Assessment of change in psychoanalysis : Another way of using the change after psychotherapy scales

A systematic method is presented whereby material from a full course of psychoanalytic treatment is analyzed to assess changes and identify patterns of change. Through an analysis of session notes, changes were assessed using the CHange After Psychotherapy scales (CHAP; Sandell 1987a), which evaluate changes in five rating variables (symptoms, adaptive capacity, insight, basic conflicts, and extratherapeutic factors). Change incidents were identified in nearly every session. Early in the analysis, relatively more change incidents related to insight were found than were found for the other types of change. By contrast, in the third year and part of the fourth year, relatively more change incidents related to basic conflicts and adaptive capacity were found. While changes related to symptoms occurred throughout the course of treatment, such changes were never more frequent than other types of change. A content analysis of the change incidents allowed a determination of when in the treatment the patient's main conflicts (identified clinically) were overcome. A crossing of quantitative data with clinical and qualitative data allowed a better understanding of the patterns of change.

Development of dendritic cells in the intestine

The intestinal tract is exposed to a large variety of antigens such as food proteins, commensal bacteria and pathogens and contains one of the largest arms of the immune system. The intestinal immune system has to discriminate between harmless and harmful antigens, inducing tolerance to harmless antigens and active immunity towards pathogens and other harmful materials. Dendritic cells (DC) in the mucosal lamina propria (LP) are central to this process, as they sample bacteria from the local environment and constitutively migrate to the draining mesenteric lymph nodes (MLN), where they present antigen to naïve T cells in order to direct an appropriate immune response. Despite their crucial role, understanding the function and phenotype of LP DC has been hampered by the fact that they share phenotypic markers with macrophages (mφ), which are the dominant population of mononuclear phagocyte (MP) in the LP. Recent work in our own and other laboratories has established gating strategies and phenotyping panels that allow precise discrimination between intestinal DC and mφ using the mφ specific markers CD64 and F4/80. In this way four bona fide DC subsets with distinct functions have been identified in adult LP based on their expression of CD11b and CD103 and a major aim of my project was to understand how these subsets might develop in the neonatal intestine. At the beginning of my PhD, the laboratory had used these new methods to show that signal regulatory protein α (SIRPα), an inhibitory receptor expressed by myeloid cells, was expressed by mφ and most DC in the intestine, except for those expressing CD103 alone. In addition, mice carrying a non-signalling mutation in SIRPα (SIRPα mt) had a selective reduction in CD103+CD11b+ DC, a subset which is unique to the intestinal LP. This was the basis for the initial experiments of my project, described in Chapter 3, where I investigated if the phenotype in SIRPα mt mice was intrinsic to haematopoietic cells or not. To explore this, I generated bone marrow (BM) chimeric mice by reconstituting irradiated WT mice with SIRPα mt BM, or SIRPα mt animals with WT BM. These experiments suggested that the defect in CD103+CD11b+ DC was not replicated in DC derived from BM of SIRPα origin. However as this seemed inconsistent with other data, I considered the possibility that 18 the phenotype may have been lost with age, as the BM chimeric mice were considerably older than those used in the original studies of SIRPα function. However a comparison of DC subsets in the intestine of WT and SIRPα mt mice as they aged provided no conclusive evidence to support this idea. As these experiments did show age-dependent effects on DC subsets, in Chapter 4, I went on to investigate how the DC populations appeared in the intestine and other tissues in the neonatal period. These experiments showed there were few CD103+CD11b+ DC present in the LP and migratory DC compartment of the MLN in the neonate and that as this population gradually increased in proportion with age, there was a reciprocal decrease in the relative proportion of CD103-CD11b+ DC. Interestingly, most of the changes in DC numbers in the intestine were found during the second or third week of life when the weaning process began. To validate my findings that there were few CD103+CD11b+ DC in the neonate and that this was not merely an absence of CD103 upregulation, I examined the expression of CD101 and Trem-1, markers that other work in the laboratory had suggested were specific to the CD103+CD11b+ DC lineage. My work showed that CD101 and Trem-1 were co- expressed by most CD103+CD11b+ DC in small intestine (SI) LP, as well as a small subset of CD103-CD11b+ DC in this tissue. Interestingly, Trem-1 was highly specific to the SI LP and migratory DC in the MLN, but absent from the colon and other tissues. CD101 expression was also only found on CD11b+ DC, but showed a less restricted pattern of distribution, being found in several tissues as well as the SI LP. The relative timing of their development suggested there might be a relationship between CD103+CD11b+ and CD103-CD11b+ DC and this was supported by microarray analysis. I hypothesised that the CD103-CD11b+ DC that co-expressed CD101 and Trem-1 may be the cells that developed into CD103+CD11b+ DC. To investigate this I analysed how CD101 and Trem-1 expression changed with age amongst the DC subsets in SI LP, colonic LP (CLP) and MLN. The proportion of CD101+Trem-1+ cells increased amongst CD103+CD11b+ DC in the SI LP and MLN with age, while amongst CD103+CD11b+ DC in the CLP this decreased. This was not the same in CD103-CD11b+ DC, where CD101 and Trem-1 expression was more varied with age in all tissues. CD101 and Trem-1 were not expressed to any great extent on CD103+CD11b- or CD103-CD11b- DC. The phenotypic development of the 19 intestinal DC subsets was paralleled by the gradual upregulation of CD103 expression, while the production of retinoic acid (RA), as assessed by the AldefluorTM assay, was low early in life and did not attain adult levels until after weaning. Thus DC in the neonatal intestine take some time to acquire the adult pattern of phenotypic subsets and are functionally immature compared with their adult counterparts. In Chapter 5, I used CD101 and Trem-1 to explore the ontogeny of intestinal DC subsets in CCR2-/- and SIRPα mt mice, both of which have selective defects in one particular group of DC. The selective defect seen amongst CD103+CD11b+ DC in adult SIRPα mt mice was more profound in mice at D7 and D14 of age, indicating that it may be intrinsic to this population and not highly dependent on environmental factors that change after birth. The expression of CD101 and Trem-1 by both CD103+CD11b+ and CD103-CD11b+ DC was reduced in SIRPα mt mice, again indicating that this entire lineage was affected by the lack of SIRPα signalling. However there was also a generalised defect in the numbers of all DC subsets in many tissues from early in life, suggesting there was compromised development, recruitment or survival of DC in the absence of SIRPα signalling. In contrast to the findings in SIRPα mt mice, more CD103+CD11b+ DC co-expressed CD101 and Trem-1 in CCR2-/- mice, while there were no differences in the expression of these molecules amongst CD103-CD11b+ DC. This may suggest that CCR2+ CD103-CD11b+ DC are not the cells that express CD101 and Trem-1 that are predicted to be the direct precursors of CD103+CD11b+ DC. I also examined the expression of DC growth factor receptors on DC subsets from mice of different ages, but no clear age or subset- related patterns of the expression of mRNA for Csf2ra, Irf4, Tgfbr1 and Rara could be observed. Next, I investigated whether Trem-1 played any role in DC development. Preliminary experiments in Trem-1-/- mice show no differences between any of the DC subsets, nor were there any selective effects on individual subsets when DC development from Trem-1-/- KO and WT BM was compared in competitive chimeras. However these experiments were difficult to interpret due to viability problems and because I found an unexpected defect in the ability of Trem-1-/- BM to generate all DC, irrespective of whether they expressed Trem-1 or not. 20 The final experiments I carried out were to examine the role of the microbiota in driving the differentiation of intestinal DC subsets, based on the hypothesis that this could be one of the environmental factors that might influence events in the developing intestine. To this end I performed experiments in both antibiotic treated and germ free adult mice, both of which showed no significant phenotypic differences amongst any of the DC subsets. However the study of germ free mice was compromised by recent contamination of the colony and may not be the conclusive answer. Together the data in this thesis have shown that the population of CD103+CD11b+ DC, which is unique to the intestine, is not present at birth. These cells gradually increase in frequency over time and as this occurs there is a reciprocal decrease in the frequency of CD103-CD11b+ DC. Along with other results, this leads to the idea that there may be a linear developmental pathway from CD103-CD11b+ DC to CD103+CD11b+ DC that is driven by non-microbial factors that are located preferentially in the small intestine. My project indicates that markers such as CD101 and Trem-1 may assist the dissection of this process and highlights the importance of the neonatal period for these events.

Étude morphologique et physiologique du sang, de l'immunité naturelle et acquise chez quelques poissons Indochinois

This paper deals with a morphological study on blood of some Indochinese freshwater fishes. The study has been split in two parts: one dedicated to compare some fishes' blood and the second part focused on reaction of cells and natural immunity.

Relatório de estágio de aprofundamento de competências profissionais : intervenção psicomotora nas DID

A Psicomotricidade consiste numa intervenção por mediação corporal e expressiva que pode ser desenvolvida em vários âmbitos, nomeadamente o preventivo, o reeducativo, o terapêutico. Essa intervenção pretende a compensação da expressão motora inadequada ou inadaptada, ligada geralmente a problemas de desenvolvimento e maturação psicomotora, de comportamento, de aprendizagem e de âmbito emocional. As Dificuldades Intelectuais e Desenvolvimentais (DID) são caracterizadas pela manifestação antes dos 18 anos e por um conjunto de limitações significativas ao nível do funcionamento intelectual e do comportamento adaptativo em três domínios fundamentais: conceptual, social ou prático. O presente relatório visa abordar a importância da Psicomotricidade nas DID, através da apresentação das atividades realizadas com jovens e adultos, neste âmbito. Em síntese, no cumprimento do regulamento da unidade curricular do Mestrado em Reabilitação Psicomotora, iremos descrever as atividades desenvolvidas na Associação Quinta Essência, pormenorizando aspetos fundamentais das ações desenvolvidas, retirando conclusões sobre a repercussão formativa das ações aí tomadas, quer em termos pessoais quer profissionais.

Evaluating the effectiveness of discrete trial procedures for teaching receptive discrimination to children with autism spectrum disorders

Research has found that children with autism spectrum disorders (ASD) show significant deficits in receptive language skills (Wiesmer, Lord, & Esler, 2010). One of the primary goals of applied behavior analytic intervention is to improve the communication skills of children with autism by teaching receptive discriminations. Both receptive discriminations and receptive language entail matching spoken words with corresponding objects, symbols (e.g., pictures or words), actions, people, and so on (Green, 2001). In order to develop receptive language skills, children with autism often undergo discrimination training within the context of discrete trial training. This training entails teaching the learner how to respond differentially to different stimuli (Green, 2001). It is through discrimination training that individuals with autism learn and develop language (Lovaas, 2003). The present study compares three procedures for teaching receptive discriminations: (1) simple/conditional (Procedure A), (2) conditional only (Procedure B), and (3) conditional discrimination of two target cards (Procedure C). Six children, ranging in age from 2-years-old to 5-years-old, with an autism diagnosis were taught how to receptively discriminate nine sets of stimuli. Results suggest that the extra training steps included in the simple/conditional and conditional only procedures may not be necessary to teach children with autism how to receptively discriminate. For all participants, Procedure C appeared to be the most efficient and effective procedure for teaching young children with autism receptive discriminations. Response maintenance and generalization probes conducted one-month following the end of training indicate that even though Procedure C resulted in less training sessions overall, no one procedure resulted in better maintenance and generalization than the others. In other words, more training sessions, as evident with the simple/conditional and conditional only procedures, did not facilitate participants’ ability to accurately respond or generalize one-month following training. The present study contributes to the literature on what is the most efficient and effective way to teach receptive discrimination during discrete trial training to children with ASD. These findings are critical as research shows that receptive language skills are predictive of better outcomes and adaptive behaviors in the future. ^

Global transcriptome analysis of Lactococcus garvieae strains in response to temperature

Lactococcus garvieae is an important fish and an opportunistic human pathogen. The genomic sequences of several L. garvieae strains have been recently published, opening the possibility of global studies on the biology of this pathogen. In this study, a whole genome DNA microarray of two strains of L. garvieae was designed and validated. This DNA microarray was used to investigate the effects of growth temperature (18°C and 37°C) on the transcriptome of two clinical strains of L. garvieae that were isolated from fish (Lg8831) and from a human case of septicemia (Lg21881). The transcriptome profiles evidenced a strain-specific response to temperature, which was more evident at 18°C. Among the most significant findings, Lg8831 was found to up-regulate at 18°C several genes encoding different cold-shock and cold-induced proteins involved in an efficient adaptive response of this strain to low-temperature conditions. Another relevant result was the description, for the first time, of respiratory metabolism in L. garvieae, whose gene expression regulation was temperature-dependent in Lg21881. This study provides new insights about how environmental factors such as temperature can affect L. garvieae gene expression. These data could improve our understanding of the regulatory networks and adaptive biology of this important pathogen.

Implantation d’une innovation au sein d’une région rurale éloignée au Québec : le rôle d’infirmière praticienne spécialisée en soins de première ligne- une étude de cas multiples

Introduction : La situation de l’accès, de la continuité et de la coordination des services de santé au Canada et au Québec est des plus préoccupantes. Pour contribuer à résoudre ces problématiques, l’élargissement des champs de pratique professionnels a été proposé. Lorsqu’il est question d’élargir le rôle des infirmières, la pratique infirmière avancée (PIA) est fréquemment abordée. Au Québec, ce n’est qu’en 2006 qu’un rôle associé à la PIA en première ligne a pu officiellement être mis en place, celui d’infirmière praticienne spécialisée en soins de première ligne (IPSPL) (Durand, Allard, & Ménard, 2006). L’implantation du rôle d’IPSPL est récente et peut être conçue comme une innovation. Les difficultés liées à l’implantation des rôles de PIA font l’objet d’un consensus. Pour pallier à ces difficultés, il est proposé d’approfondir la compréhension de la façon dont les rôles de PIA sont implantés, et ce, en considérant les contextes. Encore peu de recherches s’intéressent au processus d’implantation du rôle d’IPSPL au Québec, et aucune n’est centrée sur le contexte rural éloigné. But : Cette recherche vise à comprendre le processus d’implantation du rôle d’IPSPL au sein d’une région rurale éloignée du Québec, à travers l’éclairage d’un cadre de référence intégrant les théories de la diffusion de l’innovation et des transitions. Méthode : Cette étude de trois cas se situe dans un paradigme pragmatique, avec des visées descriptive et explicative. Des stratégies de collecte de données mixtes ont été utilisées auprès de personnes provenant du contexte québécois, de la région ciblée et des cas (IPSP, médecins partenaires, DSI, DSP, gestionnaires, personnes soignées et leur famille). Résultats : L’implantation est un processus multidimensionnel, multifactoriel et évolutif. Le contexte, le déroulement, la compréhension, les acteurs et le temps sont des parties intégrantes de l’implantation et sont étroitement inter-reliés. Le déroulement de l’implantation et des transitions se produit simultanément. Discussion : Cette recherche a permis de mettre en lumière la raison pour laquelle le processus d’implantation du rôle d’IPSPL doit être considéré comme un processus complexe. Cette thèse contribue à éclairer la recherche axée sur l’efficacité en permettant de mieux comprendre les différentes composantes de l’implantation. Mots-clés : implantation, rôle, infirmière praticienne, première ligne, rural, éloigné, innovation, transition.

Identificación de las relaciones reales y posibles entre la administración basada en modelos, la ciencia computacional de la organización y la ingeniería emergente en el contexto de la gestión organizacional

Las teorías administrativas se han basado, casi sin excepción, en los fundamentos y los modelos de la ciencia clásica (particularmente, en los modelos de la física newtoniana). Sin embargo, las organizaciones actualmente se enfrentan a un mundo globalizado, plagado de información (y no necesariamente conocimiento), hiperconectado, dinámico y cargado de incertidumbre, por lo que muchas de las teorías pueden mostrar limitaciones para las organizaciones. Y quizá no por la estructura, la lógica o el alcance de las mismas, sino por la falta de criterios que justifiquen su aplicación. En muchos casos, las organizaciones siguen utilizando la intuición, las suposiciones y las verdades a medias en la toma de decisiones. Este panorama pone de manifiesto dos hechos: de un lado, la necesidad de buscar un método que permita comprender las situaciones de cada organización para apoyar la toma de decisiones. De otro lado, la necesidad de potenciar la intuición con modelos y técnicas no tradicionales (usualmente provenientes o inspiradas por la ingeniería). Este trabajo busca anticipar los pilares de un posible método que permita apoyar la toma de decisiones por medio de la simulación de modelos computacionales, utilizando las posibles interacciones entre: la administración basada en modelos, la ciencia computacional de la organización y la ingeniería emergente.

Effects of dark or of red, blue or white light on germination of subterranean clover seeds

Dry or imbibed seeds of the negatively photoblastic burr burying subterranean clover cv. ‘Seaton Park’ were treated with dark or with red, blue or white light to evaluate the effects of light on seed germination. Dry seeds treated with constant white light, red light or blue light during 8 days and subsequently incubated in dark had final germination and duration of germination reduced, and the distribution of germination changed from highly asymmetric to symmetric respectively. Imbibing seeds incubated under constant blue or white light had final germination strongly reduced seven days after sowing (7.3% and 50.1% of the germination under dark) with significant differences between them. After transferral to dark, true complete recovery of germination of seeds treated with white light was observed 19 days after sowing, but only partial recovery in seeds treated with blue light. Results of dry and imbibed seeds are consistent with no activity of phytochromes, as expected in negatively photoblastic seeds. Results of dry seeds are seemingly contradictory because total germination data imply the inactivity of red and blue light photoreceptors, the opposite being implied by duration and shape of germination. A tentative hypothetical solution for the contradiction is presented. Results of imbibed seeds are fully consistent with cryptochromes but not with phototropins mediation of responses to light of seed germination in ‘Seaton Park’. The ecological and adaptive significance of such responses are discussed in the framework of light attenuation in soil and the requirement and ability of subterranean clover ‘Seaton Park’ to bury seeds.

Specificità, efficacia e risposte adattative mediate da mutazioni dell'oncosoppressore TP53 degli inibitori del Complesso I mitocondriale nella terapia oncologica personalizzata: dai meccanismi molecolari allo sviluppo di modelli preclinici avanzati

L'inibizione del complesso respiratorio I (CI) è una strategia antitumorale emergente, sebbene la specificità e l’efficacia di nuovi farmaci restino poco investigate. La generazione di modelli cellulari tumorali nulli per il CI rivela la specificità di EVP 4593 e BAY 872243 nell’indurre gli effetti antiproliferativi non associati all’apoptosi, selettivamente via CI, riducendo eventuali effetti collaterali. Studi preliminari in vivo evidenziano un rallentamento della crescita tumorale negli animali trattati con EVP 4593, il quale emerge come l’inibitore più potente. Per il suo ruolo nella riprogrammazione metabolica, e la sua elevata frequenza di mutazioni nelle neoplasie umane, sono stati investigati i potenziali meccanismi di adattamento alla terapia anti-CI sulla base dello stato mutazionale di TP53. L’auxotrofia da aspartato, un hallmark metabolico delle cellule tumorali con un danno al CI, causa un blocco della sintesi proteica mTORC1-dipendente nelle linee cellulari con una p53 mutata o nulla, inducendo un collasso metabolico. Viceversa, l'attivazione del sensore energetico AMPK promuove un recupero parziale della sintesi di aspartato in linee cellulari con la forma wild type di P53, che è in grado di sostenere una migliore anaplerosi attraverso SCO2, fattore di assemblaggio del complesso respiratorio IV. Al fine di traslare questi risultati in un modello preclinico, si è ottimizzato l’ottenimento di colture di tumori umani espiantati tramite il bioreattore U-CUP. Il modello scelto è stato quello di carcinoma sieroso ad alto grado dell’ovaio (HGSOC), a partire da tessuto congelato, per l’elevata frequenza di mutazioni driver in TP53. I tessuti congelati preservano l'eterogeneità delle componenti cellulari del tessuto di origine e sono caratterizzati da cellule in attiva proliferazione senza attivazione di apoptosi. Dati preliminari mostrano un trend di riduzione dell’area tumorale nei tessuti trattati con EVP 4593 e supportano l’utilizzo del modello preclinico nello studio di nuovi inibitori del CI sfruttando materiale primario di pazienti oncologici.

Innate immunity and regulatory T-cells in human Chagas disease: what must be understood?

There is a general consensus that during chronic Trypanosoma cruzi infection, the host immune system induces complex processes to ensure the control of parasite growth while preserving the potential to mount and maintain a life-long controlled humoral and cellular immune response against the invading pathogen. This review summarises evidence in an attempt to elucidate "what must be understood" to further clarify the role of innate immunity in the development/maintenance of clinical Chagas disease and the impact of etiological treatment on host immunity, highlighting the contributions of the innate immunity and regulatory T (Treg) cells. Recently, increasing focus on innate immunity suggest that chronic T. cruzi infection may cause morbidity when innate effector functions, or the down-regulation of adaptive regulatory mechanisms are lacking. In this context, stable asymptomatic host-parasite interactions seem to be influenced by the effector/regulatory balance with the participation of macrophages, natural killer (NK) and CD8+ T cells in parallel with the establishment of regulatory mechanisms mediated by NKT and Treg cells. Moreover, a balanced innate immune activation state, apart from Treg cells, may play a role in controlling the adverse events triggered by the massive antigen release induced by trypanosomicidal agents during Chagas disease etiological treatment.

The long pentraxin PTX3 at the crossroads between innate immunity and tissue remodelling

Innate immunity represents the first line of defence against pathogens and plays key roles in the activation and orientation of the adaptive immune response. The innate immune system comprises both a cellular and a humoral arm. Components of the humoral arm include soluble pattern recognition molecules that recognize pathogen-associated molecular patterns and initiate the immune response in coordination with the cellular arm, therefore acting as functional ancestors of antibodies. Pentraxins are essential constituents of the humoral arm of innate immunity and represent a superfamily of highly conserved acute phase proteins, traditionally classified into short and long pentraxins. Pentraxin 3 (PTX3) is the prototypic member of the long pentraxins subfamily. As opposed to C-reactive protein, whose sequence and regulation have not been conserved during evolution from mouse to man, the evolutionary conservation of sequence, gene organization and regulation of PTX3 has allowed addressing its pathophysiological roles in genetically modified mice, in diverse conditions, ranging from infections to sterile inflammation, angiogenesis and female fertility. Despite this conservation, a number of predominantly non-coding polymorphisms have been identified in the PTX3 gene which, when associated in particular haplotypes, have been shown to be relevant in clinical conditions including infection and fertility. Here we review the studies on PTX3, with emphasis on pathogen recognition, tissue remodelling and crosstalk with other components of the innate immune system.

Direct effect of Plasmodium vivax recombinant vaccine candidates AMA-1 and MSP-1(19) on the innate immune response

The recombinant apical membrane antigen 1 (AMA-1) and 19-kDa fragment of merozoite surface protein (MSP-1(19)) are the lead candidates for inclusion in a vaccine against blood stages of malaria due to encouraging protective studies in humans and animals. Despite the importance of an efficacious malaria vaccine, vaccine-related research has focused on identifying antigens that result in protective immunity rather than determining the nature of anti-malarial immune effector mechanisms. Moreover, emphasis has been placed on adaptive rather than innate immune responses. In this study, we investigated the effect of Plasmodium vivax vaccine candidates Pv-AMA-1 and Pv-MSP-1(19) on the immune response of malaria-naive donors. Maturation of dendritic cells is altered by Pv-AMA-1 but not Pv-MSP-1(19), as observed by differential expression of cell surface markers. In addition, Pv-AMA-1 induced an increased production of MIP-1 alpha/CCL3 and decreased production of TARC/CCL17 levels in both dendritic cells (DCs) and peripheral blood mononuclear cells (PBMCs). Finally, a significant pro-inflammatory response was elicited by Pv-AMA-1-stimulated PBMCs. These results suggest that the recombinant vaccine candidate Pv-AMA-1 may play a direct role on innate immune response and might be involved in parasite destruction. (C) 2007 Elsevier Ltd. All rights reserved.

The anti-tumor activity of IL-12: Mechanisms of innate immunity that are model and dose dependent

IL-12 has been demonstrated to have potent anti-tumor activities in a variety of mouse tumor models, but the relative roles of NK, NKT, and T cells and their effector mechanisms in these responses have not been fully addressed. Using a spectrum of gene-targeted or Ab-treated mice we have shown that for any particular tumor model the effector mechanisms downstream of IL-12 often mimic the natural immune response to that tumor. For example, metastasis of the MHC class I-deficient lymphoma, EL4-S3, was strictly controlled by NK cells using perforin either naturally or following therapy with high-dose IL-12. Intriguingly, in B16F10 and RM-1 tumor models both NK and NKT cells contribute to natural protection from tumor metastasis, In these models, a lower dose of IL-12 or delayed administration of IL-12 dictated a greater relative role of NKT cells in immune protection from tumor metastasis. Overall, both NK and NKT cells can contribute to natural and IL-12-induced immunity against tumors, and the relative role of each population is turner and therapy dependent.