Archives internationales de physiologie et de biochimie.

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Évaluation formative et TIC pour une meilleure intégration des apprentissages en biochimie clinique

Dans cette recherche, l’enseignante-chercheuse analyse la méthode d’enseignement, les activités d’apprentissage et les modalités d’évaluation de la portion théorique du cours de stage en biochimie clinique dans le cadre du programme Technologie d’analyses biomédicales (140.BO) offert au Collège de Rosemont. Dans les années précédant la recherche, les stagiaires avaient formulé certains problèmes dont: un contenu trop lourd et le manque d’interactivité. Ces contraintes ont obligé l’enseignante-chercheuse à se questionner sur le processus pédagogique et sur la relation d’apprentissage. De plus, le manque d’intégration des connaissances des stagiaires mis en évidence lors de la préparation de l’examen national de la SCSLM et la baisse de performance à ces mêmes examens ont été les éléments déclencheurs de la recherche. Trois approches pédagogiques ont été utilisées afin de favoriser l’intégration des apprentissages et faire progresser les étudiantes et les étudiants vers un résultat acceptable en fin de stage. Premièrement, il a fallu passer par une réorganisation de contenu. Deuxièmement, nous avons mis en place un environnement plus interactif favorisant l’évaluation formative à l’aide de la plate-forme pédagogique Moodle. Troisièmement, nous avons introduit un mode d’évaluation formative qui s’harmonise à l’évaluation sommative. Toutes ces étapes ont été effectuées afin de faire progresser l’étudiante et l’étudiant et de l’informer clairement de ses performances pour qu’elle ou qu’il en arrive à intégrer les nombreuses notions théoriques en biochimie. Il est important de préciser que c’est à partir des assises du cadre de référence de cette recherche qu’il a été possible de dégager des actions pour innover notre pratique pédagogique. C’est par l’évaluation formative s’inscrivant dans diverses modalités de régulation ou d’autorégulation de l’apprentissage, permettant l’accès à la reprise à l’aide des questionnaires interactifs de la plate-forme pédagogique Moodle que nous avons mis en place une première solution. Comme deuxième solution, l’enseignante-chercheuse a présenté à chaque étape, par sujet de révision, une évaluation formative à des fins sommatives avec la possibilité d’une deuxième tentative. Les performances qui y ont été évaluées, ont été modifiables et ont contribué à la note de fin d’étape. Le but de notre démarche était de trouver des réponses à la question de recherche: L’évaluation formative par l’utilisation des technologies améliorera-t-elle les apprentissages des étudiantes et des étudiants en biochimie à la fin du programme d’étude en Techniques d’analyses biomédicales? La conception du scénario pédagogique ainsi que la mise à l’essai du matériel servant à l’évaluation formative à travers les outils interactifs de la plate-forme Moodle, ont tous été aussi importants. Dans le cadre du choix méthodologique, les trois objectifs spécifiques de la recherche ont été respectés, c’est-à-dire: 1. Concevoir le matériel servant à l’évaluation formative et le générer sur la plate-forme informatisée; 2. Mettre en application l’utilisation des nouveaux éléments d’apprentissage et des processus d’évaluation; 3. Vérifier si l’évaluation formative influencera l’apprentissage qui aura pour but une meilleure intégration des connaissances. Par le volet qualitatif de l’étude, l’enseignante-chercheuse vise à comprendre ce qui a été formulé par les étudiantes et les étudiants relativement à une démarche d’évaluation formative, interactive et continue générée par l’usage de l’outil informatisé Moodle. Cette démarche d’apprentissage assisté par ordinateur comprenait des applications d’évaluation formative élaborées sur la plate-forme pédagogique Moodle pour que les étudiantes et les étudiants en arrivent à cibler plus facilement leurs lacunes. Ainsi, les étudiantes et les étudiants étaient en mesure d’y remédier immédiatement par la fonction régulatrice propre à l’évaluation formative à l’aide des TIC. Ce qui situe la recherche dans le pôle de l’innovation, c’est l’intégration des TIC qui a permis d’établir les modalités d’évaluation formative à des fins sommatives, une reprise en deux temps, dans une perspective d’intégration des apprentissages. Par la collecte de données, se rangeant sous le paradigme interprétatif, la grille d’appréciation et l’entrevue semi-dirigée ont permis de faire ressortir ce que les étudiantes et les étudiants ont noté comme éléments d’approbation ou de valorisation de l’enseignement et de l’apprentissage. Dans le volet quantitatif de l’étude, l’enseignante-chercheuse a analysé les résultats des examens du parcours de stage dans l’intérêt de savoir s’il y a eu amélioration des apprentissages des étudiantes et des étudiants en stage à la fin du programme. De ces étapes, l’interprétation des données recueillies a permis d’alimenter la réflexion sur les principales approches mises en place dans les cours théoriques du stage en biochimie clinique. Cette étape a contribué à supporter l’objectivation du sens produit de cette recherche. Les résultats obtenus et l’interprétation qui en résulte présente que l’ensemble des stagiaires s’accorde à dire que les tests formatifs, le processus d’évaluation formative à des fins sommatives, une reprise en deux temps et le contenu interactif sur la plate-forme Moodle, permettent de faire des liens entre les connaissances déjà acquises, d’intégrer la matière et contribue à stimuler leur motivation à fournir un effort soutenu. L’utilisation des ressources du système de gestion de l’apprentissage Moodle a permis d’établir des modes d’évaluation formative dans une perspective d’intégration des apprentissages. Ces résultats sont intéressants puisqu’il a été possible de constater les difficultés des stagiaires aux examens, de relier des questions à des objectifs moins bien réussis et pour lesquels les étudiantes et les étudiants affirment ne pas avoir assez de temps d’enseignement à propos de ces notions théoriques. L’enseignante-chercheuse a alors entrepris une démarche d’amélioration continue de sa pratique pédagogique et de l’environnement numérique d’apprentissage. Entre 2012 et 2015, elle a utilisé une grille d’évaluation et a analysé les résultats des examens lui permettant de mieux cibler les ressources à améliorer pour favoriser la réussite de ses étudiantes et de ses étudiants. Elle poursuit sa réflexion à propos des conditions d’appropriation des connaissances théoriques en biochimie et aux interventions les plus susceptibles d’assurer l’intégration des apprentissages.

The kallikrein-related peptidase family: Dysregulation and functions during cancer progression

Cancer is the second leading cause of death with 14 million new cases and 8.2 million cancer-related deaths worldwide in 2012. Despite the progress made in cancer therapies, neoplastic diseases are still a major therapeutic challenge notably because of intra- and inter-malignant tumour heterogeneity and adaptation/escape of malignant cells to/from treatment. New targeted therapies need to be developed to improve our medical arsenal and counter-act cancer progression. Human kallikrein-related peptidases (KLKs) are secreted serine peptidases which are aberrantly expressed in many cancers and have great potential in developing targeted therapies. The potential of KLKs as cancer biomarkers is well established since the demonstration of the association between KLK3/PSA (prostate specific antigen) levels and prostate cancer progression. In addition, a constantly increasing number of in vitro and in vivo studies demonstrate the functional involvement of KLKs in cancer-related processes. These peptidases are now considered key players in the regulation of cancer cell growth, migration, invasion, chemo-resistance, and importantly, in mediating interactions between cancer cells and other cell populations found in the tumour microenvironment to facilitate cancer progression. These functional roles of KLKs in a cancer context further highlight their potential in designing new anti-cancer approaches. In this review, we comprehensively review the biochemical features of KLKs, their functional roles in carcinogenesis, followed by the latest developments and the successful utility of KLK-based therapeutics in counteracting cancer progression.

Importance of non-conserved distal carboxyl terminal amino acids in two peptidases belonging to the M1 family: Thermoplasma acidophilum Tricorn interacting factor F2 and Escherichia coli Peptidase N

Enzymes belonging to the M1 family play important cellular roles and the key amino acids (aa) in the catalytic domain are conserved. However, C-terminal domain aa are highly variable and demonstrate distinct differences in organization. To address a functional role for the C-terminal domain, progressive deletions were generated in Tricorn interacting factor F2 from Thermoplasma acidophilum (F2) and Peptidase N from Escherichia coli (PepN). Catalytic activity was partially reduced in PepN lacking 4 C-terminal residues (PepNΔC4) whereas it was greatly reduced in F2 lacking 10 C-terminal residues (F2ΔC10) or PepN lacking eleven C-terminal residues (PepNΔC11). Notably, expression of PepNΔC4, but not PepNΔC11, in E. coliΔpepN increased its ability to resist nutritional and high temperature stress, demonstrating physiological significance. Purified C-terminal deleted proteins demonstrated greater sensitivity to trypsin and bound stronger to 8-amino 1-napthalene sulphonic acid (ANS), revealing greater numbers of surface exposed hydrophobic aa. Also, F2 or PepN containing large aa deletions in the C-termini, but not smaller deletions, were present in high amounts in the insoluble fraction of cell extracts probably due to reduced protein solubility. Modeling studies, using the crystal structure of E. coli PepN, demonstrated increase in hydrophobic surface area and change in accessibility of several aa from buried to exposed upon deletion of C-terminal aa. Together, these studies revealed that non-conserved distal C-terminal aa repress the surface exposure of apolar aa, enhance protein solubility, and catalytic activity in two soluble and distinct members of the M1 family.

Homologous pairing between nucleosome cores on a linear duplex DNA and nucleoprotein filaments of RecA protein-single stranded DNA

The ability of E coli recA protein to promote homologous pairing with linear duplex DNA bound to HU protein (Nucleosome cores) was found to be differentially affected. The formation of paranemic joint molecules was not affected whereas the formation of plectomic joint molecules was inhibited from the start of the reaction. The formation of paranemic joint molecules between nucleoprotein filaments of recA protein-circular single stranded DNA and closed circular duplex DNA is believed to generate positive supercoiling in the duplex DNA. We found that the positively superhelical duplex DNA was inert in the formation of joint molecules but could be converted into an active substrate, in situ, by the action of wheat germ topoisomerase I. These observations initiate an understanding of the structural features of E coli chromosome such as DNA supercoiling and nucleosome-like structures in homologous recombination.

Effect of thiocarbamate, urea, and uracil herbicides on lipid metabolism in groundnut (Arachis hypogaea) leaves

The effect of thiocarbamates (S-ethyldipropylthiocarbamate and diallate), substituted ureas (monuron and diuron), and uracils (bromacil and terbacil) on lipid metabolism in groundnut (Arachis hypogaea) leaves was investigated under nonphotosynthetic conditions. The uptake of [1-14C]acetate by leaf disks was inhibited by the thiocarbamates and marginally by the substituted ureas, but not by the uracil herbicides. The uptake of [methyl-14C]choline was inhibited to a lesser extent by thiocarbamates, while the other herbicides showed a slight stimulation. The thiocarbamates almost completely inhibited uptake of [32P]orthophosphate at 1.0 mM concentration, while diuron and terbacil showed significant inhibition. [1-14C]Acetate incorporation into lipids was inhibited only by diallate. [methyl-14C]Choline incorporation into the choline phosphoglycerides was inhibited by diallate, diuron, and bromacil. The incorporation of [32P]orthophosphate into phospholipids was substantially inhibited (over 90% at 1.0 mM) by the thiocarbamates, but not by the other herbicides. [35S]Sulfate incorporation into sulfoquinovosyl diglycerides was markedly inhibited only by the thiocarbamates. Fatty acid synthesis by isolated chloroplasts was inhibited 40–85% by thiocarbamates, substituted ureas, and bromacil, but not by terbacil. The inhibitory effect of the urea derivatives was reversible, but that of thiocarbamates was irreversible. sn-Glycerol-3-phosphate acyltransferase(s) of the chloroplast and microsomal fractions were profoundly inhibited by thiocarbamates, but not by the other two groups of herbicides. Phosphatidic acid phosphatase was insensitive to all the herbicides tested.

Improvement of protein structure comparison using a structural alphabet

The three dimensional structure of a protein provides major insights into its function. Protein structure comparison has implications in functional and evolutionary studies. A structural alphabet (SA) is a library of local protein structure prototypes that can abstract every part of protein main chain conformation. Protein Blocks (PBS) is a widely used SA, composed of 16 prototypes, each representing a pentapeptide backbone conformation defined in terms of dihedral angles. Through this description, the 3D structural information can be translated into a 1D sequence of PBs. In a previous study, we have used this approach to compare protein structures encoded in terms of PBs. A classical sequence alignment procedure based on dynamic programming was used, with a dedicated PB Substitution Matrix (SM). PB-based pairwise structural alignment method gave an excellent performance, when compared to other established methods for mining. In this study, we have (i) refined the SMs and (ii) improved the Protein Block Alignment methodology (named as iPBA). The SM was normalized in regards to sequence and structural similarity. Alignment of protein structures often involves similar structural regions separated by dissimilar stretches. A dynamic programming algorithm that weighs these local similar stretches has been designed. Amino acid substitutions scores were also coupled linearly with the PB substitutions. iPBA improves (i) the mining efficiency rate by 6.8% and (ii) more than 82% of the alignments have a better quality. A higher efficiency in aligning multi-domain proteins could be also demonstrated. The quality of alignment is better than DALI and MUSTANG in 81.3% of the cases. Thus our study has resulted in an impressive improvement in the quality of protein structural alignment. (C) 2011 Elsevier Masson SAS. All rights reserved.

Progressive structure-based alignment of homologous proteins: Adopting sequence comparison strategies

Comparison of multiple protein structures has a broad range of applications in the analysis of protein structure, function and evolution. Multiple structure alignment tools (MSTAs) are necessary to obtain a simultaneous comparison of a family of related folds. In this study, we have developed a method for multiple structure comparison largely based on sequence alignment techniques. A widely used Structural Alphabet named Protein Blocks (PBs) was used to transform the information on 3D protein backbone conformation as a ID sequence string. A progressive alignment strategy similar to CLUSTALW was adopted for multiple PB sequence alignment (mulPBA). Highly similar stretches identified by the pairwise alignments are given higher weights during the alignment. The residue equivalences from PB based alignments are used to obtain a three dimensional fit of the structures followed by an iterative refinement of the structural superposition. Systematic comparisons using benchmark datasets of MSTAs underlines that the alignment quality is better than MULTIPROT, MUSTANG and the alignments in HOMSTRAD, in more than 85% of the cases. Comparison with other rigid-body and flexible MSTAs also indicate that mulPBA alignments are superior to most of the rigid-body MSTAs and highly comparable to the flexible alignment methods. (C) 2012 Elsevier Masson SAS. All rights reserved.

Inmigración: indicadores de nutrición

[ES] Los movimientos migratorios forman parte de la historia universal. La población inmigrante, igual que el resto de la población, puede sufrir malnutrición. Los indicadores de la nutrición en inmigrantes son los mismos que en los autóctonos. La valoración nutricional debe realizarse de forma sistematica, mediante antropometría, historia clínica y dietética, etc. Se puede realizar de forma objetiva o subjetiva.

Valoración nutricional en pacientes quirúrgicos

[ES] Numerosos estudios han demostrado que muchos de los pacientes que ingresan en Cirugía presentan signos de malnutrición. En el presente trabajo se estudió re t rospectivamente la valoración nutricional de 116 pacientes. Los resultados muestran que sólo en el 29,3% de los enfermos se determinaron parámetros antropométricos, en el 19% parámetros bioquímicos y en el 25% parámetros inmunológicos de interés nutricional. Cabe destacar que, como también indican otros autores, la valoración nutricional no constituye todavía un procedimiento habitual en la práctica clínica.

The NRF-1/α-PAL transcription factor regulates human E2F6 promoter activity

E2F6 is widely expressed in human tissues and cell lines. Recent studies have demonstrated its involvement in developmental patterning and in the regulation of various genes implicated in chromatin remodelling. Despite a growing number of studies, nothing is really known concerning the E2F6 expression regulation. To understand how cells control E2F6 expression, we analysed the activity of the previously cloned promoter region of the human E2F6 gene. DNase I footprinting, gel electrophoretic-mobility shift, transient transfection and site-directed mutagenesis experiments allowed the identification of two functional NRF-1/α-PAL (nuclear respiratory factor-1/α-palindrome-binding protein)-binding sites within the human E2F6 core promoter region, which are conserved in the mouse and rat E2F6 promoter region. Moreover, ChIP (chromatin immunoprecipitation) analysis demonstrated that overexpressed NRF-1/α-PAL is associated in vivo with the E2F6 promoter. Furthermore, overexpression of full-length NRF-1/α-PAL enhanced E2F6 promoter activity, whereas expression of its dominant-negative form reduced the promoter activity. Our results indicate that NRF-1/α-PAL is implicated in the regulation of basal E2F6 gene expression.

ERM transactivation is up-regulated by the repression of DNA binding after the PKA phosphorylation of a consensus site at the edge of the ETS domain

The final step of the transduction pathway is the activation of gene transcription, which is driven by kinase cascades leading to changes in the activity of many transcription factors. Among these latter, PEA3/E1AF, ER81/ETV1, and ERM, members of the well conserved PEA3 group from the Ets family are involved in these processes. We show here that protein kinase A (PKA) increases the transcriptional activity of human ERM and human ETV1, through a Ser residue situated at the edge of the ETS DNA-binding domain. PKA phosphorylation does not directly affect the ERM transactivation domains but does affect DNA binding activity. Unphosphorylated wild-type ERM bound DNA avidly, whereas after PKA phosphorylation it did so very weakly. Interestingly, S367A mutation significantly reduced the ERM-mediated transcription in the presence of the kinase, and the DNA binding of this mutant, although similar to that of unphosphorylated wild-type protein, was insensitive to PKA treatment. Mutations, which may mimic a phosphorylated serine, converted ERM from an efficient DNA-binding protein to a poor DNA binding one, with inefficiency of PKA phosphorylation. The present data clearly demonstrate a close correlation between the capacity of PKA to increase the transactivation of ERM and the drastic down-regulation of the binding of the ETS domain to the targeted DNA. What we thus demonstrate here is a relatively rare transcription activation mechanism through a decrease in DNA binding, probably by the shift of a non-active form of an Ets protein to a PKA-phosphorylated active one, which should be in a conformation permitting a transactivation domain to be active.