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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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See detailMustard allergy: diagnostic and identification of specific allergens by immunoblotting
Courtois, Justine ULiege; BERTHOLET, Catherine ULiege; Cavalier, Etienne ULiege et al

Poster (2017, June 20)

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

Poster (2017, June 18)

Introduction Allergy to kiwi fruit is often associated with severe reactions in addition with oral allergy syndrome. Kiwi fruit matrix is very complex as it contains many allergenic proteins. We describe ... [more ▼]

Introduction Allergy to kiwi fruit is often associated with severe reactions in addition with oral allergy syndrome. Kiwi fruit matrix is very complex as it contains many allergenic proteins. We describe a clinical case of allergy to kiwi fruit (Actinidia deliciosa) in a woman presenting birch pollen allergy and recurrent urticaria. Objectives The diagnosis of kiwi fruit allergy is based on anamnesis, skin prick test (SPT) and specific IgE (sIgE) measurement to total kiwi fruit extract. Actually, the in vitro diagnostic tools cannot help the physician to define the precise kiwi allergen involved in the allergic reaction. Indeed, only one molecular allergen component is commercially available: Act d 8 (PR-10 protein, Birch Bet v 1-homologous). We aimed to adapt a 2D Western blot (WB) to get the molecular allergen sensitization profile of the patient. Afterwards, we used mass spectrometry (LC-MS/MS) to identify precisely the allergens. Methods We analyzed the serum of a 23 y.o. woman presenting a positive SPT to kiwi extract, low sIgE for kiwi extract (0.11kUA/L) and positive sIgE for Act d 8 (6.66 KUA/L). We extracted total Actinidia deliciosa proteins. Then, we separated proteins on the basis of their isoelectric point and molecular weight. The patient serum was analyzed by 2D WB in order to evaluate its sIgE reactivity against the different protein spots. Finally, the protein spots recognized by the patient sIgE were identified by LC-MS/MS. Results The patient sIgE sensitization profile showed 5 specific protein spots. Amongst them, we selected 2 spots and identified them by LC-MS/MS. The first spot situated around 25 kDa/pH5-6 was pointed out as Act d 1 (cystein protease). The second spot around 10 kDa/pH10 was identified as Act d 10 (LTP family). The result of sIgE against Act d 8 correlated perfectly with a third spot of 18 kDa/pH6-7. Conclusion We studied a birch pollen allergic woman presenting recurrent urticaria with low sIgE to kiwi extract but a positive SPT to kiwi. The 2D WB provided a sIgE profile showing multiple kiwi allergens. Amongst them, we confirmed Act d 8 which is associated with OAS in birch pollen allergic patients and identified Act d 1 and Act d 10, both frequently associated with severe reactions to food. We pointed out a potential role of Act d 1 and Act d 10 in the clinical symptoms of urticaria in this patient. Furthermore, we demonstrated superiority of 2D WB over the traditional diagnostic methods, unable to reach the same precision. [less ▲]

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See detailAllergie au froment : caractérisation de nouveaux allergènes par immunoblot 2D et spectrométrie de masse
Courtois, Justine ULiege; BERTHOLET, Catherine ULiege; LUKAS, Pierre ULiege et al

Poster (2017, April 26)

Introduction Le froment est une source allergénique complexe contenant un grand nombre de protéines qui sont difficiles à isoler et à identifier. Le but de ce projet est de développer une méthode de ... [more ▼]

Introduction Le froment est une source allergénique complexe contenant un grand nombre de protéines qui sont difficiles à isoler et à identifier. Le but de ce projet est de développer une méthode de diagnostic capable de relier les allergènes spécifiques à une clinique particulière via western blot (WB) 2D. Ensuite, la technique de spectrométrie de masse (LC-MS/MS) a été utilisée pour identifier ces allergènes. Méthodes Une extraction protéique suivie d’une séparation sur base du point isoélectrique et du poids moléculaire des protéines ont été réalisées. Vingt-cinq patients présentant des IgE spécifiques (IgEs) pour le froment ont été sélectionnés et classés en 3 groupes: anaphylaxie alimentaire induite par l’effort (AAIE), dermatite atopique (DA) et rhinite pollinique (RP). Leur sérum a été analysé par WB 2D afin d’évaluer la réactivité de leurs IgEs. Les profils de sensibilisation ont été comparés et les spots protéiques d’intérêt identifiés par LC-MS/MS. Résultats Des profils de sensibilisation spécifiques ont été identifiés pour chaque groupe. Pour le groupe AAIE, les spots protéiques sont situés à 35 kDa, pH6-9 et à 37-50 kDa, pH5-6. Pour le groupe DA, le profil est localisé autour de 50 kDa, pH9 ; 10 kDa, pH9 et à 20-75 kDa, pH3. Pour le groupe RP, le profil est situé vers 90 kDa, pH9. Les analyses en LC-MS/MS de ces spots protéiques ont mis en évidence différents allergènes potentiellement intéressants tels que tri a 26, tri a bA, tri a 34 et tri a tritin. Discussion L’étude présentée répond à une demande clinique afin de prévoir les réactions croisées, d’adapter les régimes et de limiter le risque de choc anaphylactique. Néanmoins, les résultats restent à corréler à l’anamnèse. Les manifestations cliniques dépendent des allergènes impliqués et de la voie d’exposition. Ce projet vise à mettre en évidence de nouveaux allergènes qui pourraient être associés à une symptomatologie clinique spécifique. Conclusion Des profils de sensibilisation spécifiques ont été identifiés pour les 3 groupes, mais l’identification de certains spots protéiques est encore en cours d’analyse. A terme, il sera possible d’établir un lien entre la symptomatologie spécifique et les allergènes responsables nouvellement identifiés. [less ▲]

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See detailDistinction between urine crystals by automated urine analyzer SediMAX conTRUST is specific but lacks of sensitivity.
Castiglione, Vincent ULiege; CAVALIER, Etienne ULiege; Diop, Coumba Souley ULiege et al

in Clinical Chemistry & Laboratory Medicine (2017)

Urine crystals are commonly encountered but few papers have been published about it while it is useful for patient’s follow-up, particularly the stone-formers. For the first time to our knowledge, we have ... [more ▼]

Urine crystals are commonly encountered but few papers have been published about it while it is useful for patient’s follow-up, particularly the stone-formers. For the first time to our knowledge, we have assessed the performance of an automated urine analyzer to identify and distinguish several types of crystals in a very large cohort of samples. We share in this report the results of method comparison of the SediMAX conTRUST with the polarized light microscopy which is the gold standard. We showed that the distinction between crystals and other urine elements was accurate. Negative predictive value was very good, but the positive predictive value was poor for most of crystals. The discrimination of different types of crystals between them by the automated urine analyzer still requires improvement to decrease reviewing rate and help clinician. [less ▲]

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See detailHunting for new wheat allergens : a 2D Immunoblot and mass spectrometry approach
Courtois, Justine ULiege; GADISSEUR, Romy ULiege; BERTHOLET, Catherine ULiege et al

Poster (2016, October 15)

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 detailCase report autour de cristalluries médicamenteuses atypiques
Castiglione, Vincent ULiege; GADISSEUR, Romy ULiege

Conference (2016, October 14)

We report here the cases of uncommon-shaped crystals in patient's urine. The first one is a 73years-old man with massive crystalluria observed in his microscopic urinalysis on Sedimax° (Menarini ... [more ▼]

We report here the cases of uncommon-shaped crystals in patient's urine. The first one is a 73years-old man with massive crystalluria observed in his microscopic urinalysis on Sedimax° (Menarini). Observed crystals had various shape (needle, stick, Feather...). The patient was hospitalized because of lactic acidosis due to suicide attempt by medication overdose. The following medication had been taken: prazepam, acetaminophen, tramadol and Triumeq°, a new anti-retroviral association of dolutegravir, abacavir and lamivudine. Infrared analysis of dryed urine didn"t allow to identify any common stone component, nor any medication. We further discuss about the possible implication of dolutegravir in crystalluria in this patient. Secondly, we show several N-acetylsulfamethoxazol (sulfamethoxazol metabolite) urine crystals detected in microscopic urinalysis of various patients. All had uncommon shape and were sometimes associated to acute kidney injury [less ▲]

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See detailALERBLOT: Interest of 2D Immunoblot for the diagnosis of allergy to wheat
BERTHOLET, Catherine ULiege; GADISSEUR, Romy ULiege; quinting, birgit et al

in Allergy (2016, June), 71

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See detailDiagnostic des allergies, les composants
GADISSEUR, Romy ULiege

Conference (2016, May 21)

Detailed reference viewed: 17 (4 ULiège)
See detailLa cristallurie
GADISSEUR, Romy ULiege

Conference (2016, May 12)

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See detailMise au point d’une nouvelle méthode de diagnostic des allergènes du froment par Western blot 2D
BERTHOLET, Catherine ULiege; GADISSEUR, Romy ULiege; Quinting, Birgit

in Revue Française d'Allergologie (2016, April), 56(3), 284

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See detailVitamin D status after a high dose of cholecalciferol in healthy and burn subjects
ROUSSEAU, Anne-Françoise ULiege; DAMAS, Pierre ULiege; LEDOUX, Didier ULiege et al

in Burns : Journal of the International Society for Burn Injuries (2015), 41(5), 1028-1034

Background: Burn patients are at risk of vitamin D (VD) deficiency and may benefit from its pleiotropic effects as soon as acute phase. Aim of this observational study was to assess effects of a ... [more ▼]

Background: Burn patients are at risk of vitamin D (VD) deficiency and may benefit from its pleiotropic effects as soon as acute phase. Aim of this observational study was to assess effects of a cholecalciferol (VD3) bolus on VD status in adult burn patients (Group B, GB) after admission, compared to healthy subjects (Group H, GH). Methods: Both groups received an oral dose of 100,000 IU VD3. Blood samples were collected before (D0) and 7 days (D7) after bolus to measure 250H-D, 1,25(OH)2-D, parathyroid hormone (PTH) and fibroblast growth factor 23 (FGF23). Albumin (ALB) and VD binding protein (DBP) were measured and used to calculate free 25OH-D level. Data were expressed as median (min–max) or proportions. Results: A total of 49 subjects were included: 29 in GH and 20 in GB. At D0, prevalence of VD deficiency was higher in GB: 25OH-D was 21.5 (10.1–46.3) ng/ml in GH vs 11 (1.8–31.4) ng/ml in GB. DBP and ALB were lower in GB. At D7, DBP was stable in both groups while ALB decreased in GB. 25OH-D increased by 66.6 (13.5–260.3)% in GH. In GB, changes in 25OH-D extended from 36.7% to 333.3% with a median increase of 33.1%. Similar changes were observed in each group for free 25OH-D. High FGF23 levels were observed in GB. Conclusions: This study highlighted the differences in VD status and in response to a high dose VD3 in burn patients when compared to healthy patients. Pitfalls in VD status assessment are numerous during acute burn care: 25OH-D measurement needs cautious interpretation and interest of free 25OH-D is still questionable. They should not prevent burn patients to receive VD supplements during acute care. Higher doses than general recommendations should probably be considered. [less ▲]

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See detailVitamine D et lithiases rénales : la pierre d'achoppement
CAVALIER, Etienne ULiege; GADISSEUR, Romy ULiege; Castiglione, Vincent ULiege

in Benhamou, Claude-Laurent; Souberbielle, Jean-Claude (Eds.) La vitamine D chez l'adulte (2015)

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See detailLes pierres aux reins
Castiglione, Vincent ULiege; GADISSEUR, Romy ULiege; CAVALIER, Etienne ULiege

Article for general public (2015)

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