Comparaison des tests de dépistage rapide et de la mesure de la protéinurie chez le chien

Les tests de dépistage des protéinuries utilisables par le vétérinaire praticien ont été établis pour une utilisation en médecine humaine. La détermination de leur valeur diagnostique chez le chien est nécessaire car ces tests ne sont pas validés dans cette espèce. Le but de cette étude a été de comparer une bandelette urinaire, le test à l'acide sulfosalicylique et de test à l'acide nitrique à un dosage quantitatif au rouge de pyrogallol. Au seuil de 0.50 g/L de protéines urinaires, nous avons établi que la bandelette était un très bon test de dépistage chez le chien (sensibilité de 96 à 100%) : pour un résultat de 0 ou Trace , le dépistage est négatif et positif s'il est supérieur à 2+. Si le résultat est 1+ ou 2+, il est recommandé d'effectuer un test de précipitation à l'acide sulfosalicylique ou à l'acide nitrique pour écarter les nombreux faux positifs. Les tests de précipitation par les acides ne sont pas utilisables seuls du fait de leur faible sensibilité et spécificité

Integrated barcode chips for rapid, multiplexed analysis of proteins in microliter quantities of blood

As the tissue that contains the largest representation of the human proteome [1], blood is the most important fluid for clinical diagnostics [2, 3, 4]. However, although changes of plasma protein profiles reflect physiological or pathological conditions associated with many human diseases, only a handful of plasma proteins are routinely used in clinical tests. Reasons for this include the intrinsic complexity of the plasma proteome [1], the heterogeneity of human diseases and the rapid degradation of proteins in sampled blood [5]. We report an integrated microfluidic system, the integrated blood barcode chip that can sensitively sample a large panel of protein biomarkers over broad concentration ranges and within 10 min of sample collection. It enables on-chip blood separation and rapid measurement of a panel of plasma proteins from quantities of whole blood as small as those obtained by a finger prick. Our device holds potential for inexpensive, noninvasive and informative clinical diagnoses, particularly in point-of-care settings

Non-target screening with high-resolution mass spectrometry: critical review using a collaborative trial on water analysis

In this article, a dataset from a collaborative nontarget screening trial organised by the NORMAN Association is used to review the state-of-the-art and discuss future perspectives of non-target screening using high-resolution mass spectrometry in water analysis. A total of 18 institutes from 12 European countries analysed an extract of the same water sample collected from the River Danube with either one or both of liquid and gas chromatography coupled with mass spectrometry detection. This article focuses mainly on the use of high resolution screening techniques with target, suspect, and non-target workflows to identify substances in environmental samples. Specific examples are given to emphasise major challenges including isobaric and co-eluting substances, dependence on target and suspect lists, formula assignment, the use of retention information, and the confidence of identification. Approaches and methods applicable to unit resolution data are also discussed. Although most substances were identified using high resolution data with target and suspect-screening approaches, some participants proposed tentative non-target identifications. This comprehensive dataset revealed that nontarget analytical techniques are already substantially harmonised between the participants, but the data processing remains time-consuming. Although the objective of a Bfullyautomated identification workflow^ remains elusive in the short term, important steps in this direction have been taken, exemplified by the growing popularity of suspect screening approaches. Major recommendations to improve non-target screening include better integration and connection of desired features into software packages, the exchange of target and suspect lists, and the contribution of more spectra from standard substances into (openly accessible) databases.This work was supported in part by the SOLUTIONS project, which received funding from the European Union’s Seventh Framework Programme for research, technological development and demonstration under Grant Agreement No. 603437

Comparaison des tests de dépistage rapide et de la mesure de la protéinurie chez le chien

Les tests de dépistage des protéinuries utilisables par le vétérinaire praticien ont été établis pour une utilisation en médecine humaine. La détermination de leur valeur diagnostique chez le chien est nécessaire car ces tests ne sont pas validés dans cette espèce. Le but de cette étude a été de comparer une bandelette urinaire, le test à l acide sulfosalicylique et de test à l acide nitrique à un dosage quantitatif au rouge de pyrogallol. Au seuil de 0.50 g/L de protéines urinaires, nous avons établi que la bandelette était un très bon test de dépistage chez le chien (sensibilité de 96 à 100%) : pour un résultat de 0 ou Trace , le dépistage est négatif et positif s il est supérieur à 2+. Si le résultat est 1+ ou 2+, il est recommandé d effectuer un test de précipitation à l acide sulfosalicylique ou à l acide nitrique pour écarter les nombreux faux positifs. Les tests de précipitation par les acides ne sont pas utilisables seuls du fait de leur faible sensibilité et spécificité.TOULOUSE3-BU Santé-Centrale (315552105) / SudocTOULOUSE-EN Vétérinaire (315552301) / SudocSudocFranceF

Comparaison des techniques de dépistage des protéinuries du Chien

Le dépistage d’une protéinurie est nécessaire au diagnostic et au pronostic des affections rénales du chien. Il repose sur l’utilisation de bandelettes réactives ou de tests de dénaturation par les acides nitrique ou sulfosalicylique qui n’ont pas été validés chez le Chien. Cette étude a donc évalué leur efficacité dans 151 urines canines par comparaison au dosage quantitatif des protéines totales dans l’urine par le rouge de pyrogallol. Pour des concentrations protéiques urinaires comprises entre 0,06 g/L et 13,4 g/L, il a été montré que : 1/ les bandelettes dépistaient toutes les protéinuries supérieures à 0,3 g/L ou 0,5 g/L mais donnaient un pourcentage élevé de faux positifs ; 2/ le test à l’acide sulfosalicylique donnait des résultats très dispersés ; 3/ la réaction de Heller à l’acide nitrique ne donnait que très peu de faux positifs au seuil de 0,5 g/L. En pratique, le caractère permanent d’une protéinurie devant être démontré par la répétition des analyses au moins trois fois à au moins deux semaines d’intervalle, les bandelettes sont des moyens suffisants avant d’aborder la quantification de la protéinurie et/ou la détermination du rapport protéines/créatinine urinaire

Unravelling Light and Microbial Activity as Drivers of Organic Matter Transformations in Tropical Headwater Rivers

Connecting tropical rainforests to larger rivers, tropical headwaters export large quantities of carbon and nutrients as dissolved organic matter (DOM), and are thus a key component of the global carbon cycle. This DOM transport is not passive, however; sunlight and microbial activity alter DOM concentrations and compositions, affecting riverine greenhouse gas emissions and downstream ecosystems. The effects of sunlight and microbial turnover/activity on DOM concentrations and compositions in tropical headwaters are currently poorly understood, but novel Size Exclusion Chromatography (SEC) techniques coupled to suitable detectors can for the first time quantify their influences. Here, we present in-situ incubation experiments from from headwaters of the Essequibo River, in the Iwokrama Rainforest, Guyana, where sunlight oxidised up to 9% of dissolved organic carbon (DOC) over 12 hours, at higher rates than in larger tropical rivers. DOM transformations occurred in both photo-sensitive and supposedly photo-resistant pools. Microbial activity had varying, less clear influences on DOC concentrations over the same time span; compositionally, this appeared to extend beyond known bio-labile components. Biopolymers were particularly reactive to both processes. We show sunlight has the greater potential to mineralise headwater DOM and thus potentially influence degassing. Our approach provides a future template to constrain DOM transformations along river networks, identify biogeochemical activity hotspots, and improve greenhouse gas emissions estimations under changing environmental conditions.</p

Transcriptome in paraffin samples for the diagnosis and prognosis of adrenocortical carcinoma

DESIGN: Molecular classification is important for the diagnosis and prognosis of adrenocortical tumors (ACT). Transcriptome profiles separate adrenocortical adenomas ‘C2’ from carcinomas, and identify two groups of carcinomas ‘C1A’ and ‘C1B’, of poor and better prognosis respectively. However, many ACT cannot be profiled because of improper or absent freezing procedures, a mandatory requirement so far. The main aim was to determine transcriptome profiles on formalin-fixed paraffin-embedded (FFPE) samples, using the new 3’-end RNA-sequencing technology. A secondary aim was to demonstrate the ability of this technique to explore large FFPE archives, by focusing on the rare oncocytic ACT variants. METHODS: We included 131 ACT: a training cohort from Cochin hospital and an independent validation cohort from Wuerzburg hospital. The 3’ transcriptome was generated from FFPE samples using QuantSeq (Lexogen, Vienna, Austria) and NextSeq500 (Illumina, San Diego, CA, USA). RESULTS: In the training cohort, unsupervised clustering identified three groups: ‘C1A’ aggressive carcinomas (n = 28, 29%), ‘C1B’ more indolent carcinomas (n = 28, 29%), and ‘C2’ adenomas (n = 39, 41%). The prognostic value of FFPE transcriptome was confirmed in the validation cohort (5-year OS: 26% in ‘C1A’ (n = 26) and 100% in ‘C1B’ (n = 10), P  = 0.003). FFPE transcriptome was an independent prognostic factor in a multivariable model including tumor stage and Ki-67 (OS HR: 7.5, P  = 0.01). Oncocytic ACT (n = 19) did not form any specific cluster. Oncocytic carcinomas (n = 6) and oncocytic ACT of uncertain malignant potential (n = 4) were all in ‘C1B’. CONCLUSIONS: The 3’ RNA-sequencing represents a convenient solution for determining ACT molecular class from FFPE samples. This technique should facilitate routine use and large retrospective studies

Non-target screening with high-resolution mass spectrometry: critical review using a collaborative trial on water analysis

In this article, a dataset from a collaborative non-target screening trial organised by the NORMAN Association is used to review the state-of-the-art and discuss future perspectives of non-target screening using high-resolution mass spectrometry in water analysis. A total of 18 institutes from 12 European countries analysed an extract of the same water sample collected from the River Danube with either one or both of liquid and gas chromatography coupled with massspectrometry detection. This article focuses mainly on the use of highresolution screening techniques with target, suspect, and non-target workflows to identify substances in environmental samples. Specific examples are given to emphasise major challenges including isobaric and co-eluting substances, dependence on target and suspect lists, formula assignment, the use of retention information, and the confidence of identification. Approaches and methods applicable to unit resolution data are also discussed. Although most substances were identified using highresolution data with target and suspect-screening approaches, some participants proposed tentative non-target identifications. This comprehensive dataset revealed that non-target analytical techniques are already substantially harmonised between the participants, but the data processing remains time-consuming. Although the objective of a "fully-automated identification workflow” remains elusive in the short term, important steps in this direction have been taken, exemplified by the growing popularity of suspectscreening approaches. Major recommendations to improve non-target screening include better integration and connection of desired features into software packages, the exchange of target and suspect lists, and the contribution of more spectra from standard substances into (openly accessible) databases. Graphical Abstract Matrix of identification approach versus identification confidenc