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See detailSupplementation with 80,000 IU vitamin D3/month between November and April corrects vitamin D insufficiency without overdosing: Effect on serum 25-hydroxyvitamin D serum concentrations.
Tournier, Herve; Tran, Nicole; Dray, Nathalie et al

in Presse medicale (Paris, France : 1983) (in press)

INTRODUCTION: Vitamin D insufficiency, defined by a 25-hydroxyvitamin D (25OHD) serum concentration<20ng/mL, is highly frequent in the French general population, especially between November and April. The ... [more ▼]

INTRODUCTION: Vitamin D insufficiency, defined by a 25-hydroxyvitamin D (25OHD) serum concentration<20ng/mL, is highly frequent in the French general population, especially between November and April. The aim of this study was to evaluate whether 80,000 IU vitamin D3 every month during this period of the year was able to maintain a 25OHD level between 20 and 60ng/mL in apparently healthy subjects whatever their basal vitamin D status. METHODS: Ninety-eight subjects volunteered to receive an 80,000 IU vitamin D3 dose every month between November 2014 and April 2015. Serum 25OHD, calcemia and calciuria were measured just before the first dose (Month 0), just before the 4th dose (M4), and one month after the 6th dose (M7). RESULTS: At M0, 25OHD was 17.5+/-9.5ng/mL. Sixty subjects (61.2%) had a 25OHD<20ng/mL and 25 (25.5%) had a 25OHD<10ng/mL. 25OHD increased significantly at M4 (35.3+/-8.0ng/mL) and M7 (40.1+/-8.5) without change in calcemia and calciuria. At M4, 2 subjects had a 25OHD slightly below 20ng/mL (17.6 and 19.7ng/mL), and none had a concentration>60ng/mL. At M7, all had a serum 25OHD>20ng/mL and 2 subjects had a value slightly above 60ng/mL (62.1 and 63.2ng/mL). CONCLUSION: A monthly supplementation with 80,000 IU vitamin D3 between November and April corrected vitamin D insufficiency in subjects in whom it was initially very frequent, without overdosing. This protocol is simple, safe and costless, and can be easily implemented when physicians detect risk factors for hypovitaminosis D in patients for whom a 25OHD measurement is not indicated. [less ▲]

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See detailSpurious intraoperative PTH results observed with 2(nd), but not with 3(rd) generation PTH assays.
Cavalier, Etienne ULiege; SCHLECK, Marie-Louise ULiege; Souberbielle, Jean-Claude

in Clinica Chimica Acta (2018), 477

We report here an interference that has been observed with 2(nd) generation PTH assays (PTH2), but not with 3(rd) generation PTH ones (PTH3), during PTH monitoring occurring in a surgical intervention for ... [more ▼]

We report here an interference that has been observed with 2(nd) generation PTH assays (PTH2), but not with 3(rd) generation PTH ones (PTH3), during PTH monitoring occurring in a surgical intervention for the resection of a parathyroid adenoma. The patient was cured and calcium levels returned to normal the next day, but PTH did not decrease with PTH2 whereas it decreased by 50% with PTH3 assays during the intervention. The reason is probably a PTH fragment released by the parathyroid gland during surgery. This fragment possesses the C-terminal part of the peptide but lacks the first amino-acids and may thus be considered as a member of the non-(1-84) PTH fragments family. It has also a longer half-life than 1-84 PTH. To avoid reporting spurious results to the surgeons, we thus recommend using PTH3 assays for monitoring of intra-operative PTH. [less ▲]

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See detailSingle- versus multiple-sample method to measure glomerular filtration rate.
DELANAYE, Pierre ULiege; Flamant, Martin; Dubourg, Laurence et al

in Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association (2018)

Background: There are many different ways to measure glomerular filtration rate (GFR) using various exogenous filtration markers, each having their own strengths and limitations. However, not only the ... [more ▼]

Background: There are many different ways to measure glomerular filtration rate (GFR) using various exogenous filtration markers, each having their own strengths and limitations. However, not only the marker, but also the methodology may vary in many ways, including the use of urinary or plasma clearance, and, in the case of plasma clearance, the number of time points used to calculate the area under the concentration-time curve, ranging from only one (Jacobsson method) to eight (or more) blood samples. Methods: We collected the results obtained from 5106 plasma clearances (iohexol or 51Cr-ethylenediaminetetraacetic acid (EDTA)) using three to four time points, allowing GFR calculation using the slope-intercept method and the Brochner-Mortensen correction. For each time point, the Jacobsson formula was applied to obtain the single-sample GFR. We used Bland-Altman plots to determine the accuracy of the Jacobsson method at each time point. Results: The single-sample method showed within 10% concordances with the multiple-sample method of 66.4%, 83.6%, 91.4% and 96.0% at the time points 120, 180, 240 and >/=300 min, respectively. Concordance was poorer at lower GFR levels, and this trend is in parallel with increasing age. Results were similar in males and females. Some discordance was found in the obese subjects. Conclusion: Single-sample GFR is highly concordant with a multiple-sample strategy, except in the low GFR range (<30 mL/min). [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 detailSunscreens block cutaneous vitamin D production with only a minimal effect on circulating 25-hydroxyvitamin D
Libon, Florence ULiege; Courtois, Justine; LE GOFF, Caroline ULiege et al

in Archives of Osteoporosis (2017)

Summary A 50+ SPF sunscreen decreased significantly cutaneous vitamin D production following a single narrow-band (nb)UVB exposure, independently from the body surface area exposed. In contrast, the ... [more ▼]

Summary A 50+ SPF sunscreen decreased significantly cutaneous vitamin D production following a single narrow-band (nb)UVB exposure, independently from the body surface area exposed. In contrast, the circulating 25(OH)D3 levels were only minimally affected. It is probable that another endogenous source of precursors is selected when skin-originated precursors are lacking. Purpose Sunscreen use, highly advocated for preventing cutaneous carcinogenesis, is potentially leading to an aggravation of vitamin D deficiency with its consequences on bone health. The effect of sunscreens on circulating vitamin D levels remains debated. This study investigated the effect of sunscreen on cutaneous vitamin D production and circulating 25(OH)D3 levels, according to different body surface areas (BSA). Methods Vitamin D and 25(OH)D3 levels were measured in four groups exposed to a single nbUVB exposure on 9% (group I: head and hands), 23% (group II: head, hands and arms), 50% (group III: head, hands, arms and legs) and 96% (group IV: total body) of the body surface without and with a 50+ sun protection factor sunscreen. Results Sunscreen use decreased by 83, 88.3, 75.7 and 92.5% the cutaneous vitamin D production in groups I to IV, respectively, but only by 13.2, 10.5, 7.7 and 10.4% the values of circulating 25(OH)D3, correspondingly. Conclusions Although a 50+ sunscreen decreases significantly cutaneous vitamin D production following a single nbUVB exposure, and independently from the BSA, the circulating 25(OH)D3 levels were only minimally affected. This could be explained by a switch to another endogenous source of precursors. Short-term sunscreen use probably does not affect circulating vitamin D levels and hence does not increase the risk for osteoporosis. The effect of long-term sunscreen use remains however to be determined. [less ▲]

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See detailParaneoplastic secretion in parathyroid carcinoma: serum hCG as a tumor marker
VALDES SOCIN, Hernan Gonzalo ULiege; BETEA, Daniela ULiege; Daly, Adrian ULiege et al

in Acta Clinica Belgica (2017), 72(2), 5

Introduction Parathyroid carcinoma (PCa) is a rare presentation of primary hyperparathyroidism (PHPT), accounting for less than 1 % of cases. Differentiating parathyroid cancer from adenoma is clinically ... [more ▼]

Introduction Parathyroid carcinoma (PCa) is a rare presentation of primary hyperparathyroidism (PHPT), accounting for less than 1 % of cases. Differentiating parathyroid cancer from adenoma is clinically challenging. Rubin et al (2008) et al. suggested that urinary hCG might be a marker of disease progression in PCa. In this study, we aimed to investigate whether the hCG+β kit from Roche Diagnostics could distinguish PCa patients from PHPT. Material and methods We studied a series of 8 patients suffering from advanced PCa, referred to the CHU de Liege. A control group of 20 PHPT patients was used as comparative. hCG+β kit on Cobas (Roche Diagnostics) uses 2 monoclonal antibodies that recognize holo-hCG, nicked hCG, β-core fragment and free β-subunit. Limits of hCG detection and quantification are <0.1 and <0.6 mUI/mL. In non pregnant and postmenopausal women and in men, hCG (p95) is <1 (5.3), <7 mUI/mL (8.3) and <2 (2.6) mUI/mL, respectively. Results. The 8 PCa patients (3 women) presented high serum hCG values at: 1.29, 3.46, 5.7, 24.2, 31.2, 34.1, 36.5 and 164 mUI/mL. Values of 1.29 and 3.46 were obtained in 2 postmenopausal women. The lowest value was presented by the only still alive patient. There was a significant correlation (r=0.786; ρ <0.05) between hCG and PTH and a borderline correlation (r=0.750; ρ =0.05) between hCG and calcium concentrations. All PHP patients presented undetectable hCG values. Conclusions These results suggest that serum hCG might have the potential to discriminate between parathyroid adenomas and carcinomas, with a sensibility of 75% (6/8) and a specificity of 100% (0/20). The only patient still alive presented the lowest hCG values. If hCG could be predictive of PCa survival needs to be studied in a larger series of patients. [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 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 detailOral calcium load test for recurrent calcium stone-formers
Castiglione, Vincent ULiege; CAVALIER, Etienne ULiege

Poster (2017, November 24)

Background Calcium is the most frequent component of urinary stones, and hypercalciuria is the main risk factor in recurrent stone-formers. The oral calcium load test is a dynamical biological test that ... [more ▼]

Background Calcium is the most frequent component of urinary stones, and hypercalciuria is the main risk factor in recurrent stone-formers. The oral calcium load test is a dynamical biological test that determines the origin of hypercalciuria in order to optimize the treatment. However, there is little literature about it, and it seems to have lost popularity in daily practice, this why we studied a population of stone-formers who underwent the oral calcium load test. Methods Between 2013 and 2016, we prospectively recruited 117 recurrent calcium stone-formers. After 2 days of calcium restricted-diet, patients had urinary and blood sampling at baseline and 120 min after the intake of 1 g of calcium per os. Blood and urinary parameters were assessed during the dynamical test, including stone risk factors, calcium metabolism and bone evaluation. According to these results, patients were classified in three groups: resorptive, renal or absorptive hypercalciuria. Results First, 19 patients were diagnosed with normocalcemic primary hyperparathyroidism, assessed by inappropriate parathormone decrease (41.41±12.82 vs. 54.06±13.84% p<0.01) in regard to calcemia. The measurement of ionized calcium was mandatory in order to detect induced hypercalcemia after calcium intake. These patients also had higher beta-crosslaps, lower phosphate reabsorption threshold and lower distal third radius bone mineral density. The treatment of this first group of patient is the hyperparathyroïdectomy. Fasting hypercalciuria was present in 39 patients with urinary calcium >0.37mmol/mmol of creatinine, and without hyperparathyroidism, classified thus as renal hypercalciuria. The treatment of these patients should include adapted calcium intake and thiazids. The third group included 34 patients with absorptive hypercalciuria defined by the presence of delta urinary calcium/creatinine <0.6mmol/mmol between 0 and 120 min, and without any other significant abnormality. Finally, the test result was not reliable for 33 cases because of the absence of sufficient calcemia increase or when the cause of lithogenesis could not be clearly identified. Conclusions The oral calcium load test was successful for the identification of main metabolic conditions leading to urolithiasis, including normocalcemic primary hyperparathyroidism, and is useful to improve and personalize the treatment of stone-formers. [less ▲]

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See detailOral calcium load test for recurrent calcium stone-formers
Castiglione, Vincent ULiege; CAVALIER, Etienne ULiege

Conference (2017, November 24)

Background Calcium is the most frequent component of urinary stones, and hypercalciuria is the main risk factor in recurrent stone-formers. The oral calcium load test is a dynamical biological test that ... [more ▼]

Background Calcium is the most frequent component of urinary stones, and hypercalciuria is the main risk factor in recurrent stone-formers. The oral calcium load test is a dynamical biological test that determines the origin of hypercalciuria in order to optimize the treatment. However, there is little literature about it, and it seems to have lost popularity in daily practice, this why we studied a population of stone-formers who underwent the oral calcium load test. Methods Between 2013 and 2016, we prospectively recruited 117 recurrent calcium stone-formers. After 2 days of calcium restricted-diet, patients had urinary and blood sampling at baseline and 120 min after the intake of 1 g of calcium per os. Blood and urinary parameters were assessed during the dynamical test, including stone risk factors, calcium metabolism and bone evaluation. According to these results, patients were classified in three groups: resorptive, renal or absorptive hypercalciuria. Results First, 19 patients were diagnosed with normocalcemic primary hyperparathyroidism, assessed by inappropriate parathormone decrease (41.41±12.82 vs. 54.06±13.84% p<0.01) in regard to calcemia. The measurement of ionized calcium was mandatory in order to detect induced hypercalcemia after calcium intake. These patients also had higher beta-crosslaps, lower phosphate reabsorption threshold and lower distal third radius bone mineral density. The treatment of this first group of patient is the hyperparathyroïdectomy. Fasting hypercalciuria was present in 39 patients with urinary calcium >0.37mmol/mmol of creatinine, and without hyperparathyroidism, classified thus as renal hypercalciuria. The treatment of these patients should include adapted calcium intake and thiazids. The third group included 34 patients with absorptive hypercalciuria defined by the presence of delta urinary calcium/creatinine <0.6mmol/mmol between 0 and 120 min, and without any other significant abnormality. Finally, the test result was not reliable for 33 cases because of the absence of sufficient calcemia increase or when the cause of lithogenesis could not be clearly identified. Conclusions The oral calcium load test was successful for the identification of main metabolic conditions leading to urolithiasis, including normocalcemic primary hyperparathyroidism, and is useful to improve and personalize the treatment of stone-formers. [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 detailComparison of cardiac biomarkers fluctuation in runners of marathons, semi-marathons and untrained runners
Le Goff, Caroline ULiege; VRANKEN, Laura ULiege; Kaux, Jean-François ULiege et al

in European Journal of Sports Medicine (2017, September), 5(Supplement 1), 16

BACKGROUND: Regular exercise like running is one important part of the prevention program of cardiovascular disease. There are several studies on biomarker changes during marathons especially cardiac ... [more ▼]

BACKGROUND: Regular exercise like running is one important part of the prevention program of cardiovascular disease. There are several studies on biomarker changes during marathons especially cardiac biomarkers have been studied and mild to moderate elevations have been described as a results of a running exercise Exact underlying mechanism for these biomarker elevations reflecting physiological or even pathobiological changes is unknown and less trained athletes might exhibit a higher risk compared to well trained The aim of our study was to compare three cardiac biomarkers for ischemic condition (Troponin), cardiac stretch (natriuretic peptides) and fibrotic processes (Galectin3) were tested in different type of runners, trained marathon and semimarathon runners and untrained runners before, directly after and 3 hours after the running exercise. METHODS: 23 (mean age 41± 8.8 yo) marathon runners, 15 semimarathon runners(44.1±8.4yo) and 17 healthy sedentary subjects (37± 4.4 yo) (race of 10 km, <2h of sport/week). Blood samples were taken just before (T0), just after (T1) and 3 hours (T3) after the race, centrifuged, aliquoted and stored frozen at 80C before further analysis. The study was approved by the Ethical Committee of our University Hospital. The analyses were performed on the Abbott ARCHITECT i2000SR (Abbott Laboratories, Germany) for the hs cTnI, BNP and Gal3 and on the C8000 (Roche Diagnostics, Switzerland) for hscTnT and NTproBNP according to the manufacturer’s instructions for use. RESULTS: In all 3 running groups there is an increase of cardiac biomarkers Troponin I, BNP, Galectin3 and NTProBNP after completion of the physical exercise. Biomarkers increase is depending on the intensity and duration of the exercise and is higher in long distance marathon and semimarathon runners compared to the control group with a 1 hour run. Cardiac biomarker levels between trained marathon and semimarathon runners were not statistically different in the preexercise baseline samples for BNP, NTProBNP and Galectin3. Compared to untrained runners only Troponin I levels were higher in baseline sample of marathon runners (hscTnI, p<0.03) when compared to controls, cardiac Troponin T (hscTnT, p<0.29) was less significant. CONCLUSIONS: Our study demonstrates that exercises of different intensity can be associated with biochemical abnormalities that may reflect adverse consequences on cardiac structure as fibrosis and biology. With exeption of Troponin I there was no difference in the preexercise baselines samples. [less ▲]

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See detailA fast and simple method for simultaneous measurements of 25(OH)D, 24,25(OH)2D and the Vitamin D Metabolite Ratio (VMR) in serum samples by LC-MS/MS (powerpoint)
Fabregat Cabello, Neus ULiege; Farre Segura, Jordi ULiege; Huyghebaert, Loreen ULiege et al

Scientific conference (2017, September)

We present here a rapid, easy, reliable and cost-effective method for the quantification of 25-hydryxoyvitamin D2 and D3, epi-25-hydroxyyvitamin D3 and 24,25-dihydroxyvitamin D3 by LC-MS/MS in serum ... [more ▼]

We present here a rapid, easy, reliable and cost-effective method for the quantification of 25-hydryxoyvitamin D2 and D3, epi-25-hydroxyyvitamin D3 and 24,25-dihydroxyvitamin D3 by LC-MS/MS in serum samples. The proposed methodology has been strongly validated with both NIST and Labquality materials, obtaining mean intra-assay and inter-assay imprecision lower than 6 and 6.4% and mean recoveries within 95-104%. Besides we have compared satisfactorily samples from Vitamin D Standardization Program (n=80) with reference values and patient samples (n=281) with our reference LC-MS/MS method. The proposed methodology is prepared to be used in routine testing and permits the calculation of the Vitamin D Metabolite Ratio (VMR). [less ▲]

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See detailA fast and simple method for simultaneous measurements of 25(OH)D, 24,25(OH)2D and the Vitamin D Metabolite Ratio (VMR) in serum samples by LC-MS/MS
Fabregat Cabello, Neus ULiege; Farre Segura, Jordi ULiege; Huyghebaert, Loreen ULiege et al

in Clinica Chimica Acta (2017)

BACKGROUND: Rapid, easy and reliable measurement of the major vitamin D metabolites is required in order to fulfill the needs of a clinical routine laboratory. To overcome these challenges, we have ... [more ▼]

BACKGROUND: Rapid, easy and reliable measurement of the major vitamin D metabolites is required in order to fulfill the needs of a clinical routine laboratory. To overcome these challenges, we have developed and validated a LC-MS/MS method for the quantification of 25-hydroxyvitamin D2 and D3, epi-25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3. METHODS: Sample preparation was based on precipitation and centrifugation of 100 µL of patient serum, followed by injection into the LC-MS/MS system. Samples from Vitamin D Standardization Program (n=80) and patient samples (n=281) have been compared with a reference LC-MS/MS method. For the analytical validation NIST and Labquality quality control materials were used. RESULTS: Mean intra-assay and inter-assay imprecision were less than 6.0 and 6.4% and mean recoveries were within 95-104%. LOQ’s were 0.5 µg/L for 24,25(OH)2D3, 1.1 µg/L for 25(OH)D3 and epi-25(OH)D3 and 1.7 µg/L for 25(OH)D2. A 3% bias obtained between the proposed and the reference method satisfies Vitamin D Standardization Program recommendations. CONCLUSIONS: We present a rapid, easy, reliable and cost-effective method completely adequate for routine testing, which permits the measurement of the ratio of 24,25-dihydroxyvitamin D to 25-hydroxyvitamin D, Vitamin D Metabolite Ratio (VMR), in serum samples. [less ▲]

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