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See detailRaman chemical imaging, a new tool in kidney stone structure analysis: Case-study and comparison to Fourier Transform Infrared spectroscopy
Castiglione, Vincent ULiege; Sacre, Pierre-Yves ULiege; Cavalier, Etienne ULiege et al

Poster (2018, November 16)

The kidney stone’s structure might provide clinical information in addition to the stone composition. The Raman chemical imaging is a technology used for the production of two- dimension maps of the ... [more ▼]

The kidney stone’s structure might provide clinical information in addition to the stone composition. The Raman chemical imaging is a technology used for the production of two- dimension maps of the constituents’ distribution in samples. We aimed at determining the use of Raman chemical imaging in urinary stone analysis. Fourteen calculi were analyzed by Raman chemical imaging using a confocal Raman micro- spectrophotometer. They were selected according to their heterogeneous composition and morphology. Raman chemical imaging was performed on the whole section of stones. Once acquired, the data were baseline corrected and analyzed by MCR-ALS. Results were then compared to the spectra obtained by Fourier Transform Infrared spectroscopy. [less ▲]

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See detailDiagnostic accuracy of three automated urine analyzers compared to urine culture
Castiglione, Vincent ULiege; GADISSEUR, Romy ULiege; Bernard, Maxime ULiege et al

Poster (2018, November 16)

Urine culture is an important diagnosis method of urinary tract infection. However, it is time-consuming and results aren’t available quickly. Automated urine analyzers have been developed to screen urine ... [more ▼]

Urine culture is an important diagnosis method of urinary tract infection. However, it is time-consuming and results aren’t available quickly. Automated urine analyzers have been developed to screen urine samples more rapidly. The goal of this study was to compare three automated analyzers to urine culture: the Atellica UAS 800 (Siemens, Munich, Germany), the UF-4000 (Sysmex, Kobe, Japan) and the SediMAX (Menarini, Florence, Italy). We first validated each analyzer. We analyzed then 318 samples with the three analyzers within 2 hours after sample reception. An urine aliquot was collected before sediment analysis for bacteria culture. Ten microliters of un-centrifuged urine was inoculated on blood agar and CLED agar plates, then they were incubated aerobically at 36°C for 24 h. Bacteria count of each analyzer was compared to urine culture to determine diagnostic accuracy. The result was considered positive when the bacteria growth reached 104 CFU/ml. We also used the results of leukocytes and nitrites results from the strip measurement to improve the accuracy. The abilities of the UF-4000 to distinguish Gram positive (GP) from Gram negative (GN) bacteria, and of the UAS 800 to identify rod and cocci, were determined. [less ▲]

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See detailUrine sediment analysis: comparison of three automated analyzers to manual microscopy
Castiglione, Vincent ULiege; GADISSEUR, Romy ULiege; Bernard, Maxime ULiege et al

Poster (2018, November 16)

Urine microscopic analysis is an old method that reveals information about kidney health. Several automated analyzers, that are less time-consuming, are currently available, but manual microscopy is still ... [more ▼]

Urine microscopic analysis is an old method that reveals information about kidney health. Several automated analyzers, that are less time-consuming, are currently available, but manual microscopy is still the gold-standard method. The goal of this study was to validate and compare three automated analyzers to manual microscopy: the Atellica UAS800 (Siemens, Munich, Germany), the UF-4000 (Sysmex, Kobe, Japan) and the SediMax (Menarini, Florence, Italy). We first validated each analyzer. A total of 359 samples were analyzed with the three analyzers and with a manual microscope within 2 hours. Two trained reviewers used a microscope with bright field, contrast phase and polarized light to identify urine elements. The diagnostic accuracy was determined thanks to microscopy results. [less ▲]

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See detailComparison of three automated strip analyzers to Cobas 800 for the analysis of glucose, proteins, albumin and creatinine
Castiglione, Vincent ULiege; GADISSEUR, Romy ULiege; Bernard, Maxime ULiege et al

Poster (2018, November 16)

Urine glucose, proteins, albumin and creatinine are measured for the screening of diabetes and renal diseases. The automated strip analyzers are used for quick screening of large populations. The goal of ... [more ▼]

Urine glucose, proteins, albumin and creatinine are measured for the screening of diabetes and renal diseases. The automated strip analyzers are used for quick screening of large populations. The goal of this study was to compare a more accurate method, the Cobas 8000 (Roche, Bale, Switzerland) to three automated analyzers: the Clinitek Novus (Siemens, Munich, Germany), the UC-3500 (Sysmex, Kobe, Japan) and the AutionMax (Menarini, Florence, Italy). A total of 284 urine samples were prospectively collected for the comparison. The samples were analyzed on the three analyzers within 2 hours. Before the analysis of samples by each method, an aliquot was frozen at -80°C. All samples were then defrost and analyzed in one batch with the Cobas 8000 (Roche, Bale, Switzerland) within the month. The diagnostic accuracy was determined thanks to the results of the Cobas 8000. However, the creatinine and albumin cannot be assessed with the AutionMax. [less ▲]

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See detailClinical data on rare Sulfamethoxazole crystalluria assessed by Fourier Transform Infrared Spectrophotometry
CASTIGLIONE, Vincent ULiege; CAVALIER, Etienne ULiege; GADISSEUR, Romy ULiege

in Data in Brief (2018)

The data contained in this article are related to the article entitled “Case report: Uncommon Sulfamethoxazole Crystalluria” [1]. Sulfamethoxazole crystalluria is very rare and crystals identification is ... [more ▼]

The data contained in this article are related to the article entitled “Case report: Uncommon Sulfamethoxazole Crystalluria” [1]. Sulfamethoxazole crystalluria is very rare and crystals identification is complex [2], [3]. We identified seven patients with uncommon urine crystals that were composed of N-Acetyl-Sulfamethoxazole. Three of the patients developed an acute renal failure simultaneously to crystalluria. Hence, this article describes the method of crystals identification thanks to infrared spectroscopy. The relevant clinical data of patients, including medical history, drug dosage and urine parameters related to the crystalluria are presented. [less ▲]

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See detailIntérêt du dosage de la tryptase dans un service d'urgences
GADISSEUR, Romy ULiege

Scientific conference (2018, October 17)

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See detailUrine sediment analysis: comparison of three automated analyzers
Castiglione, Vincent ULiege; GADISSEUR, Romy ULiege

Conference (2018, October 09)

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See detailComment j'explore...Une protéinurie
Résimont, Guillaume ULiege; GADISSEUR, Romy ULiege; LUTTERI, Laurence ULiege et al

in Revue Médicale de Liège (2018), 73(10), 519-525

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See detailValidation of 24 blood gas analyzers, GEM 5000 Premier
GADISSEUR, Romy ULiege; MUSSO, Giuseppe ULiege; Cavalier, Etienne ULiege et al

Poster (2018, September 28)

Introduction: Point-of-care blood gas test results may help to take therapeutic decision by their immediate impact on patient care. Recently, a novel cartridge-type blood gas analyzer, the GEM Premier ... [more ▼]

Introduction: Point-of-care blood gas test results may help to take therapeutic decision by their immediate impact on patient care. Recently, a novel cartridge-type blood gas analyzer, the GEM Premier 5000 (IL-Werfen) was commercialized and 24 analyzers were installed at the University Hospital of Liège. One of them was installed in the main central laboratory, the 23 other ones were installed into 19 patient care units. Before the implementation, we evaluated the analytical performance of all the 24 GEM Premier 5000, for the determination of whole blood pH, pCO2 and pO2, electrolytes (Na+, K+, Cl−), ionized calcium (iCa2+), glucose, lactate and co-oximetry parameters ((total hemoglobin (tHb), oxyhemoglobin (O2Hb), carboxyhemoglobin (COHb), methemoglobin (metHb), deoxyhemoglobin (HHb)). Method: First, we evaluated the performance of the GEM Premier 5000 dedicated to the central lab, so-called “referent analyzer”, with 3 levels of External Quality Controls material (EQC RNA Medicals). CLSI EP5 recommends 2 replicates per run, 1 or 2 runs per day, for a minimum of 20 days. Nevertheless, we analyzed the 3 levels of EQC, 2 replicates per run, 2 runs per day during 5 consecutive days. Afterwards, on the whole 24 GEM Premier 5000, we analyzed aqueous QC material (Werfen GEM System Evaluator, level 1-2), 3 replicates within a single run, once per day, during 5 consecutive days. We determined the manufacturer's claim for Within-Run and Total precisions for each. Co-oxymetry parameters were not evaluated on 4 analyzers. Then, we compared the all 23 analyzers to the “referent analyzer” of the central lab. Therefore, for each parameter, we showed in a Youden diagram all the results obtained by 23 analyzers. The position of the acceptance ranges were shown graphically using the specifications for Acceptable(%) Root Mean Standard Deviation (RMSD) proposed by the German Guidelines for Quality (RILIBAK) for whole blood parameter, for different ranges of parameters. Results: The results showed a good correlation between analyzers excepted for some parameters. Lactate and MetHb: level 2 were often over-estimated when compared to “Reference Analyzer”. It could be explained by the fact that this IQC level contains very low Lactate and MetHb rates. The pO2 level2: some results were over estimated (random errors >< cassette reagents >< low values). The pO2 level1: over-estimated with 8 analyzers letting us think that the cassette reagent of our “Reference Analyzer” had a bias in the lower range. Conclusion: Performance evaluation of a large cluster of Blood Gas Analyzers is always a challenge for a Hospital. Accreditation is one of the main goal in each Belgian laboratory. Hospital Accreditation is also discussed in Belgium. This study shows an interesting approach to validate Blood Gas Analyzers for highlighting data. Based on our study results, we estimated that the evaluated instrument are a suitable blood gas analyzer for both POCT and laboratory use. [less ▲]

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See detailRaman Chemical Imaging, a new tool in kidney stone structure analysis: Case-study and comparison to Fourier Transform Infrared spectroscopy
Castiglione, Vincent ULiege; Sacre, Pierre-Yves ULiege; CAVALIER, Etienne ULiege et al

in PLoS ONE (2018), 13(8), 0201460

Background and objectives: The kidney stone’s structure might provide clinical information in addition to the stone composition. The Raman chemical imaging is a technology used for the production of two ... [more ▼]

Background and objectives: The kidney stone’s structure might provide clinical information in addition to the stone composition. The Raman chemical imaging is a technology used for the production of two-dimension maps of the constituents' distribution in samples. We aimed at determining the use of Raman chemical imaging in urinary stone analysis. Material and methods: Fourteen calculi were analyzed by Raman chemical imaging using a confocal Raman microspectrophotometer. They were selected according to their heterogeneous composition and morphology. Raman chemical imaging was performed on the whole section of stones. Once acquired, the data were baseline corrected and analyzed by MCR-ALS. Results were then compared to the spectra obtained by Fourier Transform Infrared spectroscopy. Results: Raman chemical imaging succeeded in identifying almost all the chemical components of each sample, including monohydrate and dihydrate calcium oxalate, anhydrous and dihydrate uric acid, apatite, struvite, brushite, and rare chemicals like whitlockite, ammonium urate and drugs. However, proteins couldn't be detected because of the huge autofluorescence background and the small concentration of these poor Raman scatterers. Carbapatite and calcium oxalate were correctly detected even when they represented less than 5 percent of the whole stones. Moreover, Raman chemical imaging provided the distribution of components within the stones: nuclei were accurately identified, as well as thin layers of other components. Conversion of dihydrate to monohydrate calcium oxalate was correctly observed in the centre of one sample. The calcium oxalate monohydrate had different Raman spectra according to its localization. Conclusion: Raman chemical imaging showed a good accuracy in comparison with infrared spectroscopy in identifying components of kidney stones. This analysis was also useful in determining the organization of components within stones, which help locating constituents in low quantity, such as nuclei. However, this analysis is time-consuming, making it more suitable for research studies rather than routine analysis. [less ▲]

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See detailElucidation de la structure de la lithiase urinaire par imagerie IR-TF
Coic, Laureen ULiege; Castiglione, Vincent ULiege; Sacre, Pierre-Yves ULiege et al

Poster (2018, May)

Les lithiases urinaires sont des concrétions minérales qui se forment au niveau des reins. En fonction de leur taille et de leur composition, leur présence peut être un signe avant-coureur de maladies ... [more ▼]

Les lithiases urinaires sont des concrétions minérales qui se forment au niveau des reins. En fonction de leur taille et de leur composition, leur présence peut être un signe avant-coureur de maladies graves. Elles sont généralement analysées par spectroscopie Infrarouge à transformée de Fourier (IR-TF). La structure de la lithiase apporte des informations de l’évolution de la lithiase au court du temps. Celle-ci a des conséquences cliniques qui permettent d’améliorer la qualité des soins apportés au patient. Or, cette structure n’est pas élucidable par l’analyse conventionnelle en IR-TF car la lithiase est broyée pour pouvoir l’analyser ne donnant donc qu’une information moyenne de la composition. Le couplage de l’imagerie avec la spectroscopie IR-TF apporte un réel intérêt pour élucider la structure de la lithiase urinaire car c’est une technique non destructive et relativement rapide (temps d’analyse moyen par lithiase < 3h). Pour réaliser l’étude, onze lithiases avec des compositions chimiques différentes ont été sélectionnées puis analysées par imagerie IR-TF en réflexion (DRIFTS). Les images ont été acquises avec une résolution latérale de 5.5 µm et une résolution spectrale de 4 cm-1. Les données hyperspectrales obtenues ont été prétraitées avec une correction de ligne de base de type Whittaker. Les spectres des différents composés et les cartes de distribution afférentes ont été obtenus par décomposition des images à l’aide de la MCR-ALS. Les images IR permettent d’identifier la totalité des composés présents dans les lithiases avec une corrélation spectrale de plus de 0,90 pour l’ensemble des molécules. Dans certains cas, l’imagerie permet de mettre en exergue la présence de composés qui n’avaient pas été détectés par spectroscopie IR conventionnelle. Les images obtenues par MCR-ALS permettent de discriminer l’oxalate de calcium monohydraté (Whewhellite) de l’oxalate de calcium dihydraté (Weddellite) qui étaient pour alors difficilement discernables par spectroscopie. L’identification des protéines est également possible lorsqu’elles sont présentes à plus de 10% de la surface de la lithiase. L’imagerie IR montre ainsi un fort potentiel dans l’évaluation qualitative de la composition et de la structure de la lithiase urinaire et pourrait être transposée à d’autres applications métabolomiques. [less ▲]

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See detailCase report: Uncommon Sulfamethoxazole Crystalluria
Castiglione, Vincent ULiege; CAVALIER, Etienne ULiege; GADISSEUR, Romy ULiege

in Clinical Biochemistry (2018)

We report seven Caucasian patients with unusual Sulfamethoxazole crystalluria on random urine sediment analysis. Crystals had very uncommon and various shapes and were identified thanks to infrared ... [more ▼]

We report seven Caucasian patients with unusual Sulfamethoxazole crystalluria on random urine sediment analysis. Crystals had very uncommon and various shapes and were identified thanks to infrared spectroscopy. An eGFR (MDRD) decrease was associated to crystalluria onset in three patients. [less ▲]

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See detailIntérêt du Western blot 2D dans les réactions croisées avec les insectes
Courtois, Justine ULiege; Tollenaere, Stéphanie; Quinting, Birgit et al

Poster (2017, December 01)

Introduction L’entomophagie est une alternative alimentaire qui devient, depuis quelques années, une pratique de plus en plus courante dans nos pays. Le but de notre étude est donc d’identifier les ... [more ▼]

Introduction L’entomophagie est une alternative alimentaire qui devient, depuis quelques années, une pratique de plus en plus courante dans nos pays. Le but de notre étude est donc d’identifier les potentielles réactions croisées entre les allergènes des crustacés, des acariens et des grillons (Gryllodes sigillatus). Matériel & Méthodes Nous avons sélectionné 12 patients présentant une allergie aux acariens et/ou aux crustacés sur base de leur positivité en IgE spécifiques (IgEs) dirigés contre deux tropomyosines, Der p 10 (acariens) et contre Pen a 1 (crevettes) dosées par ImmunoCAP250 (ThermoFisher Scientific). Ensuite, nous avons réalisé une extraction protéique totale à partir de grillons séchés dans le but de séparer les protéines selon leurs points isoélectriques et leurs poids moléculaires. Finalement, nous avons réalisé un Western blot (WB) 1D suivi de WB 2D afin de déterminer les allergènes moléculaires responsables d’un profil de sensibilisation pour chaque sérum de patients testés contre cet extrait protéique. Résultats & Discussion Le WB 1D a confirmé la réactivité des IgEs contre une protéine située aux environs de 37 kDa qui pourrait être soit la tropomyosine soit l’arginine kinase (AK) du grillon. Le WB 2D a également confirmé une sensibilisation contre une protéine située aux alentours de 37 kDa, pH 3-4, probablement la tropomyosine et/ou contre une protéine localisée aux environs de 37 kDa, pH 6-7, probablement l’AK. De plus, une zone a également été mise en évidence aux environs de 17,5 kDa, pH 4 qui pourrait être la troponine C, une autre protéine allergénique décrite. Conclusion Nos résultats préliminaires ont montré qu’il y avait, bel et bien, une réaction croisée entre les crustacés/acariens et les grillons. Les protéines en cause sont la tromomyosine et/ou l’AK ainsi que la troponine C. Ces hypothèses seront confirmées par spectrométrie de masse (LC-MS/MS). [less ▲]

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See detailShould we all eat insects ?
Courtois, Justine ULiege; Goddé, Marie; Tollenaere, Stéphanie et al

Poster (2017, November 25)

Introduction Entomophagy is an alternative food that has become more common in our countries during recent years. The aim of the study was to identify the potential cross-reactivity between the allergens ... [more ▼]

Introduction Entomophagy is an alternative food that has become more common in our countries during recent years. The aim of the study was to identify the potential cross-reactivity between the allergens of shrimps, House Dust Mites (HDM) and crickets. Material & Method We selected 3 patients (aged 7–18–27 y.o.) on the basis of their positive sIgE results against Der p 10 (from 16.9 to >100 KUA/L) and against Pen a 1 (from 14.3 to >100 KUA/L), two tropomyosins. Each patient had a diagnosis of both HDM allergy and food allergy to shrimps. We performed a total Gryllodes sigillatus protein extraction in order to separate the proteins on the basis of their isoelectric point and their molecular weight. Afterwards, we performed 1D and 2D Western blot (WB) to determine the molecular allergen sensitization profile of each patient serum to the extract. Results & Discussion The 1D WB confirmed the sIgE reactivity to a protein around 37 kDa that could be the Gryllodes’ tropomyosin. The 2D WB confirmed for the 3 patients’ sera a tropomyosin sensitization (around 37 kDa, pH 5-7). Furthermore, it showed for 1 out of 3 patients a sensitization to a protein around 17 kDa, pH 9 that could be troponin, another described allergenic protein. Conclusion Our preliminary results showed IgE cross-reactivities with the Gryllodes’ tropomyosin in 3 shrimp and HDM allergic patients with positive sIgE to Pen a 1 and Der p 10. One patient presented a sensitization to the Gryllodes’ troponin, but the identification of this protein should be confirmed by LC-MS/MS. [less ▲]

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See detailInterest of 2D Immunoblot and mass spectrometry in the diagnosis of wheat allergy
Courtois, Justine ULiege; BERTHOLET, Catherine ULiege; Cavalier, Etienne ULiege et al

Poster (2017, November 24)

Aims Wheat is a complex allergenic food containing a lot of proteins that are difficult to isolate and to identify. Hence we aimed to develop a diagnostic method linking specific allergenic 2D western ... [more ▼]

Aims Wheat is a complex allergenic food containing a lot of proteins that are difficult to isolate and to identify. Hence we aimed to develop a diagnostic method linking specific allergenic 2D western blot profiles to a particular clinical symptom in wheat allergy. Afterwards, we used mass spectrometry (LC-MS/MS) to identify molecular allergens. Methods A total protein extract of wheat seeds was separated on the basis of the isoelectric point and the molecular weight of the proteins. Twenty-five patients presenting positive specific IgE (sIgE) for wheat were classified into 3 different phenotypes: wheat dependent exercise induced anaphylaxis (WDEIA), atopic dermatitis (AD) and pollen rhinitis (PR). Their sera were analyzed by 2D immunoblotting on a standardized wheat seeds extract in order to evaluate their sIgE reactivity against the protein spots. Their sIgE sensitization profiles were compared and protein spots of interest were identified by LC-MS/MS. Results Specific sensitization profiles were identified for each phenotype group. For WDEIA, protein spots around 37 kDa (pH 6-9) and 37-50 kDa (pH 5-6) were identified. For AD, spots were observed around 50 kDa (pH 9), 10 kDa (pH 9) and 20 to 75 kDa (pH3). For PR, specific spots were situated around 90 kDa (pH 9). The LC-MS/MS analysis of these identified spots pointed out several potential interesting allergens: tri a 26, tri a bA, tri a 34, tri a tritin. Discussion Our study answers to the request of many allergists wishing to get an accurate diagnosis of wheat allergy in order to determinate the risk of cross-reaction, to adapt the diet and to limit the risk of anaphylactic choc. Nevertheless, it is necessary to consider the 2D immunoblot results with the medical history of each patient. Moreover, there are different clinical manifestations of wheat allergy depending on the involved allergen and the way of exposure: WDEIA, AD, PR or baker's asthma. The present project pointed out new wheat allergens that could be associated to a specific phenotype. The identification of further protein spots is still under investigation. Conclusion At this stage, specific sensitization profiles were identified for the 3 phenotype groups (WDEIA, DA, PR). The protein spots of interest detected by sIgE concern one or more allergens. Some wheat allergens were identified by LC-MS/MS. At the end of the study, it will be possible to establish a link between a specific symptomatology and the responsible allergens newly identified. [less ▲]

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See detailHow immunoblotting and mass spectrometry can help to diagnose mustard allergy
Courtois, Justine ULiege; BERTHOLET, Catherine ULiege; Cavalier, Etienne ULiege et al

Poster (2017, November 24)

Introduction We describe a clinical case study of severe allergy to a variety of mustard, Sinapis alba, in an adult patient without any previous food nor respiratory allergy history. Objectives The ... [more ▼]

Introduction We describe a clinical case study of severe allergy to a variety of mustard, Sinapis alba, in an adult patient without any previous food nor respiratory allergy history. Objectives The diagnosis of allergy to mustard is based on anamnesis, skin prick test and specific IgE (sIgE) measurement to total mustard extract. Actually, the in vitro diagnostic tools cannot help the physician to define the precise mustard allergens involved in the allergic reaction and are unable to support evaluation of potential cross-reactions. Indeed, no molecular allergen component is commercially available for mustard. We aimed to adapt a 2D immunoblot method to mustard. Afterwards, mass spectrometry (LC-MS/MS) was used to identify precisely the allergens bound to sIgE. Methods We analyzed the serum of a 37 y.o. man presenting a grade 2 reaction (facial quincke edema with respiratory distress) after eating food containing the mustard species Sinapis alba. He had positive sIgE results for mustard extract (0.62 KUA/L) and a positive realistic SPT to foods containing mustard. We extracted total proteins of Sinapis alba seed. The different proteins were separated based on their isoelectric point and their molecular weight. The patient serum was analyzed by 2D Western blot in order to evaluate its sIgE reactivity against the different protein spots. Finally, the protein spots recognized by the patient sIgE were precisely identified by LC-MS/MS. Results The patient sIgE sensitization profile showed three specific protein spots. The first protein spot was observed at 18 kDa and pH 5 to 6. A second protein spot was localized around 14 kDa and pH 5. Finally, the third protein spot was situated around 15 kDa and pH 7. The LC-MS/MS analysis of these 3 spots pointed out 2 allergens already described in mustard allergy: sin a 1 (2S-albumin) and sin a 2 (11S-globulin). Conclusion In this study, a 2D immunoblot provided a specific sensitization profile for a patient presenting a grade 2 allergy to mustard with low sIgE to total mustard extract and without any other history of allergy. The protein spots recognized by the sIgE concerned two main allergens identified by LC-MS/MS as sin a 1 and sin a 2. Those allergens are classified in the storage protein family which is associated to severe reactions to food and could be highly cross-reactive. We pointed out specific mustard allergens that could be associated to severe reactions such as facial quincke edema with respiratory distress. [less ▲]

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See detailRare Sulfamethoxazole Crystalluria – case report
Castiglione, Vincent ULiege; Diop, Coumba Souley ULiege; CAVALIER, Etienne ULiege et al

Poster (2017, November 24)

Case discussion We report here 7 Caucasian patients with very uncommon crystalluria. There were 3 female and 4 male of 54 to 86 years-old. Patients had no relevant medical record in common. However, they ... [more ▼]

Case discussion We report here 7 Caucasian patients with very uncommon crystalluria. There were 3 female and 4 male of 54 to 86 years-old. Patients had no relevant medical record in common. However, they all were hospitalized for different types of infections: three patients had urinary infection, two had osteitis, and the two last had sepsis. The patients had all been first treated with different antibiotherapy, which had then been replaced by cotrimoxazole after antibiogram. The administrated doses varied from 800mg to 4800mg a day of Sulfamethoxazole. Crystalluria description In all patients, the urine microscopic analysis revealed unusual crystals of various shapes including rectangles, thick parallelepipeds, truncated lozenges, spheroids, mushrooms, or “flowers”. Some crystals were incorrectly identified by the urinary sediment analyzer as uric acid, but we sought to determine them accurately. Most of the crystals were strongly birefringent and measured between 20 and 100µm. Urine pH varied from 5.0 to 6.5 on strip analysis. After urine centrifugation, we performed infrared spectrophotometry analysis on dried residue. In all cases, the infrared spectra allowed us to identify the N-acetyl-Sulfamethoxazole, the main metabolite of Sulfamethoxazole. Crystalluria was observed between 1 and 26 days after Sulfamethoxazole treatment initiation. The serum creatinine increased from 16% to 88% in 3 patients between the first day of Sulfamethoxazole treatment and the day of crystalluria. These considerations raised concern for drug implication in renal failure in some of these patients. Teaching points for the clinical condition Drug-induced kidney failure is well-known, but the direct precipitation of drug crystals into tubules is rare, and also probably under-evaluated. Even if Sulfamethoxazole tubular precipitation was probably not the main cause of renal failure in these cases, we suspect it could have played a role. N-Acteyl-Sulfamethoxazole can precipitate in urine in many uncommon crystals shapes that raise suspicion for drug nephrotoxicity. Automated urine analyzers may misidentify these rare crystals. Crystal’s recognition may be difficult even with polarized light microscopy. This is why they must be identified by infrared spectrophotometry to avoid misdiagnosis. These renal failures linked to Sulfamethoxazole precipitation are more susceptible to appear with high dosage of drug, hypovolemia and pre-existing renal failure. Hypoalbuminemia has also been described as a risk factor and was present in our four patients (between 26 to 39g/l, range laboratory: 43-54). Thus, prevention of Sulfamethoxazole precipitation consists in hydratation to maintain urine flow, and require adaptation of cotrimoxazole dosage in regard of renal function. Urine alkalinization (pH >7.0) is also possible in order to increase Sulfamethoxazole metabolite solubility. [less ▲]

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See detailRaman Chemical Imaging in Kidney Stone Analysis
Castiglione, Vincent ULiege; Sacre, Pierre-Yves ULiege; CAVALIER, Etienne ULiege et al

Poster (2017, November 02)

Background: The structure of kidney stones might provide clinical useful information in addition to the stone composition. The Raman chemical imaging (RCI) is a new technology used for the production of ... [more ▼]

Background: The structure of kidney stones might provide clinical useful information in addition to the stone composition. The Raman chemical imaging (RCI) is a new technology used for the production of two-dimensions maps of the constituents' distribution in samples. We aimed at determining the use of RCI in urinary stone analysis. Methods: Twelve calculi were analyzed by RCI using a confocal Raman microspectrophotometer. They were selected according to their heterogeneous composition and morphology. Prior to the analysis, samples were sliced and milled in order to detect the nucleus of the stones and having a smooth surface. RCI was performed on the whole section of stones. Once acquired, the data were baseline corrected and analyzed by MCR-ALS. Results were then compared to the spectra obtained by Fourier Transform Infrared spectroscopy, the gold standard method for the determination of urolithiasis composition. Results: RCI succeeded in identifying all the chemical components contained in each sample, including monohydrate and dihydrate calcium oxalate, anhydrous and dihydrate uric acid, apatite, struvite, brushite, whitlockite and ammonium urate. However, proteins couldn't be detected because of the huge autofluorescence background and the small concentration of these poor Raman scatterers. Carbapatite and calcium oxalate were correctly detected even when they represented less than 5 percent of the whole stones, allowing the detection of very small structures like Randall's plaques. Moreover, RCI provided the distribution of components within the stones. The nuclei were accurately identified, as well as thin layers of other components. Conversion of dihydrate to monohydrate calcium oxalate was correctly observed in the center of one sample. Conclusion: RCI showed a good accuracy in comparison with infrared spectroscopy in identifying components of kidney stones. In addition, RCI is nondestructive enabling the storage of samples. This analysis was also useful in determining the organization of components within stones, which help locating constituents in low quantity, such as nuclei. However, this analysis is time-consuming, which makes it more suitable for research studies rather than routine analysis. [less ▲]

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See detailEntomophagy : what about allergies ?
Courtois, Justine ULiege; Goddé, Marie; Cavalier, Etienne ULiege et al

Conference (2017, October 10)

Introduction The entomophagy is an alternative food that has become more common in our countries during recent years. The aim of this study was to identify the potential cross-reactivity between the ... [more ▼]

Introduction The entomophagy is an alternative food that has become more common in our countries during recent years. The aim of this study was to identify the potential cross-reactivity between the allergens of shrimps, House Dust Mites (HDM) and crickets (Gryllodes sigillatus). Material & Method We selected 3 patients (aged 7–18–27 y.o.) on the basis of their positive specific IgE (sIgE) results against Der p 10 (from 16.9 to >100 KUA/L) and against Pen a 1 (from 14.3 to > 100 KUA/L), two tropomyosins. Each patient had a diagnosis of both HDM allergy and food allergy to shrimps. We performed a total Gryllodes sigillatus protein extraction in order to separate the proteins on the basis of their isoelectric point and on their molecular weight. Afterwards, we performed 1D and 2D Western blot (WB) to determine the molecular allergen sensitization profile of each patient serum to this extract. Results & Discussion The 1D WB confirmed the sIgE reactivity to a protein around 37 kDa that could be the Gryllodes’ tropomyosin or the Gryllodes’ arginine kinase (AK). The 2D WB confirmed for the 3 patients’ sera a tropomyosin sensitization (around 37 kDa, pH 3-4) or an AK sensitization (around 37 kDa, pH 6-7). Furthermore, it showed for 1 out of 3 patients a sensitization to a protein around 17,5 kDa, pH 4 that could be troponin C, another described allergenic protein. Conclusion Our preliminary results showed IgE cross-reactivities with the Gryllodes’ tropomyosin or AK in 3 shrimp and HDM allergic patients with positive sIgE to Pen a 1 and Der p 10. One patient presented a sensitization to the Gryllodes’ troponin C, but the identification of this protein should be confirmed by mass spectrometry (LC-MS/MS). [less ▲]

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