterized over the last decade. The nature of inner ear cells targeted, as well as the transgene expression level and duration, are highly dependent on the vector type, the route of administration and the strength of the promoter driving expression. This review summarizes and discusses recent advances in inner ear gene-transfer technologies aimed at examining gene function or identifying new treatment for inner ear disorders. [less ▲]

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See detailExpression patterns of miR-96, miR-182 and miR-183 in the development inner ear
Sacheli, Rosalie ULg; Nguyen, Laurent ULg; Borgs, Laurence ULg et al

in Gene Expression Patterns (2009)

MicroRNAs (miRNAs) constitute a class of small non-coding endogenous RNAs that downregulate gene expression by binding to 3' untranslated region (UTR) of target messenger RNAs. Although they have been ... [more ▼]

MicroRNAs (miRNAs) constitute a class of small non-coding endogenous RNAs that downregulate gene expression by binding to 3' untranslated region (UTR) of target messenger RNAs. Although they have been found to regulate developmental and physiological processes in several organs and tissues, their role in the regulation of the inner ear transcriptome remains unknown. In this report, we have performed systematic in situ hybridization to analyze the temporal and spatial distribution of three miRNAs (miR-96, mR-182, and mR-183) that are likely to arise from a single precursor RNA during the development and the maturation of the cochlea. Strikingly we found that the expression of mR-96, mR-182 and mR-183 was highly dynamic during the development of the cochlea, from the patterning to the differentiation of the main cochlear structures. [less ▲]

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