Ο Σακχαρώδης Διαβήτης τύπου 1 (ΣΔ1) αποτελεί τη συχνότερη μορφή διαβήτη στα παιδιά, συχνά συνοδεύεται από σοβαρές μικρο- και μακρο-αγγειακές επιπλοκές ενώ, η επίπτωσή του εμφανίζει ανησυχητικά αυξητική τάση τις τελευταίες δεκαετίες. Η ενεργοποίηση των μονοκυττάρων, η χημειοταξία τους (κύριος χημειοτακτικός παράγοντας μονοκυττάρων: MCP-1) και η μετατροπή τους σε μακροφάγα (δείκτης μακροφάγων: CD68) θεωρείται πως διαδραματίζουν προεξέχοντα ρόλο στην ανάπτυξη των διαβητικών αγγειακών αλλοιώσεων. Η πλεονάζουσα γλυκόζη που υπάρχει στο διαβήτη, αλληλεπιδρώντας μη ενζυματικά με πλήθος υποστρωμάτων, προκαλεί την παραγωγή των AGEs (τελικά προϊόντα προχωρημένης γλυκοζυλίωσης) τα οποία με τη σειρά τους ασκούν τις βλαπτικές τους δράσεις, είτε αλληλεπιδρώντας άμεσα με τα μόρια της εξωκυττάριας ουσίας ή μέσω σύνδεσης στους κυτταρικούς τους υποδοχείς (κυριότερος εκπρόσωπος: RAGE). Η ενεργοποίηση του RAGE, διαμέσου της ενεργοποίησης του μεταγωγικού μονοπατιού της πρωτεϊνικής κινάσης Β (Akt2/PKB) και του μεταγραφικού παράγοντα NF-κΒ, οδηγεί στην αυξημένη έκφραση προ-φλεγμονωδών [παράγοντας νέκρωσης όγκου-α (TNFα), ιντερλευκίνη-6 (IL-6)] και προ-θρομβωτικών παραγόντων [ιστικού παράγοντα (TF)], συμβάλλοντας κατ’αυτό τον τρόπο στην εμφάνιση ενδοθηλιακής δυσλειτουργίας. Φραγμό στις δράσεις αυτές αποτελεί η διαλυτή μορφή του RAGE, sRAGE, η οποία συνδεόμενη με τα AGEs τα οδηγεί προς αποδόμηση και εμποδίζει την ενδοκυττάρια σηματοδότηση που πυροδοτούν τα AGEs. Σκοπός / μεθοδολογία: Προκειμένου να διερευνήσουμε τον πιθανό ρόλο των διεργασιών της φλεγμονής στην ανάπτυξη των διαβητικών επιπλοκών, σε μονοπύρηνα κύτταρα περιφερικού αίματος 47 προεφηβικών και εφηβικών παιδιών (ηλικίας 4-18 ετών) και 10 νεαρών ενηλίκων (ηλικίας 18-29 ετών) με ΣΔ1 και 39 μη διαβητικών μαρτύρων αντίστοιχης ηλικίας, μελετήσαμε: 1) με RT-PCR τη γονιδιακή έκφραση: του MCP-1, του CD68, της Akt2, του TNF-α, της IL-6 και του RAGE, 2) με ανοσοαποτύπωση κατά Western μετρήσαμε την πρωτεϊνική έκφραση της Akt2, του RAGE και του TF, 3) τα επίπεδα πλάσματος του sRAGE με τη μέθοδο της ELISA. Αποτελέσματα: Η μελέτη έδειξε ότι: (1) Το sRAGE: α) ήταν αυξημένο στους προεφηβικούς μάρτυρες σε σύγκριση με τους εφηβικούς και νεαρούς ενήλικες μάρτυρες, ενώ τα επίπεδα του ήταν μειωμένα στους προεφηβικούς διαβητικούς σε σύγκριση με τους αντίστοιχους μάρτυρες και β) είχε μία τάση μείωσης με τα αυξανόμενα επίπεδα της HbA1c. (2) Η γονιδιακή έκφραση του RAGE: α) έτεινε να αυξηθεί στους προεφηβικούς διαβητικούς και αυξανόταν σημαντικά στους νεαρούς ενήλικες διαβητικούς σε σύγκριση με τους αντίστοιχους μάρτυρες και β) σχετιζόταν αρνητικά με την HbA1c στους διαβητικούς ασθενείς >18ετών. (3) Η πρωτεϊνική έκφραση και των δύο ισομορφών του RAGE (46kDa και 80kDa) εμφάνισε μία τάση μείωσης με την εξέλιξη της εφηβείας (προεφηβικοί > εφηβικοί > νεαροί ενήλικες) στους μάρτυρες, μεταβολές που ακολούθησαν και οι διαβητικοί ασθενείς, αν και τα επίπεδα έκφρασης έτειναν να είναι πιο υψηλά. (4) Η 46kDa ισομορφή RAGE: α) ήταν αυξημένη στου νεαρούς ενήλικες διαβητικούς και μειωμένη στους διαβητικούς >13ετών με διάρκεια διαβήτη ≤5 έτη, σε σύγκριση με τους αντίστοιχους μάρτυρες αλλά και τους αντίστοιχης ηλικίας διαβητικούς με διάρκεια νόσου >5 έτη, β) έτεινε να αυξηθεί με τις τάξεις HbA1c: ≤7%-8% και να μειωθεί με τις τάξεις HbA1c: 8-10% και γ) συσχετίσθηκε θετικά με τη διάρκεια διαβήτη και με τα επίπεδα των τριγλυκεριδιών στο σύνολο των ασθενών. (5) Η 80kDa ισομορφή του RAGE: α) είχε μία τάση αύξησης στους διαβητικούς ασθενείς >13ετών με διάρκεια διαβήτη ≤5 έτη, σε σύγκριση με τους αντίστοιχους μάρτυρες και τους διαβητικούς αντίστοιχης ηλικίας αλλά διάρκεια νόσου >5έτη, β) συσχετίσθηκε αρνητικά με την HbA1c σε ασθενείς με διάρκεια νόσου ≤5έτη. (6) Το MCP-1: α) έτεινε να αυξηθεί στους διαβητικούς σε σύγκριση με τους μάρτυρες και συγκεκριμένα, η αύξηση ήταν σημαντική στους διαβητικούς με στάδιο εφηβείας ΙΙ και V, β) έτεινε να αυξηθεί με την HbA1c και αυξανόταν σημαντικά στους διαβητικούς με περίμετρο κοιλίας >75%. (7) Το CD68: α) βρέθηκε αυξημένο στα διαβητικά αγόρια >18 ετών σε σύγκριση με τους αντίστοιχους μάρτυρες, β) έτεινε να μειωθεί στους διαβητικούς >13ετών με διάρκεια διαβήτη >5έτη, σε σύγκριση με τους αντίστοιχης ηλικίας μάρτυρες και γ) έτεινε να μειωθεί με τα αυξανόμενα επίπεδα της HbA1c και στους διαβητικούς με LDL >100 mg/dL. (8) To TNF-α έδειξε: α) σημαντική μείωση στους διαβητικούς ασθενείς σε σχέση με τους μάρτυρες και ιδιαίτερα σε αυτούς που ήταν ≤13ετών με διάρκεια διαβήτη >5 έτη και σε αυτούς με ηλικία >18ετών, β) μία τάση αύξησης με τις τάξεις HbA1c μέχρι του 10%. (9) Η IL-6 έδειξε: α) σημαντική μείωση στους μάρτυρες με την παράλληλη αύξηση της ηλικίας, β) σημαντική αύξηση στους διαβητικούς >18ετών σε σύγκριση με τους αντίστοιχους μάρτυρες, γ) μία τάση μείωσης στους διαβητικούς ≤13ετών με διάρκεια διαβήτη >5έτη, σε σχέση με τους ηλικιακά αντίστοιχους μάρτυρες και δ) μία σημαντική μείωση στους διαβητικούς με HbA1c >10,1% σε σύγκριση με τις υπόλοιπες τάξεις της HbA1c. (10) Ο TF βρέθηκε: α) σημαντικά αυξημένος στα διαβητικά αγόρια ≤13 ετών και έτεινε να αυξηθεί στα διαβητικά αγόρια >18ετών σε σχέση με τους αντίστοιχους μάρτυρες, β) σημαντικά μειωμένος στους διαβητικούς με διάρκεια νόσου >5έτη και έτεινε να μειωθεί στους διαβητικούς με HbA1c >10,1% και σε αυτούς με LDL >100 mg/dL. (11) Η γονιδιακή έκφραση της Akt2 έδειξε: α) σημαντική αύξηση στους προεφηβικούς διαβητικούς, β) σημαντική μείωση στους διαβητικούς με LDL >100mg/dl και σε αυτούς με HbA1c >9%, σε σύγκριση με τους αντίστοιχους μάρτυρες. (12) Η πρωτεϊνική έκφραση της Akt2 έδειξε: α) μία τάση αύξησης στους προεφηβικούς διαβητικούς και στους διαβητικούς που βρίσκονται στις τάξεις HbA1c: 7,1%-9%, ενώ αυξανόταν σημαντικά στους διαβητικούς με διάρκεια διαβήτη >5έτη, β) σημαντική αύξηση στους διαβητικούς με LDL >100mg/dl.l Συμπέρασμα: Στα άτομα με ΣΔ1, φαίνεται να υπάρχει μια ήπια διαδικασία χημειοταξίας και ενεργοποίησης των μονοκυττάρων, ωστόσο η έκφραση των κυτταροκινών και των υπολοίπων μορίων που εμπλέκονται σε αυτή δεν είναι εντυπωσιακά αυξημένη σε σχέση με τους μάρτυρες. Το γεγονός αυτό μπορεί να σχετίζεται αφενός με τον καλό γλυκαιμικό και λιπιδαιμικό έλεγχο των ασθενών και αφετέρου στην τροποποιημένη ανοσολογική απόκριση που εμφανίζουν τα άτομα αυτά. / Diabetes Mellitus type 1 (DM1) is one of the most common types of diabetes in children and its prevalence has increased over the past decades. DM1 is usually associated with severe micro- and macro- angiopathic complications. The activation of monocytes, their chemotaxis (major chemotactic factor of monocytes: MCP-1) and their transformation to macrophages (marker of macrophages: CD68) is known to play a central role in the development of those complications. The redundant glucose, that characterizes DM1, reacts non enzymatically with plural substrates and causes the formation of AGEs (Advanced Glycation Endproducts). AGEs can cause tissue damage either by direct interaction with extracellular matrix’s domains or via their receptor (main representative: RAGE). RAGE’s activation, through the activation of the intracellular signaling pathways of protein kinase B (Akt2/PKB) and nuclear factor κB (NF-κB), leads to the increased expression of pro-inflammatory [Tissue Necrosis Factor-α (TNF-α), Interleukin-6 (IL-6)] and pro-thrombotic factors [Tissue Factor (TF)], contributing to the development of endothelial dysfunction. As a decoy to these actions of AGEs, the soluble form of RAGE, sRAGE, binds AGEs, leading them to degradation and therefore inhibiting the AGE-induced intracellular signaling. Aim / Material- Methods: In order to investigate the role of the inflammatory procedures in the development of diabetic complications, in peripheral blood mononuclear cells (PBMCs) of 47 pre-pubertal and pubertal children (4-18 years of age) and 10 young adults (18-29 years of age) with DM1 and 39 age-matched controls, we studied: 1) with RT-PCR the gene expression of: MCP-1, CD68, Akt2, IL-6 and RAGE, 2) with Western Immunoblotting the protein expression of Akt2, RAGE and TF, 3) sRAGE plasma levels were determined by ELISA. Results: Our study showed that: (1) sRAGE: a) was increased in pre-pubertal controls in comparison to the pubertal and the young adult controls, while its levels were decreased in pre-pubertal DM1 patients in comparison to their controls and b) tended to decrease with the progressive increase of HbA1c levels in DM1 patients. (2) RAGE gene expression: a) tended to increase in pre-pubertal DM1 patients and was significantly increased in young adults DM1 patients in comparison to their age-matched controls and b) was negatively correlated with HbA1c levels in DM1 patients >18years of age. (3) Protein expression of both RAGE isoforms (46kDa & 80kDa) tended to decrease with the progression of puberty (pre-pubertal > pubertal > young adults) in the control group, changes that were also found in DM1 patients, although the expression was at higher levels. (4) Protein expression of 46 kDa RAGE isoform: a) was increased in DM1 young adults and decreased in DM1 patients >13 years of age and duration of diabetes ≤5years, in comparison to age-matched controls and age-matched DM1 patients with duration of diabetes >5years, b) tended to increase with levels of HbA1c ≤7%-8% and decrease with levels of HbA1c 8-10% and c) was positively correlated with the duration of diabetes and the levels triglycerides in DM1 patients’ group. (5) Protein expression of 80kDa RAGE isoform: a) tended to increase in DM1 patients aged >13 years and with duration of diabetes ≤5 years, in comparison to age-matched controls and age-matched DM1 patients with duration of diabetes >5years, b) negatively correlated with HbA1c levels in DM1 patients with ≤5 years duration of diabetes. (6) MCP-1: tended to increase in DM1 patients in comparison to controls and this increase was significant in DM1 patients with II and V Tanner stage of puberty, b) tended to elevate with the progressive increase of HbA1c and was significantly higher in DM1 patients with waist circumference >75%. (7) CD68: a) was increased in DM1 boys >18 years of age in comparison to the age-matched controls, b) tended to decrease in DM1patients >13 years of age and duration of diabetes >5years, in comparison to the age-matched controls and c) tended to decrease with the increasing levels of HbA1c and also in DM1 patients with LDL >100 mg/dL. (8) TNF-α: a) was significantly decreased in DM1 patients in comparison to controls and especially to those with age ≤13 years and duration of diabetes >5years and them with >18 years of age, b) tended to increase with the increasing levels of HbA1c until 10%. (9) IL-6 showed: a) significant decrease in controls with the parallel increase of the age, b) significant increase in DM1 patients >18 years of age in comparison to the age-matched controls, c) a tendency to decrease in DM1 patients ≤13 years of age and duration of diabetes >5years, in comparison to the age-matched controls and d) significant decrease in DM1 patients with HbA1c levels >10,1%, in comparison to the rest of the determined levels of HbA1c. (10) TF: a) was significant elevated in DM1 boys ≤13 years of age and tended to increase in DM1 boys >18 years of age in comparison to the age-matched controls, b) was significant decreased in DM1 patients with duration of diabetes >5years and tended to decrease in DM1 patients with HbA1c levels >10,1% and them with LDL >100mg/dl. (11) Akt2 gene expression showed: a) significant increase in pre-bubertal DM1 patients, b) significant decrease in DM1 patients with LDL >100mg/dl and them with HbA1c levels >9%, in comparison to their controls. (12) Akt2 protein expression showed: a) a tendency to increase in pre-pubertal DM1 patients and in those with HbA1c levels between 7,1-9%, whereas it was increased significantly in DM1 patients with duration of diabetes >5years, b) a significant increase in DM1 patients with LDL >100mg/dl. Conclusions: In the DM1 patients studied, a low grade procedure of chemotaxis and activation of macrophages is possibly present, although the cytokines’ and inflammatory factors’ expression was not remarkably increased in comparison to the controls. This may reflect the good glycemic and lipidemic control of these patients who at the same time possibly exhibit small modifications in their immunological response.

Dissertação apresentada ao Programa de Pós-Graduação em Ciências da Saúde da Universidade do Extremo Sul Catarinense - UNESC, como requisito parcial para a obtenção do título de Mestre em Ciências da Saúde. / A meningite bacteriana é uma inflamação das meninges, podendo ser ocasionada pela bactéria Streptococcus pneumoniae que provoca uma intensa resposta imunológica no hospedeiro. Apesar dos avanços, esta doença ainda representa uma grande taxa de mortalidade e de morbidade no mundo inteiro. Os processos inflamatórios ativados durante a meningite pneumocócica aumentam a produção de citocinas e quimiocinas provocando uma cascata de mediadores inflamatórios, bem como a produção de espécies reativas de oxigênio (ERO), culminando na produção aumentada (e deposição) de proteínas amiloides. O aumento da produção e deposição do peptídeo β-amiloide, proporciona locais de ligação adicionais para receptor de produtos finais da glicação avançada (RAGE) nos neurônios e microglia. A interação do receptor RAGE com seu ligante (β-amiloide, por exemplo) é capaz de estimular múltiplas vias de sinalização intracelular, incluindo a ativação e a translocação para o núcleo do fator de transcrição NF-κB, que leva à produção de citocinas, quimiocinas, moléculas pró-inflamatórias, moléculas de adesão e indiretamente o estresse oxidativo. Isso pode levar a lesões neuronais causando prejuízo de memória e aprendizagem, bem como prejuízo a integridade da Barreira Hematoencefálica (BHE). Estratégias terapêuticas sugerem que RAGE é um importante alvo para o tratamento de doenças inflamatórias, visto que a inflamação aumenta a síntese de β-amiloide e regula positivamente RAGE, e a ativação dessa via induz disfunção cognitiva em longo prazo por apoptose neuronal. Recentes estudos sugerem que a inflamação no Sistema Nervoso Central (SNC) pode desempenhar um papel fundamental para o desenvolvimento de doenças neurodegenerativas. Portanto o objetivo deste trabalho foi determinar se a ativação da via β-amiloide-RAGE está associada à disfunção cognitiva em longo prazo em sobreviventes de meningite e se a modulação da inflamação com um inibidor específico de RAGE, o FPS-ZM1 iria melhorar esses efeitos. Em modelo experimental de meningite pneumocócica, foi avaliado a produção de citocinas: Fator de Necrose Tumoral α (TNF-α), interleucina 1β (IL-1β), e Interleucina 6 (IL6); RAGE e proteína do grupo 1 de alta mobilidade (HMGB1); integridade da BHE no hipocampo e no córtex pré-frontal; e também dano na memória de habituação e reconhecimento de objetos novos de ratos Wistar adultos 10 dias após o tratamento com FPS-ZM1. Nos testes a inibição do RAGE com o FPS-ZM1 preveniu a produção destas citocinas no córtex pré-frontal, diminuiu a quebra da BHE em 12, 18 e 24h tanto no hipocampo quanto no córtex, e diminuiu dano cognitivo 10 dias após a indução. Demonstrando um papel importante do RAGE na meningite causada pelo S. pneumoniae, tornando o FPS-ZM1 um possível tratamento adjuvante para esta doença, podendo prevenir danos neurológicos de longo prazo como apresentados na Doença de Alzheimer.

O RAGE é um receptor transmembrana, imunoglobulina-like que existe em múltiplas isoformas e interage com um amplo repertório de ligantes extracelulares. O RAGE é expresso em níveis baixos na maioria das células, porém o aumento da presença de seus ligantes no domínio extracelular faz com que o RAGE inicie uma cascata de sinalização intracelular complexa, resultando em estresse oxidativo, ativação do fator de transcrição NF-B, aumento da expressão de citocinas, além da indução de sua própria expressão. O envolvimento do RAGE neste amplo espectro de sinalização vincula este receptor a diversas condições patológicas. Nesta tese utilizamos 3 modelos experimentais que induzem inflamação sistêmica (Leishmania amazonensis, Lipopolissacarídeo e sepse) e 1 modelo experimental que mimetiza a denervação neuronal (modelos experimentais in vivo). Além disso utilizamos diferentes abordagens de bloqueio do RAGE a fim de elucidar a função deste receptor. Com base em nossos resultados os modelos experimentais foram eficientes em induzir o aumento do RAGE e sua sinalização no sistema nervoso central, desencadeando a síntese e liberação de moléculas pró-inflamatórias e o aumento do estresse oxidativo, culminando em neuroinflamação e neurodegeneração. As intervenções de bloqueio do RAGE foram eficientes em inibir as vias de sinalização intracelular mediadas pelo receptor, comprovando a via de ação. Levando em conta nossos principais resultados concluímos que: a) RAGE atua como mediador da perda neuronal em resposta ao insulto inflamatório em diversas estruturas do SNC, b) está presente no corpo dos neurônios dopaminérgicos e envolvido na morte destes neurônios; c) o aumento do RAGE é tempo-dependente e a morte dos neurônios está vinculada a ação deste receptor. / RAGE is a transmembrane, immunoglobulin-like receptor that exists in multiple isoforms and interacts with a broad repertoire of extracellular ligands. RAGE is expressed at low levels in most cells, but the increased presence of its ligands initiates a complex intracellular signaling cascade resulting in oxidative stress, activation of transcription factor NF-B, increased expression of cytokines in addition to concomitant upregulation of RAGE itself. In this thesis we used 3 experimental models which induce systemic inflammation (Leishmania amazonensis, Lipopolysaccharide and sepsis) and 1 experimental model that mimics neuronal denervation (experimental models in vivo). In addition, different approaches to block RAGE were used to elucidate the function of this receptor. Based on our results the experimental models were efficient in inducing the increase of RAGE and its signaling in the central nervous system, triggering the synthesis and release of proinflammatory molecules and the increase of oxidative stress, culminating in neuroinflammation and neurodegeneration. The RAGE blocking interventions were effective in inhibiting receptor-mediated signaling, proving the signaling pathway. Considering our main results, we conclude that: a) RAGE acts as a mediator of neuronal loss triggered by inflammatory insults in various CNS structures; b) RAGE is present in the body of dopaminergic neurons and is involved in the death of these neurons; c) the increase of RAGE is time-dependent, and the neuronal death is dependent on the action of this receptor.

O RAGE é um receptor transmembrana, imunoglobulina-like que existe em múltiplas isoformas e interage com um amplo repertório de ligantes extracelulares. O RAGE é expresso em níveis baixos na maioria das células, porém o aumento da presença de seus ligantes no domínio extracelular faz com que o RAGE inicie uma cascata de sinalização intracelular complexa, resultando em estresse oxidativo, ativação do fator de transcrição NF-B, aumento da expressão de citocinas, além da indução de sua própria expressão. O envolvimento do RAGE neste amplo espectro de sinalização vincula este receptor a diversas condições patológicas. Nesta tese utilizamos 3 modelos experimentais que induzem inflamação sistêmica (Leishmania amazonensis, Lipopolissacarídeo e sepse) e 1 modelo experimental que mimetiza a denervação neuronal (modelos experimentais in vivo). Além disso utilizamos diferentes abordagens de bloqueio do RAGE a fim de elucidar a função deste receptor. Com base em nossos resultados os modelos experimentais foram eficientes em induzir o aumento do RAGE e sua sinalização no sistema nervoso central, desencadeando a síntese e liberação de moléculas pró-inflamatórias e o aumento do estresse oxidativo, culminando em neuroinflamação e neurodegeneração. As intervenções de bloqueio do RAGE foram eficientes em inibir as vias de sinalização intracelular mediadas pelo receptor, comprovando a via de ação. Levando em conta nossos principais resultados concluímos que: a) RAGE atua como mediador da perda neuronal em resposta ao insulto inflamatório em diversas estruturas do SNC, b) está presente no corpo dos neurônios dopaminérgicos e envolvido na morte destes neurônios; c) o aumento do RAGE é tempo-dependente e a morte dos neurônios está vinculada a ação deste receptor. / RAGE is a transmembrane, immunoglobulin-like receptor that exists in multiple isoforms and interacts with a broad repertoire of extracellular ligands. RAGE is expressed at low levels in most cells, but the increased presence of its ligands initiates a complex intracellular signaling cascade resulting in oxidative stress, activation of transcription factor NF-B, increased expression of cytokines in addition to concomitant upregulation of RAGE itself. In this thesis we used 3 experimental models which induce systemic inflammation (Leishmania amazonensis, Lipopolysaccharide and sepsis) and 1 experimental model that mimics neuronal denervation (experimental models in vivo). In addition, different approaches to block RAGE were used to elucidate the function of this receptor. Based on our results the experimental models were efficient in inducing the increase of RAGE and its signaling in the central nervous system, triggering the synthesis and release of proinflammatory molecules and the increase of oxidative stress, culminating in neuroinflammation and neurodegeneration. The RAGE blocking interventions were effective in inhibiting receptor-mediated signaling, proving the signaling pathway. Considering our main results, we conclude that: a) RAGE acts as a mediator of neuronal loss triggered by inflammatory insults in various CNS structures; b) RAGE is present in the body of dopaminergic neurons and is involved in the death of these neurons; c) the increase of RAGE is time-dependent, and the neuronal death is dependent on the action of this receptor.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

A presente pesquisa teve como objetivo geral considerar os processos sociais envolvidos na situação do adolescente em conflito com a lei no Brasil, suas implicações na transformação de histórias de vidas e consequente demandas nos programas de atendimento. De forma mais específica, objetivou trazer conhecimentos concretos para o campo de trabalho socioeducativo, e investigar aquilo que se apresenta como propiciador da reincidência. Como forma de alcançar esses objetivos, foi considerada a seguinte questão norteadora: o que leva o adolescente a infracionar novamente? A metodologia foi baseada na Teoria do Vínculo de Pichon-Riviere, propondo-se como procedimento grupos operativos, os quais foram realizados com adolescentes do meio fechado (Internação) e meio aberto (Liberdade Assistida). O principal instrumento de coleta de dados consistiu no conteúdo das entrevistas, sendo a análise complementada com outras pesquisas realizadas na temática. Os dados coletados, considerando as medidas socioeducativas, demonstram que elas não dão conta do objeto ao qual se propõem: o rompimento com o ciclo da delinquência, com a repetição do ato de delinquir. A forma como as medidas estão sendo propostas são reguladoras muito externas, e não estão entrando no cerne do problema. Nesse sentido, devem considerar os sentimentos de raiva e ódio que movem o adolescente. No subtexto dessa raiva existem situações de humilhação e sentimentos de vergonha que, por não serem elaborados, criam o ciclo humilhação-vergonha-ódio. Entra-se em círculo odioso, retroalimentado, que supõe a raiva, a dimensão que o prazer de delinquir tem, ao que os adolescentes denominam adrenalina. Algo que os deixa aficionados como se fosse vício / The current research had as general purpose consider the social processes involved at the condition of adolescents in conflict with the law in Brazil, their implications at the process of changing of life histories and resultant demand in the service programs. More specifically, it intended bring real knowledge to the social educational field of work, and inquire into that present itself as reincidence propitiating. To achieve that objectives, it was considered the follow leading question: what does the adolescent commit an infraction again? The methology was based on the Theory of Bond by PICHON_RIVIERE, proposes as procedure operative groups, that was achieved with adolescents of closed system (internment) and opened system (Assisted Freedom). The main instrument to data collection was consisted in the interviews\'s content, being the analysis complemented with other researches achieved in the same area. The collected data, considering the social educational measures, demonstrate they can\'t handle of the object of their purpose: the break-up of deliquence cycle, with the recurrense of deliquence act. The way how these measures are being purpose are very external regulatories, and they are not moving into the heart of the matter. In this sense, they have to consider rage and hate feelings, that moves the adolescent. In the subtext of that hate there are abasement situations and feelings of shame that, because they are not elaborate, create a cycle of humiliation - shame - hate. Entering a hateful and feedback cycle, that suppose rage, the dimension of the pleasure of commit a crime, what they call of adrenaline. Which makes them excited, like an addiction

After the Glorious Revolution of 1688 the divisions within English society found additional expression through political parties as contemporaries staked out ideological positions on numerous issues and crises facing the nation. While the parties fought over issues of sovereignty and governance, the development of a taxation regime, required to pursue and support the nation’s almost constant wars on the continent, was also drawn into this contest. The nature of the debates over taxation on landed property provides an important lens through which to understand the ideological positions of both Whigs and Tories over matters of not only political economy, but religion, society, and governance. The English Land Tax, is one of the most important fiscal instruments of Augustan England and reveals how Whigs and Tories articulated positions on the aftermath of 1689, on the financial revolution that followed, and on the nature of governance at the beginning of the ‘long eighteenth century.’

La mortalité du diabète de type 2 est liée à ses complications cardiovasculaires (CVD). L'identification de nouveaux biomarqueurs associés à la dysfonction endothéliale, permettrait d'en améliorer le dépistage, la prévention. Au cours du diabète de type 2, l'hyperglycémie est associée à un fort stress oxydant. Nous nous sommes proposés ici d'évaluer l'impact de la glycoxydation sur la principale protéine circulante, l'albumine, et d'identifier si les modifications glycoxydatives de l'albumine dans le diabète avait un rôle dans la phyiopathologie des CVD du diabète de type 2. Nous avons pu mettre en évidence des modifications structurales et fonctionnelles importantes de l'albumine au cours du diabète de type 2 avec la formation entre autres de produits avancés de glycation (AGEs). Ces modifications glycoxydatives sont associées à des effets cellulaires pro-oxydant et pro-inflammatoire via une augmentation de l'expression du Récepteur aux AGEs (RAGE). Ces observations suggèrent que les formes glycoxydées d'albumine présentent un rôle central dans les mécanismes menant à la dysfonction endothélilale. Il reste néanmoins à évaluer l'intérêt du dosage des formes modifiées de l'albumine dans le dépistage des CVD au cours d'une étude clinique prospective de grande ampleur. Le développement d'une méthode de dosage rapide et reproductible des fractions d'albumine modifiées, comme celui de l'IMA (albumine modifiée par l'ischémie), en faciliterait la mise en œuvre. / Type 2 diabetes is dramatically associated with an enhanced cardiovascular complication risk. The identification of novel biomarkers associated with endothelial dysfunction remains highly warranted to improve diabetes screening and prevention. Oxidative stress and protein modifications are frequently observed in numerous disease states. Albumin, the major circulating protein in blood, can undergo increased glycoxidation in diabetes. Objectives of my thesis were to clarify the impact of glycoxidative modification of albumin on its structure and its functions and to determine whether such impairments may be encountered in albumin purified from diabetics. The occurrence of oxidative modifications was found to be enhanced in in vitro glycoxidized HAS and albumin purified from diabetics, after determination of their free thiol group content, relative electrophoretic migration, carbonyl content, and antioxidant activities. In addition, glycoxidized albumin exhibited impaired pharmaceutic molecule binding capacities. Cells treated with glycoxidized albumin exhibited a proinflammatory state attested by an overgeneration of intracellular reactive oxygen species, enhancements in RAGE expression, and an accumulation of carbonylated proteins.Methods to detect IMA (ischemia modified albumin) were developed and applied to diabetics patients. Relationships have been established between specific pathological parameters (cardiovascular disorders, hyperglycemia…) with an enhanced glycoxidative modification of albumin in diabetics. We thus propose that impaired albumin structure and function in relation in the enhanced oxidant stress observed in diabetics might be involved in the increased mortality risk of these patients.

This project will address the increasing problem of violence on our nations roadways. This form of violence, termed road rage, is also occurring worldwide.

Preeclampsia (PE), intrauterine growth restriction (IUGR) and gestational diabetes (GDM) increase the risk of maternal and fetal morbidity and mortality. The roles of Advanced Glycation End-products (AGEs) are already well documented concerning inflammation, hypoxia and oxidative stress. AGEs bind to its receptor, Receptor for Advanced Glycation End-products (RAGE), and activate an inflammatory pathway. This pathway alters the efficacy of invasive trophoblast cells and in the placenta and can result in placental dysfunction. We hypothesized that the placental dysfunction found in PE, IUGR, and GDM resulted from an over activation of the RAGE-mediated inflammatory pathway. Using human placental samples, we found that RAGE protein expression via western blotting was increased in PE and decreased in IUGR while GDM remained similar to that of control placentas. We then wanted to determine the efficacy of RAGE activation to alter the invasive nature of invasive cytotrophoblasts cells. We found that the addition of AGEs to SW71 cells decreases invasion through the activation of JNK and ERK cellular signaling pathways. Altogether these findings suggest that RAGE activation in trophoblast cells seems result in insufficient placental pathogenesis causing PE, however the IUGR and GDM samples we obtained did not seem to have resulted from RAGE activation. We also found that RAGE activation can alter the ability of invasive trophoblasts to invade, thus limiting the ability of the placental cells to remodel the maternal spiral arteries. We believe that further research into specific triggers of IUGR (smoking-induced) and un-treated diabetes could result in RAGE stimulated placental insufficiency.