Imaging of thoracic aortic injury

International audience

Étude de la toxicité causée par le gène C9orf72 dans la Sclérose Latérale Amyotrophique

La Sclérose Latérale Amyotrophique (SLA) est une maladie neurodégénérative qui affecte les neurones moteurs. 10% des cas sont des cas familiaux et l’étude de ces familles a mené à la découverte de plusieurs gènes pouvant causer la SLA, incluant SOD1, TARDBP et FUS. L’expansion de la répétition GGGGCC dans le gène C9orf72 est, à ce jour, la cause la plus connue de SLA. L’impact de cette expansion est encore méconnu et il reste à déterminer si la toxicité est causée par un gain de fonction, une perte de fonction ou les deux. Plusieurs gènes impliqués dans la SLA sont conservés entre le nématode Caenorhabditis elegans et l’humain. C. elegans est un vers transparent fréquemment utilisé pour des études anatomiques, comportementales et génétiques. Il possède une lignée cellulaire invariable qui inclue 302 neurones. Aussi, les mécanismes de réponse au stress ainsi que les mécanismes de vieillissement sont très bien conservés entre ce nématode et l’humain. Donc, notre groupe, et plusieurs autres, ont utilisé C. elegans pour étudier plusieurs aspects de la SLA. Pour mieux comprendre la toxicité causée par l’expansion GGGGCC de C9orf72, nous avons développé deux modèles de vers pour étudier l’impact d’une perte de fonction ainsi que d’un gain de toxicité de l’ARN. Pour voir les conséquences d’une perte de fonction, nous avons étudié l’orthologue de C9orf72 dans C. elegans, alfa-1 (ALS/FTD associated gene homolog). Les vers mutants alfa-1(ok3062) développent des problèmes moteurs causant une paralysie et une dégénérescence spécifique des neurones moteurs GABAergiques. De plus, les mutants sont sensibles au stress osmotique qui provoque une dégénérescence. D’autre part, l’expression de la séquence d’ARN contenant une répétition pathogénique GGGGCC cause aussi des problèmes moteurs et de la dégénérescence affectant les neurones moteurs. Nos résultats suggèrent donc qu’un gain de toxicité de l’ARN ainsi qu’une perte de fonction de C9orf72 sont donc toxiques pour les neurones. Puisque le mouvement du vers peut être rapidement évalué en cultivant les vers dans un milieu liquide, nous avons développé un criblage de molécules pouvant affecter le mouvement des vers mutants alfa-1 en culture liquide. Plus de 4 000 composés ont été évalués et 80 ameliore la mobilité des vers alfa-1. Onze molécules ont aussi été testées dans les vers exprimant l’expansion GGGGCC et huit diminuent aussi le phénotype moteur de ces vers. Finalement, des huit molécules qui diminent la toxicité causée par la perte de fonction de C9orf72 et la toxicité de l’ARN, deux restaurent aussi l’expression anormale de plusieurs transcrits d’ARN observée dans des cellules dérivées de patient C9orf72. Avec ce projet, nous voulons identifier des molécules pouvant affecter tous les modes de toxicité de C9orf72 et possiblement ouvrir de nouvelles avenues thérapeutiques

Vector analysis of porosity evidences bone loss at the epiphysis in the BTX rat model of disuse osteoporosis

International audience

Structure-function relationship of new crotamine isoform from the Crotalus durissus cascavella

In this work we isolated a novel crotamine like protein from the Crotalus durissus cascavella venom by combination of molecular exclusion and analytical reverse phase HPLC. Its primary structure was:YKRCHKKGGHCFPKEKICLPPSSDLGKMDCRWKRK-CCKKGS GK. This protein showed a molecular mass of 4892.89 da that was determined by Matrix Assisted Laser Desorption Ionization Time-of-flight (MALDI-TOF) mass spectrometry. The approximately pI value of this protein was determined in 9.9 by two-dimensional electrophoresis. This crotamine-like protein isolated here and that named as Cro 2 produced skeletal muscle spasm and spastic paralysis in mice similarly to other crotamines like proteins. Cro 2 did not modify the insulin secretion at low glucose concentration (2.8 and 5.6 mM), but at high glucose concentration (16.7 mM) we observed an insulin secretion increasing of 2.7-3.0-fold than to control. The Na+ channel antagonist tetrodoxin (6 mM) decreased glucose and Cro 2-induced insulin secretion. These results suggested that Na+ channel are involved in the insulin secretion. In this article, we also purified some peptide fragment from the treatment of reduced and carboxymethylated Cro 2 (RC-Cro 2) with cyanogen bromide and protease V8 from Staphylococcus aureus. The isolated pancreatic beta-cells were then treated with peptides only at high glucose concentration (16.7 mM), in this condition only two peptides induced insulin secretion. The amino acid sequence homology analysis of the whole crotamine as well as the biologically-active peptide allowed determining the consensus region of the biologically-active crotamine responsible for insulin secretion was KGGHCFPKE and DCRWKWKCCKKGSG.

Vai morrer o meu menino - Paralisado, doente, isolado, exclusivo, desejado e amado: Estudo de um processo de comunicação num universo simbiótico

Dissertação de Mestrado apresentada ao Instituto Superior de Psicologia Aplicada para obtenção de grau de Mestre na especialidade de Psicologia Clínica.

Intervenção psicomotora por terras de Moçambique : crianças com paralisia cerebral e jovens com DID, uma realidade escondida

O presente documento, criado no âmbito da Unidade Curricular de Atividade de Aprofundamento de Competências Profissionais, do Mestrado em Reabilitação Psicomotora, apresenta o trabalho desenvolvido no Centro de Reabilitação Psicossocial, das Irmãs Hospitaleiras em Maputo – Moçambique. O processo de intervenção psicomotora foi desenvolvido com jovens/adultos com Dificuldades Intelectuais e Desenvolvimentais e crianças com Paralisia Cerebral ou Atraso Global do Desenvolvimento, durante 8 meses. A intervenção concretizou-se em sessões de grupo e individuais em contexto de atividade motora, terapias expressivas, desenvolvimento pessoal e social com os jovens/adultos e em contexto de promoção do desenvolvimento psicomotor, atividades psicopedagógicas e terapias expressivas com as crianças, com objetivo de promoção de bem-estar, qualidade de vida, funcionalidade e autonomia dos utentes, tendo em conta o contexto cultural, familiar e as potencialidades, dificuldades e interesses. No processo de intervenção estiveram presentes as fases próprias da prática terapêutica, observação, conhecimento e criação de relação empática com os utentes, avaliação inicial, processo de intervenção planeado, avaliação final e comparação dos resultados com avaliação inicial e reflexão sobre o mesmo. É ainda apresentado um projeto de investigação centrado no estudo das atitudes da comunidade do Bairro das Mahotas perante a população com Dificuldade Intelectual e Desenvolvimental.

Avaliação ecotoxicológica de fármacos psicotrópicos e suas possíveis interações com nanomateriais usando embriões de peixe-zebra

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Animal, 2016.

Cold acclimation of chill coma mechanisms in the locust CNS

Temperature has profound effects on the neural function and behaviour of insects. When exposed to low temperature, migratory locusts (Locusta migratoria) enter chill coma (neuromuscular paralysis) and can resume normal body functions after returning to normal temperature. Our laboratory has studied phenomena underlying environmental stress-induced comas in locusts and found that they are associated with a sudden loss of K+ homeostasis and also a temporary electrical silence in the central nervous system (CNS). However, the mechanisms underlying chill coma entry and recovery are not well understood, particularly the role of the CNS has not been determined. Here, I investigated neural function during chill coma in the locust by measuring electrical activity in the CNS. As pre-exposure to moderately low temperatures, either chronically (cold acclimation) or acutely (rapid cold hardening; RCH), has been found to improve the insect’s cold tolerance, I also determined cold acclimation and RCH protocols that will improve the locust's cold tolerance and whether these protocols affect neural shutdown during chill coma in the locust. With an implanted thermocouple in the thorax, I determined the temperature associated with a loss of responsiveness (CTmin) in intact male adult locusts. In parallel experiments, I recorded field potential (FP) in the metathoracic ganglion (MTG) in semi-intact preparations to determine the temperature that would induce neural shutdown. I found that acclimation at 10 ˚C and RCH at 4 ˚C reduced chill coma recovery time (CCRT) in intact animal preparations and RCH at 4 ˚C for 4 hours reduced the temperature at neural shutdown in semi-intact preparations. These results suggest that pre-exposure to cold can improve the locust's resistance to chill coma and support the notion that the CNS has a role in determining entry into and exit from chill coma in locusts.

Obstructive sleep apneas naturally occur in mice during REM sleep and are more prevalent in a mouse model of Down syndrome

Study Objectives. The use of mouse models in sleep apnea research is limited by the belief that central (CSA) but not obstructive sleep apneas (OSA) occur in rodents. With this study we wanted to develop a protocol to look for the presence of OSAs in wild-type mice and, then, to apply it to a mouse model of Down Syndrome (DS), a human pathology characterized by a high incidence of OSAs. Methods. Nine C57Bl/6J wild-type mice were implanted with electrodes for electroencephalography (EEG), neck electromyography (nEMG), diaphragmatic activity (DIA) and then placed in a whole-body-plethysmographic (WBP) chamber for 8h during the resting (light) phase to simultaneously record sleep and breathing activity. The concomitant analysis of WBP and DIA signals allowed the discrimination between CSA and OSA. The same protocol was then applied to 12 Ts65Dn mice (a validated model of DS) and 14 euploid controls. Results. OSAs represented about half of the apneic events recorded during rapid-eye-movement sleep (REMS) in each experimental group while almost only CSAs were found during non-REMS. Ts65Dn mice had similar rate of apneic events than euploid controls but a significantly higher occurrence of OSAs during REMS. Conclusions. We demonstrated for the first time that mice physiologically exhibit both CSAs and OSAs and that the latter are more prevalent in the Ts65Dn mouse model of DS. These findings indicate that mice can be used as a valid tool to accelerate the comprehension of the pathophysiology of all kind of sleep apnea and for the development of new therapeutical approaches to contrast these respiratory disorders.

The United Nations 'quasi-prosecutorial' investigative mechanisms: assessing a new institutional model in the international criminal justice landscape

This work aims to provide a theoretical examination of three recently created bodies of the United Nations mandated to investigate the alleged international crimes committed in Syria (IIIM), Iraq (UNITAD) and Myanmar (IIMM). Established as a compromise solution in the paralysis of international criminal jurisdictions, these essentially overlapping entities have been depicted as a ‘new generation’ of UN investigative mechanisms. While non-judicial in nature, they depart indeed from traditional commissions of inquiry in several respects due to their increased criminal or ‘quasi-prosecutorial’ character. After clarifying their legal basis and different mandating authorities, a comparative institutional analysis is thus carried out in order to ascertain whether these ‘mechanisms’ can be said to effectively represent a new institutional model. Through an in-depth assessment of their mandates, the thesis is also intended to outline both the strengths and the criticalities of these organs. Given their aim to facilitate criminal proceedings by sharing information and case files, it is suggested that more attention shall be paid to the position of the person under investigation. To this end, some proposals are made in order to enhance the mechanisms’ frameworks, especially from the angle of procedural safeguards. As a third aspect, the cooperation with judicial authorities is explored, in order to shed light on the actors involved, the relevant legal instruments and the possible obstacles, in particular from a human rights perspective. Ultimately, drawing from the detected issues, the thesis seeks to identify some lessons learned which could be taken into account in case of creation of new ad hoc investigative mechanisms or of a permanent institution of this kind.

Adattamento morfologico e variazione dell'espressione genica dopo pneumonectomia: osservazioni su modello sperimentale animale

Introduzione La pneumonectomia su modello animale potrebbe essere un’utile piattaforma di studio per approfondire i meccanismi della risposta compensatoria al danno polmonare. Scopo dello studio è determinare la presenza di variazioni morfologiche e di espressione del trascrittoma dopo pneumonectomia. Materiali e metodi Undici suini sono stati sottoposti a pneumonectomia sinistra. Sono stati eseguiti prelievi sito-specifici intraoperatori su polmone sinistro e successivamente confrontati con prelievi sito-specifici su polmone destro dopo eutanasia a 60 giorni. I prelievi degli animali con decorso regolare sono stati sottoposti a RNA-sequencing e successiva analisi computazionale per valutare il peso funzionale del singolo gene o di clusters di geni. Risultati Un animale è stato escluso per insorgenza di ernia diaframmatica. In 7/10 è stata riscontrata apertura della pleura mediastinica con parziale erniazione del polmone controlaterale e shift mediastinico. L’istologia ha mostrato dilatazione degli spazi aerei, rottura dei setti interalveolari, lieve infiammazione, assenza di fibrosi, stiramento radiale dei bronchi e riduzione del letto capillare. L’analisi di bulk RNA-sequencing ha identificato 553 geni espressi in modo differenziale (DEG)(P<0,001) tra pre e post-pneumonectomia. I primi 10 DEG up-regolati: Edn1, Areg, Havcr2, Gadd45g, Depp1, Cldn4, Atf3, Myc, Gadd45b, Socs3; i primi 10 geni down-regolati: Obscn, Cdkn2b, ENSSSCG00000015738, Prrt2, Amer1, Flrt3, Efnb2, Tox3, Znf793, Znf365. Tra i DEG è stata riscontrata una predominanza di geni specifici dei macrofagi. L’analisi di gene ontology basata su DAVID ha mostrato un significativo arricchimento della "via di segnalazione apoptotica estrinseca"(FDR q=7,60x10 -3), della via di “Risposta all’insulina”(FDR q=7,60x10 -3) ed un arricchimento di geni “Regolatori negativi del segnale DDX58/IFIH1”(FDR q=7.50x10 -4). Conclusioni Il presente studio conferma la presenza di variazioni macroscopiche e microscopiche fenotipiche dopo pneumonectomia. L’RNA sequencing e lo studio di genomica traslazionale hanno mostrato l’esistenza di geni singoli e di network di geni disregolati dopo pneumonectomia, prevalentemente in determinate popolazioni cellulari.

Adattamento alla macchina della tosse guidato dall'ecografia diaframmatica nel paziente con patologia neuromuscolare: trial clinico controllato e semi-randomizzato.

Background Le complicazioni dovute a disfunzioni dei muscoli respiratori sono tra le cause principali di morbilità e mortalità per i pazienti con malattie neuromuscolari. La fisioterapia respiratoria, nello specifico tramite l’utilizzo della macchina della tosse, assume quindi un ruolo fondamentale nella gestione della clearance bronchiale. Questo studio si pone l’obbiettivo di verificare se l’ecografia diaframmatica può rappresentare uno strumento di valutazione utile al fisioterapista nella titolazione dei corretti parametri della macchina della tosse. Metodi Lo studio è un trial clinico semi-randomizzato controllato a gruppi paralleli e monocentrico, iniziato a gennaio 2022 e concluso ad ottobre dello stesso anno. Il gruppo sperimentale, rispetto a quello di controllo, vede aggiunta l’ecografia diaframmatica all’adattamento della macchina della tosse. L’efficacia è stata valutata analizzando la reale indicazione al trattamento, l’aderenza e il numero di riacutizzazioni. Gli outcome sono stati valutati a 3 mesi dall’adattamento. Risultati 10 pazienti hanno soddisfatto i criteri di inclusione e sono stati randomizzati nel gruppo sperimentale (n=6) e nel gruppo di controllo (n=4). Il livello di significatività statistica è stato fissato pari a 0.05. I dati ottenuti dalle analisi (test χ2 e il test di Mann-Whitney) portano alla conclusione che non sono presenti differenze statisticamente significative legate al gruppo di appartenenza dei pazienti per nessuno degli outcome prefissati: indicazione al trattamento ( χ2=1.67; P=0.197), riacutizzazioni (χ2=0.08; P=0.778), aderenza (W=3; P=1). Conclusioni L’ecografia diaframmatica si è rivelata essere uno strumento affidabile e poco invasivo, utile a perfezionare il processo valutativo fisioterapico. I dati ottenuti non hanno però dimostrato la superiorità del suo impiego nell’adattamento alla macchina della tosse rispetto al gruppo di controllo. Tuttavia, sono necessari ulteriori studi per approfondirne l’efficacia.