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Résumé Contexte Les limites des analyses conventionnelles du sperme ont mis en évidence la nécessité de moyens supplémentaires d’évaluation de la qualité du sperme. Matériel et Méthodes La méthode d’électrophorèse unidimensionnelle a été utilisée pour évaluer la teneur et la qualité protéique dans des échantillons d’éjaculats individuels de 245 hommes avec des paramètres spermatiques connus. Résultats La teneur en protéines du sperme variait d’un échantillon à l’autre, en particulier dans la gamme des poids moléculaires élevés. L’intensité des bandes de 80–110 kDa était corrélée à la mobilité progressive (r = 0,15, p = 0,015) et était significativement plus élevée (p = 0,0367) dans le groupe des hommes avec des paramètres conventionnels spermatiques supérieurs aux valeurs de référence (OMS, 2010) comparé au groupe d’hommes dont l’éjaculat présentait au moins un paramètre spermatique (volume de sperme, concentration de spermatozoïdes, numération des spermatozoïdes, motilité progressive, formes normales et indice d’anomalies multiples) en deça de la valeur de référence. Les bandes dans la gamme de 80–110 kDa ont été identifiées en spectrométrie de masse et se sont révélées être des protéines de la gaine fibreuse du flagelle: la protéine d’ancrage A-kinase 4, la protéine d’ancrage A-kinase 3 et l’hexokinase de type 1. Conclusion La méthode d’électrophorèse unidimensionnelle constitue une méthode simple, rapide, fiable et peu coûteuse pour analyser la qualité des protéines du sperme dans les éjaculats humains individuels.

Abstract Background Brain capillary endothelial cells (BCECs) form the physiological basis of the blood-brain barrier (BBB). The barrier function is (at least in part) due to well-known proteins such as transporters, tight junctions and metabolic barrier proteins (e.g. monoamine oxidase, gamma glutamyltranspeptidase and P-glycoprotein). Our previous 2-dimensional gel proteome analysis had identified a large number of proteins and revealed the major role of dynamic cytoskeletal remodelling in the differentiation of bovine BCECs. The aim of the present study was to elaborate a reference proteome of Triton X-100-soluble species from bovine BCECs cultured in the well-established in vitro BBB model developed in our laboratory. Results A total of 215 protein spots (corresponding to 130 distinct proteins) were identified by 2-dimensional gel electrophoresis, whereas over 350 proteins were identified by a shotgun approach. We classified around 430 distinct proteins expressed by bovine BCECs. Our large-scale gene expression analysis enabled the correction of mistakes referenced into protein databases (e.g. bovine vinculin) and constitutes valuable evidence for predictions based on genome annotation. Conclusions Elaboration of a reference proteome constitutes the first step in creating a gene expression database dedicated to capillary endothelial cells displaying BBB characteristics. It improves of our knowledge of the BBB and the key proteins in cell structures, cytoskeleton organization, metabolism, detoxification and drug resistance. Moreover, our results emphasize the need for both appropriate experimental design and correct interpretation of proteome datasets.

Low birth weight is associated with an increased risk for developing type 2 diabetes and metabolic diseases. The placental capacity to supply nutrients and oxygen to the fetus represents the main determiner of fetal growth. However, few studies have investigated the effects of maternal diet on the placenta. We explored placental adaptive proteomic processes implicated in response to maternal undernutrition. Rat term placentas from 70% food-restricted (FR30) mothers were used for a proteomic screen. Placental mitochondrial functions were evaluated using molecular and functional approaches, and ATP production was measured. FR30 drastically reduced placental and fetal weights. FR30 placentas displayed 14 proteins that were differentially expressed, including several mitochondrial proteins. FR30 induced a marked increase in placental mtDNA content and changes in mitochondrial functions, including modulation of the expression of genes implicated in biogenesis and bioenergetic pathways. FR30 mitochondria showed higher oxygen consumption but failed to maintain their ATP production. Maternal undernutrition induces placental mitochondrial abnormalities. Although an increase in biogenesis and bioenergetic efficiency was noted, placental ATP level was reduced. Our data suggest that placental mitochondrial defects may be implicated in fetoplacental pathologies.

Purpose: Universal newborn screening for sickle cell diseases (SCDs) is not currently performed in many countries concerned by this public health problem. Owing to the technical and financial limitations of standard profiling methods (IEF coupled to subsequent HPLC), ethnically targeted neonatal screening is often preferred. Here, we demonstrate that MALDI-MS-based SCD newborn screening could be considered as a potential method for a strategy to universal screening because of its high throughput, cost-effectiveness, sensitivity and ability to automatically discriminate sickle haemoglobin., Experimental Design: We carried out a retrospective study of dried blood spots from 844 Guthrie cards. Four determinations of 1000 mass spectra were performed from each tested dried blood spot., Results: The MALDI-MS-based screening was highly correlated with the reference method. Only 2.3% of the samples presented a poor spectral quality., Conclusions and Clinical Relevance: Given that the overall acquisition, data reprocessing and software-assisted classification (ClinProTools™) time for processing four mass determinations (corresponding to one sample) was around 1 min, 1000 samples can be analysed per day. Rather than seeking to detect as many different haemoglobinopathies as possible, it would become possible to use MALDI-TOF-MS to screen (at a constant cost) as many samples as possible for sickle cell disease., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)