Développement d’une méthode de transfert de protéines présentes dans des sections tissulaires minces sur des cibles fonctionnalisées pour augmenter la spécificité de l’imagerie MS du protéome

L’imagerie par spectrométrie de masse (IMS) est une technique en pleine expansion et utilisée dans beaucoup d’études effectuées sur des systèmes biologiques tels que la corrélation entre l’expression moléculaire et l’état de santé d’un tissu et pour étudier la biologie du développement. Cependant, plus particulièrement lors de l’analyse de protéines, seulement les molécules les plus abondantes et/ou les plus facilement ionisables seront détectées. L’une des approches utilisées pour éviter cette limitation est de transférer les protéines de manière sélective à partir d’une section tissulaire mince vers une surface fonctionnalisée tout en maintenant leur organisation spatiale. Dans ce cas, seulement les protéines possédant une affinité pour la surface seront alors retenues alors que les autres seront éliminées. Donc, la nature chimique de cette surface est critique. Les travaux de recherches présentés dans ce mémoire portent sur le développement d’une méthode de transfert des protéines d’une section tissulaire vers une surface composée de nitrocellulose. Cette méthode utilise un système permettant d’effectuer le transfert sans contact physique direct entre les surfaces. De plus, lors du transfert, une pression physique est appliquée. Dans une première approche, la méthode développée a été appliquée en utilisant une section de rein de souris comme échantillon modèle. Des sections sérielles ont été collectées, soit pour être colorées à l’aide d’hématoxyline et d’éosine (H&E) afin de démontrer la régiospécificité du transfert, soit pour être analysées directement par IMS afin de déterminer si les protéines détectées après transfert sont également détecter dans les sections analysées directement. Les résultats obtenus ont démontré qu’un sous-ensemble de protéines a été transféré tout en conservant leur position spatiale initiale dans les sections. Certains signaux observés pour les protéines transférées sont uniques et/ou sont nettement mieux détectés que lors de l’analyse directe d’une section.

Stabilité de la relation d'activation père-enfant entre 12-18 mois et 3-5 ans et les facteurs associés à l'instabilité

La présente étude longitudinale a pour objectif d’évaluer la stabilité de la relation d’activation père-enfant entre 12-18 mois et 3-5 ans. De plus, nous nous intéressons aux facteurs qui pourraient être associés à la stabilité ou l’instabilité. La situation risquée a été conduite auprès des 39 dyades père-enfant aux deux temps de mesures, soit lorsque l’enfant était âgé entre 12 et 18 mois et lorsqu’il était âgé entre 3 et 5 ans. Le questionnaire d’ouverture au monde ainsi que le questionnaire d’évènements de vie ont été passés aux pères. Les résultats démontrent que seulement 46 % des enfants sont demeurés stables. L’instabilité et la stabilité ne se sont pas révélées associées significativement à aucun des facteurs mesurés.

La mort de mon père

Travail créatif / Creative Work

L’impact des principes éthiques sur le marché d’assurance en présence de tests génétiques

Full Text / Article complet

Perspective maternelle de la consultation anténatale pour travail préterme : une approche relationnelle

Une femme à risque d’un accouchement prématuré vit un enjeu de santé très éprouvant et stressant ; elle sera souvent hospitalisée pour recevoir des traitements médicaux visant à prolonger la grossesse et améliorer le pronostic du bébé. Dans ce contexte, une consultation avec un néonatalogiste est demandée. Plusieurs associations professionnelles médicales ont émis des lignes directrices sur cette consultation, insistant sur le besoin d’informer les parents au sujet des complications potentielles de la prématurité pour leur enfant. Ces recommandations s’inspirent du point de vue médical, et très peu d’études ont examiné la perspective, les attentes et les besoins des parents à risque d’un accouchement prématuré. Ce projet de thèse a pour objectif de proposer un modèle de relation médecin-patient informé de la perspective maternelle de la consultation anténatale, pour développer une approche clinique répondant à leurs besoins. Afin d’examiner cette problématique de façon complète, un travail constant de va-et-vient a été effectué entre la recension de données empiriques et une réflexion normative bioéthique féministe. Un projet de recherche empirique a d’abord permis d’explorer les attentes et le vécu de ces femmes. Les participantes espéraient recevoir plus que de l’information sur les complications de la prématurité. Elles souhaitaient que le néonatologiste soit attentif à leur situation particulière et qu’il développe une relation de confiance avec elles, leur permettant d’explorer leurs futurs rôles de mères et les encourageant à formuler leurs propres questions. Le cadre théorique féministe d’autonomie relationnelle a ensuite permis de proposer une approche de soin qui sache répondre aux besoins identifiés par les patientes, tout en adressant des enjeux de pouvoir intrinsèques à la clinique, qui influencent la santé et l’autonomie de ces femmes. Cette approche insiste sur l’importance de la relation de soin en clinique, contrastant avec un modèle encourageant une vision réductrice de l’autonomie, dans laquelle un simple transfert de données scientifiques serait équivalent au respect de la norme médicolégale du consentement éclairé. Ce modèle relationnel propose des actions concrètes et pratiques, encourageant les cliniciens à entrer en relation avec chaque patiente et à considérer l’influence qu’ils exercent sur l’autonomie relationnelle de leurs patientes.

La pintura d'història: pre-rafaelistes i exòtics

Materials didàctics del grup d'investigació Observatori sobre la Didàctica de les Arts (ODAS)

Penaeus monodon larvae can be protected from Vibrio harveyi infection by pre-emptive treatment of a rearing system with antagonistic or non-antagonistic bacterial probiotics

This study shows that the disease resistance and survival rate of Penaeus monodon in a larval rearing systems can be enhanced by supplementing with antagonistic or non-antagonistic probiotics. The antagonistic mode of action of Pseudomonas MCCB 102 and MCCB 103 against vibrios was demonstrated in larval mesocosm with cultures having su⁄cient concentration of antagonistic compounds in their culture supernatant. Investigations on the antagonistic properties of Bacillus MCCB 101, Pseudomonas MCCB 102 and MCCB 103 and Arthrobacter MCCB 104 against Vibrio harveyi MCCB111under in vitro conditions revealed that Pseudomonas MCCB 102 and MCCB 103 were inhibitory to the pathogen.These inhibitory propertieswere further con¢rmed in the larval rearing systems of P. monodon. All these four probionts signi¢cantly improved larval survival in long-term treatments as well as when challengedwith a pathogenic strain ofV. harveyiMCCB111. We could demonstrate that Pseudomonas MCCB 102 andMCCB103 accorded disease resistance and a higher survival rate in P. monodon larval rearing systems throughactive antagonism of vibrios,whereas Bacillus MCCB 101 and Arthrobacter MCCB 104 functioned as probiotics through immunostimulatory and digestive enzyme-supporting modes of action.

Adrenergic and muscarinic receptor subtypes functional regulation during diabetogenesis: Effect of curcumin and vitamin D3 pre-treatment

In the present study, the initial phase was directed to confirm the effects of curcumin and vitamin D3 in preventing or delaying diabetes onset by studying the blood glucose and insulin levels in the pre-treated and diabetic groups. Behavioural studies were conducted to evaluate the cognitive and motor function in experimental rats. The major focus of the study was to understand the cellular and neuronal mechanisms that ensure the prophylactic capability of curcumin and vitamin D3. To elucidate the mechanisms involved in conferring the antidiabetogenesis effect, we examined the DNA and protein profiles using radioactive incorporation studies for DNA synthesis, DNA methylation and protein synthesis. Furthermore the gene expression studies of Akt-1, Pax, Pdx-1, Neuro D1, insulin like growth factor-1 and NF-κB were done to monitor pancreatic beta cell proliferation and differentiation. The antioxidant and antiapoptotic actions of curcumin and vitamin D3 were examined by studying the expression of antioxidant enzymes - SOD and GPx, and apoptotic mediators like Bax, caspase 3, caspase 8 and TNF-α. In order to understand the signalling pathways involved in curcumin and vitamin D3 action, the second messengers, cAMP, cGMP and IP3 were studied along with the expression of vitamin D receptor in the pancreas. The neuronal regulation of pancreatic beta cell maintenance, proliferation and insulin release was studied by assessing the adrenergic and muscarinic receptor functional regulation in the pancreas, brain stem, hippocampus and hypothalamus. The receptor number and binding affinity of total muscarinic, muscarinic M1, muscarinic M3, total adrenergic, α adrenergic and β adrenergic receptor subtypes were studied in pancreas, brain stem and hippocampus of experimental rats. The mRNA expression of muscarinic and adrenergic receptor subtypes were determined using Real Time PCR. Immunohistochemistry studies using confocal microscope were carried out to confirm receptor density and gene expression results. Cell signalling alterations in the pancreas and brain regions associated with diabetogenesis and antidiabetogenesis were assessed by examining the gene expression profiles of vitamin D receptor, CREB, phospholipase C, insulin receptor and GLUT. This study will establish the anti-diabetogenesis activity of curcumin and vitamin D3 pre-treatment and will attempt to understand the cellular, molecular and neuronal control mechanism in the onset of diabetes.Administration of MLD-STZ to curcumin and vitamin D3 pre-treated rats induced only an incidental prediabetic condition. Curcumin and vitamin D3 pretreated groups injected with MLD-STZ exhibited improved circulating insulin levels and behavioural responses when compared to MLD-STZ induced diabetic group. Activation of beta cell compensatory response induces an increase in pancreatic insulin output and beta cell mass expansion in the pre-treated group. Cell signalling proteins that regulate pancreatic beta cell survival, insulin release, proliferation and differentiation showed a significant increase in curcumin and vitamin D3 pre-treated rats. Marked decline in α2 adrenergic receptor function in pancreas helps to relent sympathetic inhibition of insulin release. Neuronal stimulation of hyperglycemia induced beta cell compensatory response is mediated by escalated signalling through β adrenergic, muscarinic M1 and M3 receptors. Pre-treatment mediated functional regulation of adrenergic and cholinergic receptors, key cell signalling proteins and second messengers improves pancreatic glucose sensing, insulin gene expression, insulin secretion, cell survival and beta cell mass expansion in pancreas. Curcumin and vitamin D3 pre-treatment induced modulation of adrenergic and cholinergic signalling in brain stem, hippocampus and hypothalamus promotes insulin secretion, beta cell compensatory response, insulin sensitivity and energy balance to resist diabetogenesis. Pre-treatment improved second messenger levels and the gene expression of intracellular signalling molecules in brain stem, hippocampus and hypothalamus, to retain a functional neuronal response to hyperglycemia. Curcumin and vitamin D3 protect pancreas and brain regions from oxidative stress by their indigenous antioxidant properties and by their ability to stimulate cellular free radical defence system. The present study demonstrates the role of adrenergic and muscarinic receptor subtypes functional regulation in curcumin and vitamin D3 mediated anti-diabetogenesis. This will have immense clinical significance in developing effective strategies to delay or prevent the onset of diabetes.

Effect of within-litter birth weight variation on piglet survival and pre-weaning weight gain in a commercial herd

There are several factors that affect piglet survival and this has a bearing on sow productivity. Ten variables that influence pre-weaning vitality were analysed using records from the Pig Industry Board, Zimbabwe. These included individual piglet birth weight, piglet origin (nursed in original litter or fostered), sex, relative birth weight expressed as standard deviation units, sow parity, total number of piglets born, year and month of farrowing, within-litter variability and the presence of stillborn or mummified littermates. The main factors that influenced piglet mortality were fostering, parity and within-litter variability especially the weight of the individual piglet relative to the average of the litter (P<0.05). Presence of a mummified or stillborn littermate, which could be a proxy for unfavourable uterine environment or trauma during the birth process, did not influence pre-weaning mortality. Variability within a litter and the deviation of the weight of an individual piglet from the litter mean, influenced survival to weaning. It is, therefore, advisable for breeders to include uniformity within the litter as a selection criterion. The recording of various variables by farmers seems to be a useful management practice to identify piglets at risk so as to establish palliative measures. Further, farmers should know which litters and which piglets within a litter are at risk and require more attention.

Influence of pre-plant densities of Meloidogyne incognita on growth and root infestation of spinach (Spinacia oleracea L.) (Amaranthaceae) – an important dimension towards enhancing crop production

Vegetables represent a main source of micro-nutrients which can improve the health status of malnourished poor in the world. Spinach (Spinacia oleracea L.) is a popular leafy vegetable in many countries which is rich with several important micro-nutrients. Thus, consuming Spinach helps to overcome micro-nutrient deficiencies. Pests and pathogens act as major yield constraints in food production. Root-knot nematodes, Meloidogyne species, constitute a large group of highly destructive plant pests. Spinach is found to be highly susceptible for these nematode attacks. Though agricultural production has largely benefited from modern technologies and innovations, some important dimensions which can minimize the yield losses have been neglected by most of the growers. Pre-plant or initial nematode density in soil is a crucial biotic factor which is directly responsible for crop losses. Hence, information on preplant nematode densities and the corresponding damage is of vital importance to develop successful control procedures to enhance crop production. In the present study, effect of seven initial densities of M. incognita, i.e., 156, 312, 625, 1250, 2,500, 5,000 and 10,000 infective juveniles (IJs)/plant (equivalent to 1000cm3 soil) on the growth and root infestation on potted spinach plants was determined in a screen house. In order to ensure a high accuracy, root infestation was ascertained by the number of galls formed, the percentage galled-length of feeder roots and galled-feeder roots, and egg production, per plant. Fifty days post-inoculation, shoot length and weight, and root length were suppressed at the lowest IJs density. However, the pathogenic effect was pronounced at the highest density at which 43%, 46% and 45% reduction in shoot length and weight, and root length, respectively, was recorded. The highest reduction in root weight (26%) was detected at the second highest density. The Number of galls and percentage galled-length of feeder roots/per plant showed significant progressive increase across the increasing IJs density with the highest mean value of 432.3 and 54%, respectively. The two shoot growth parameters and root length showed significant inverse relationship with the increasing gall formation. Moreover, the shoot and root length were shown to be mutually dependent on each other. Suppression of shoot growth of spinach greatly affects the grower’s economy. Hence, control measures are essentially needed to ensure a better production of spinach via reducing the pre-plant density below the level of 0.156 IJs/cm3.

Pre-Attentive Segmentation in the Primary Visual Cortex

Stimuli outside classical receptive fields have been shown to exert significant influence over the activities of neurons in primary visual cortexWe propose that contextual influences are used for pre-attentive visual segmentation, in a new framework called segmentation without classification. This means that segmentation of an image into regions occurs without classification of features within a region or comparison of features between regions. This segmentation framework is simpler than previous computational approaches, making it implementable by V1 mechanisms, though higher leve l visual mechanisms are needed to refine its output. However, it easily handles a class of segmentation problems that are tricky in conventional methods. The cortex computes global region boundaries by detecting the breakdown of homogeneity or translation invariance in the input, using local intra-cortical interactions mediated by the horizontal connections. The difference between contextual influences near and far from region boundaries makes neural activities near region boundaries higher than elsewhere, making boundaries more salient for perceptual pop-out. This proposal is implemented in a biologically based model of V1, and demonstrated using examples of texture segmentation and figure-ground segregation. The model performs segmentation in exactly the same neural circuit that solves the dual problem of the enhancement of contours, as is suggested by experimental observations. Its behavior is compared with psychophysical and physiological data on segmentation, contour enhancement, and contextual influences. We discuss the implications of segmentation without classification and the predictions of our V1 model, and relate it to other phenomena such as asymmetry in visual search.

Retargeting of pre-set regions on chromosome for high gene expression in mammalian cells

We have developed a system to hunt and reuse special gene integration sites that allow for high and stable gene expression. A vector, named pRGFP8, was constructed. The plasmid pRGFP8 contains a reporter gene, gfp2 and two extraneous DNA fragments. The gene gfp2 makes it possible to screen the high expression regions on the chromosome. The extraneous DNA fragments can help to create the unique loci on the chromosome and increase the gene targeting frequency by increasing the homology. After transfection into Chinese hamster ovary cells (CHO) cells, the linearized pRGFP8 can integrate into the chromosome of the host cells and form the unique sites. With FACS, 90 millions transfected cells were sorted and the cells with strongest GFP expression were isolated, and then 8 stable high expression GFP CHO cell lines were selected as candidates for the new host cell. Taking the unique site created by pRGFP8 on the chromosome in the new host cells as a targeting locus, the gfp2 gene was replaced with the gene of interest, human ifngamma, by transfecting the targeting plasmid pRIH-IFN. Then using FACS, the cells with the dimmest GFP fluorescence were selected. These cells showed they had strong abilities to produce the protein of interest, IFN-gamma. During the gene targeting experiment, we found there is positive correlation between the fluorescence density of the GFP CHO host cells and the specific production rate of IFN-gamma. This result shows that the strategy in our expression system is correct: the production of the interesting protein increases with the increase fluorescence of the GFP host cells. This system, the new host cell lines and the targeting vector, can be utilized for highly expressing the gene of interest. More importantly, by using FACS, we can fully screen all the transfected cells, which can reduce the chances of losing the best cells.

Pre-procés i segmentació en imatges d’ultrasons 2D i 3D de la pròstata

Aquest projecte s'ha dut a terme amb el Grup de visió per computador del departament d'Arquitectura i Tecnologia de Computadors (ATC) de la Universitat de Girona. Està enfocat a l'anàlisi d'imatges mèdiques, en concret s'analitzaran imatges de pròstata en relació a desenvolupaments que s'estan realitzant en el grup de visió esmentat. Els objectius fixats per aquest projecte són desenvolupar dos mòduls de processamentm d'imatges els quals afrontaran dos blocs important en el tractament d'imatges, aquests dos mòduls seran un pre-processat d'imatges, que constarà de tres filtres i un bloc de segmentació per tal de cercar la pròstata dintre de les imatges a tractar. En el projecte es treballarà amb el llenguatge de programació C++, concretament amb unes llibreries que es denominen ITK (Insight Toolkit ) i són open source enfocades al tractament d'imatges mèdiques. A part d'aquesta eina s'utilitzaran d'altres com les Qt que és una biblioteca d'eines per crear entorns gràfics

Espai corporal i praxi constructiva grafomotora a l'edat pre-escolar

En totes les investigacions sobre I'evolució ontogenetica del nen es parteix d'una serie de pressuposits que, plantejats d'una manera explícita o no, determinen el seu enfoc i les seves conclusions teoriques. La relació existent entre desenvolupament i aprenentatge n'es un dels mes importants. Aquesta relació pot considerar-se com a unidireccional (a/ el desenvolupament es una condició previa a I'aprenentatge o b/ I'aprenentatge és desenvolupament) o be com a mútua i interdependent

Training and pre-decisional bias in a multiattribute decision task. 'Formaci??n y sesgo pre-decisional en una tarea de decisi??n multiatributiva'

Resumen tomado de la publicaci??n