Atherosclerosis is a macrophage-dominated nonresolving inflammatory disease of the arterial wall. Macrophage processes, including apoptosis, influence lesion development in atherosclerosis. Cannabinoids, compounds structurally related to Δ9-tetrahydrocannabinol (THC), the active ingredient in marijuana, exert their effects through cannabinoid receptors, CB1 and CB2. Cannabinoid treatment, THC or Win55,212-2, reduces atherosclerosis in ApoE-null mice by a mechanism thought to involve CB2. However, the exact role of CB2 in atherosclerosis remains unclear. We found that CB2-null macrophages are resistant to oxysterol/oxLDL-induced apoptosis leading us to hypothesize that CB2 may modulate macrophage apoptosis in atherosclerosis. To determine the functions of CB2 in atherosclerosis, we fed low density lipoprotein receptor-null (Ldlr-/-) and Ldlr-/- mice genetically deficient in CB2, an atherogenic diet for 8 and 12 weeks. CB2 deficiency did not significantly affect aortic root lesion area after 8 or 12 weeks; however, after 12 weeks, CB2-deficient lesions displayed increased lesional macrophage and smooth muscle cell (SMC) content and a ~2-fold reduction in lesional apoptosis. CB2-deficienct lesions also displayed reduced collagen content and elevated elastin fiber fragmentation that was associated with elevated levels of the extracellular matrix degrading enzyme, matrix metalloproteinase 9 (MMP9). These results demonstrate that although CB2 signaling does not affect atherosclerotic lesion size it does modulate lesional apoptosis, cellularity and ECM composition. Ldlr-/- and CB2-deficient Ldlr-/- mice were also subjected to daily treatments with Win55,212-2, a synthetic cannabinoid, over the last 2 weeks of an 8 week atherogenic diet to identify CB2-dependent and CB2-independent effects of cannabinoid receptor stimulation on atherosclerosis. Win55,212-2 did not affect hypercholesterolemia, aortic root lesion area, lesional macrophage infiltration, or ECM composition in either genotype but did significantly reduce total plasma triglyceride levels and lesional SMC content, independent of CB2. Surprisingly, lesional apoptosis was dose-dependently repressed by Win55,212-2 in Ldlr-/- mice by a CB2-dependent mechanism. All together, these results support the suggestion that CB2 may be a target for novel therapies aimed at modulating lesional apoptosis and cellularity to increase lesion stability and reduce the vulnerability to rupture.

Le PCK3145 est un peptide de 15 acides aminés inhibant la sécrétion de MMP-9 et démontrant une activité anti-tumorale contre le cancer de la prostate. Comme les cancers hématologiques sécrètent MMP-9, nous avons donc évalué l’effet du PCK3145 sur ces cancers. Nous avons démontré que les lignées humaines de lymphome non- Hodgkinien (LNH) SR et de myélome multiple RPMI-8226 ainsi que la lignée murine de mastocytome P815 ont une prolifération réduite suite à une exposition au PCK3145. Ce peptide diminue également la clonogénicité de ces cellules. In vivo, le PCK3145 diminue significativement la croissance des tumeurs sous-cutanées P815 comparativement au PBS (p<0.001) et aux peptides contrôles (« scrambled peptide » (p<0.05) et PCK5266 (p<0.01)). De plus, le traitement au PCK3145 diminue le nombre de métastases au niveau du foie par rapports aux contrôles (p<0.05). Les niveaux de MMP-9 dans le sang des souris traitées au PCK3145 sont similaires à ceux dans le sang des souris sans tumeur. Par contre, chez les souris recevant le PBS ou le « scrambled peptide », les niveaux de MMP-9 étaient significativement plus élevés que dans les souris sans tumeur et les souris traitées au PCK3145 (p<0.05). De surcroît, dans un modèle de xénogreffe, le PCK3145 diminue significativement la croissance des lymphomes SR par rapport au PBS (p<0.01) et au « scrambled peptide » (p<0.001). Ces résultats indiquent que le PCK3145 possède une activité anti-tumorale et pourrait représenter un agent intéressant pour le traitement de plusieurs cancers hématologiques. / PCK3145 has been shown to exert anti-tumor activity against prostate cancer cells. In a Phase I clinical study, this peptide demonstrated low toxicity. To determine whether PCK3145 could exert cytotoxic activity against other marrow infiltrating cancers, we tested its activity against hematologic cancers. Interestingly, PCK3145 inhibited the proliferation of human NHL (SR) and myeloma (RPMI-8226) cell lines and murine mastocytoma (P815) cell line in vitro. Moreover, PCK3145 reduced the clonogenicity of these cell lines. To explore its activity in vivo, DBA/2 mice were injected with P815 cells. PCK3145 treatment significantly decreased P815 tumors growth in comparison to PBS (p<0.001), scrambled peptide (p<0.05) and PCK5266 (amino acids 52-66 of PSP-94) (p<0.01). Intraperitoneal PCK3145 treatment led to a decreased number of liver metastasis compared to PBS (p<0.05) and scrambled peptide (p<0.05). MMP-9 levels, measured by ELISA, in the peripheral blood of treated P815 bearing mice were similar to those obtained with healthy animals (12.83 1.890 (mean SD) ng/ml and 6.48 0.4070 ng/ml, respectively), while MMP-9 levels were elevated in mice treated with PBS and scrambled peptide (35.12 8.559 ng/ml and 22.60 3.944 ng/ml, respectively; p<0.05). In NOD/SCID mice PCK3145 treatment resulted in significant inhibition of human NHL SR growth compared to treatment with PBS (p<0.001) and scrambled peptide (p<0.01). Consequently, treatment with PCK3145 can reduce tumor cell proliferation of murine and human hematologic cancers. In addition, PCK3145 has the potential to inhibit tumor cells dissemination by lowering MMP-9 secretion. Thus, PCK3145 represents a unique peptide demonstrating sequence-specific anti-tumor activity hematologic malignancies.

Introdução: Aneurisma de aorta é considerado uma doença inflamatória crônica, mas ainda é controverso se está relacionado a aterosclerose aos agentes infecciosos. Antígenos de Chlamydophila pneumoniae (CP) e Mycoplasma pneumoniae (MP) foram encontrados em grande quantidade nas placas instáveis que usualmente estão associadas a remodelamento positivo e inflamação do vaso. Metaloproteinase da matriz 9 (MMP-9) está implicada na fragilidade da parede vascular e na formação dos aneurismas. Os efeitos imunomodulatórios dos receptores ativados por proliferador de peroxissomo (PPARs) têm sido relacionados à aterosclerose. Objetivos: Comparar lesões ateroscleróticas graves com e sem aneurisma do ponto de vista inflamatório e infeccioso, analisando antígenos de MP e CP e expressão de MMP-9, TIMP-1 e PPARs. Métodos: No presente estudo quantificamos, através da técnica de imunoistoquímica, antígenos de MMP- 9, TIMP-1, PPARs a e , e dos agentes infecciosos CP e MP em fragmentos de aorta ateroscleróticos com aneurisma (n=14) e sem aneurisma (n=14). Resultados: A adventícia e o tecido adiposo periadventicial (TAP) dos aneurismas apresentaram intensa inflamação. Expressão de MMP-9 esteve aumentada no TAP e agentes infecciosos, TIMP-1 e PPAR a estiveram aumentados na adventícia e no TAP, sem diferença em relação a expressão de PPAR . Colágeno, TIMP-1 e PPARs estiveram positivamente correlacionados no grupo com aterosclerose, mas não no grupo com aneurisma. Conclusão: Nossos achados sugerem que aneurisma de aorta é uma complicação das lesões ateroscleróticas. Quantidade aumentada de Chlamydophila pneumoniae e Mycoplasma pneumoniae na adventícia e PAT sem correlação da resposta de TIMP-1 e PPARs são achados que estão associados com a presença de aneurisma nos segmentos de aorta ateroscleróticos / Introduction: Aortic aneurysm is considered a chronic inflammatory disease, but remains a controversial matter if it is related to atherosclerosis and infectious agents. Chlamydophila pneumoniae (CP) and Mycoplasma pneumoniae (MP) antigens were found in higher amount in unstable plaques that are usually associated to positive remodeling and vessel inflammation. Matrix metalloproteinase 9 (MMP-9) has been implicated in vascular wall fragility and aneurysm development. The immunemodulator effects of peroxisome proliferators - activated receptor (PPARs) have been related to atherosclerosis. Objectives: To compare severe atherosclerotic lesions with and without aneurysms by inflammatory and infectious point of view, analyzing MP and CP, MMP-9, TIMP-1 and PPARs antigens. Methods: In the present study we quantified, using immunohistochemistry technique, MMP-9, TIMP-1, PPAR a and and infectious agents CP and MP antigens in aortic atherosclerotic fragments with aneurysms. (n=14) and without aneurysms (n=14). Results: Adventitia and periadventitial adipose tissue (PAT) from aneurysms showed intense inflammation. MMP-9 expression has increased in PAT and the infectious agents, TIMP-1 and PPAR a were increased in adventitia and PAT in aneurysmatic group, without difference related to PPAR expression. Collagen, TIMP-1 and PPARs were positively correlated in atherosclerotic group, but it was not observed in aneurysmatic group. Conclusion: Our data have suggested that aortic aneurysm is a complication of aortic atherosclerotic lesions. Increased amount of Chlamydophila pneumoniae and Mycoplasma pneumoniae in the adventitia and PAT without a correlated TIMP-1 and PPARs response are findings that were associated with the presence of aneurysm in atherosclerotic aortic segments

INTRODUÇÃO: Recidivas locais e loco-regionais são as principais causas de falha do tratamento em pacientes portadores de carcinomas epidermóides de boca e orofaringe. A cirurgia de resgate é geralmente a melhor opção terapêutica para esses pacientes. Esse estudo tem por objetivo avaliar a importância prognóstica da expressão das proteínas EGFR, MMP-2, MMP-9 e VEGF em pacientes com recidiva local submetidos à cirurgia de resgate. CASUÍSTICA E MÉTODOS: Os prontuários de 111 pacientes portadores de recorrência local de carcinomas epidermóides de boca e orofaringe foram analisados de forma retrospectiva. A localização do tumor primário foi o lábio em 10 casos (9%), a cavidade oral em 68 (61%) e a orofaringe em 33 (30%). O tratamento prévio foi cirurgia em 33 casos (30%), radioterapia associada ou não à quimioterapia baseada em cisplatina em 46 (41%) e cirurgia com radioterapia adjuvante em 32 (29%). A expressão das proteínas EGFR, MMP-2, MMP-9 and VEGF foi avaliada com a técnica do Tissue Microarray. RESULTADOS: O intervalo livre de doença variou de 0,89 a 140,9 meses, com uma mediana de 6,87 meses. As recidivas foram diagnosticadas em intervalo de tempo inferior a 1 ano em 69 pacientes (62,2%) e após 1 ano em 42 (37,8%). Os pacientes com intervalo livre de doença inferior a 1 ano apresentaram pior resultado de sobrevida (p=0,01). O estádio clínico da recidiva (rEC) foi I ou II em 31 casos (27,9%) e III ou IV em 80 (72,1%). Pacientes com doença em estádio clínico mais avançado (rEC III ou IV) apresentaram piores taxas de sobrevida específica por câncer (p=0,04). Hiper-expressão do EGFR foi associada a pior resultado do tratamento. Os casos com EGFR positivo obtiveram sobrevida específica por câncer em 3 anos de 27,2%, enquanto pacientes com EGFR negativo alcançaram 64,3% de sobrevida em 3 anos (p=0,001). A expressão das proteínas MMP-2, MMP-9 e VEGF não se mostrou significativa para o prognóstico (p=0,83, p=0,15 e p=0,86, respectivamente). Na análise multivariada, apenas o intervalo livre de doença e a expressão do EGFR foram associadas à maior risco de morte. CONCLUSÕES: Recidivas locais de carcinomas epidermóides de boca e orofaringe são associadas a mau prognóstico. Intervalo livre de doença superior a 1 ano e ausência de expressão do EGFR foram os principais fatores associados a melhores resultados de sobrevida específica por câncer em pacientes submetidos à cirurgia de resgate. / INTRODUCTION: Local and regional relapses are the main sites of treatment failure in patients with oral and oropharyngeal squamous cell carcinoma. In these instances, salvage surgery is the most widely used treatment approach. The aim of this study is to analyze the prognostic effect of the expression of EGFR, MMP-2, MMP-9 and VEGF in patients with recurrent cancer sumitted to salvage surgery. METHODS: The charts of 111 patients with local recurrence of oral or oropharyngeal squamous cell carcinomas were retrospectively analyzed. The tumor sites were: the lip in 11 cases (9%), the oral cavity in 68 (61%) and the oropharynx in 33 (30%). The previous treatment was: Surgery in 33 patients (30%), radiotherapy with or without cisplatin based chemotherapy in 46 (41%) and surgery with adjuvant radiotherapy in 32 (29%). EGFR, MMP-2, MMP-9 and VEGF expressions were analyzed with tissue microarray immunohistochemical technique. RESULTS: The disease-free interval ranged from 0.89 to 140.9 months with a median of 6.87 months. The patients were categorized into two groups: Those with recurrence in less than 1 year (69 patients - 62.2%) and those with recurrence after 1 year (42 - 37.8%). The group with the shorter disease-free interval presented a worse prognosis (p=0.01). The clinical stage of recurrence (rCS) was I/II in 31 cases (27.9%) and III/IV in 80 cases (72.1%). Patients with more advanced diseases (rCS III/IV) had worse rates of cancer specific survival (CSS) than patients with rCS I/II (p=0.04). An over-expression of EGFR was associated with worse treatment results. Positive EGFR cases had a 3 year CSS of 27.2%, while EGFR negative patients had 64.3% (p=0.001). The MMP-2 and MMP-9 over-expression were also associated with a worse prognosis but without statistical significance (p=0.83 and p=0.15). VEGF expression did not show prognostic significance in this group of patients. In a multivariate analysis only the disease-free interval and over-expression of EGFR were associated with a higher risk of death. CONCLUSION: Local recurrence in oral and oropharyngeal squamous cell carcinomas usually indicates an unfavorable prognosis. A disease-free interval greater than 1 year and a negative EGFR expression are the main prognostic factors which indicate a better cancer specific survival rate in patients submitted to salvage surgery.

INTRODUÇÃO: Recidivas locais e loco-regionais são as principais causas de falha do tratamento em pacientes portadores de carcinomas epidermóides de boca e orofaringe. A cirurgia de resgate é geralmente a melhor opção terapêutica para esses pacientes. Esse estudo tem por objetivo avaliar a importância prognóstica da expressão das proteínas EGFR, MMP-2, MMP-9 e VEGF em pacientes com recidiva local submetidos à cirurgia de resgate. CASUÍSTICA E MÉTODOS: Os prontuários de 111 pacientes portadores de recorrência local de carcinomas epidermóides de boca e orofaringe foram analisados de forma retrospectiva. A localização do tumor primário foi o lábio em 10 casos (9%), a cavidade oral em 68 (61%) e a orofaringe em 33 (30%). O tratamento prévio foi cirurgia em 33 casos (30%), radioterapia associada ou não à quimioterapia baseada em cisplatina em 46 (41%) e cirurgia com radioterapia adjuvante em 32 (29%). A expressão das proteínas EGFR, MMP-2, MMP-9 and VEGF foi avaliada com a técnica do Tissue Microarray. RESULTADOS: O intervalo livre de doença variou de 0,89 a 140,9 meses, com uma mediana de 6,87 meses. As recidivas foram diagnosticadas em intervalo de tempo inferior a 1 ano em 69 pacientes (62,2%) e após 1 ano em 42 (37,8%). Os pacientes com intervalo livre de doença inferior a 1 ano apresentaram pior resultado de sobrevida (p=0,01). O estádio clínico da recidiva (rEC) foi I ou II em 31 casos (27,9%) e III ou IV em 80 (72,1%). Pacientes com doença em estádio clínico mais avançado (rEC III ou IV) apresentaram piores taxas de sobrevida específica por câncer (p=0,04). Hiper-expressão do EGFR foi associada a pior resultado do tratamento. Os casos com EGFR positivo obtiveram sobrevida específica por câncer em 3 anos de 27,2%, enquanto pacientes com EGFR negativo alcançaram 64,3% de sobrevida em 3 anos (p=0,001). A expressão das proteínas MMP-2, MMP-9 e VEGF não se mostrou significativa para o prognóstico (p=0,83, p=0,15 e p=0,86, respectivamente). Na análise multivariada, apenas o intervalo livre de doença e a expressão do EGFR foram associadas à maior risco de morte. CONCLUSÕES: Recidivas locais de carcinomas epidermóides de boca e orofaringe são associadas a mau prognóstico. Intervalo livre de doença superior a 1 ano e ausência de expressão do EGFR foram os principais fatores associados a melhores resultados de sobrevida específica por câncer em pacientes submetidos à cirurgia de resgate. / INTRODUCTION: Local and regional relapses are the main sites of treatment failure in patients with oral and oropharyngeal squamous cell carcinoma. In these instances, salvage surgery is the most widely used treatment approach. The aim of this study is to analyze the prognostic effect of the expression of EGFR, MMP-2, MMP-9 and VEGF in patients with recurrent cancer sumitted to salvage surgery. METHODS: The charts of 111 patients with local recurrence of oral or oropharyngeal squamous cell carcinomas were retrospectively analyzed. The tumor sites were: the lip in 11 cases (9%), the oral cavity in 68 (61%) and the oropharynx in 33 (30%). The previous treatment was: Surgery in 33 patients (30%), radiotherapy with or without cisplatin based chemotherapy in 46 (41%) and surgery with adjuvant radiotherapy in 32 (29%). EGFR, MMP-2, MMP-9 and VEGF expressions were analyzed with tissue microarray immunohistochemical technique. RESULTS: The disease-free interval ranged from 0.89 to 140.9 months with a median of 6.87 months. The patients were categorized into two groups: Those with recurrence in less than 1 year (69 patients - 62.2%) and those with recurrence after 1 year (42 - 37.8%). The group with the shorter disease-free interval presented a worse prognosis (p=0.01). The clinical stage of recurrence (rCS) was I/II in 31 cases (27.9%) and III/IV in 80 cases (72.1%). Patients with more advanced diseases (rCS III/IV) had worse rates of cancer specific survival (CSS) than patients with rCS I/II (p=0.04). An over-expression of EGFR was associated with worse treatment results. Positive EGFR cases had a 3 year CSS of 27.2%, while EGFR negative patients had 64.3% (p=0.001). The MMP-2 and MMP-9 over-expression were also associated with a worse prognosis but without statistical significance (p=0.83 and p=0.15). VEGF expression did not show prognostic significance in this group of patients. In a multivariate analysis only the disease-free interval and over-expression of EGFR were associated with a higher risk of death. CONCLUSION: Local recurrence in oral and oropharyngeal squamous cell carcinomas usually indicates an unfavorable prognosis. A disease-free interval greater than 1 year and a negative EGFR expression are the main prognostic factors which indicate a better cancer specific survival rate in patients submitted to salvage surgery.

Os tumores hipofisários mesmo sendo, em sua maioria, benignos, podem apresentar comportamento invasivo, com extensão para seio cavernoso, seio esfenoidal e clivo. As metaloproteinases de matriz tipo 2 (MMP-2) e 9 (MMP-9) e o inibidor tecidual de metaloproteinases-2 (TIMP-2) têm sido estudados em relação ao comportamento invasivo desses tumores, em especial quanto à invasão do seio cavernoso. Esse estudo teve como objetivo avaliar a expressão proteica das MMP-2, MMP-9 e do TIMP-2 nos tumores hipofisários e sua relação com invasão do seio cavernoso e, de maneira inédita, investigar a expressão dessas proteínas em nível plasmático. Adicionalmente, foram avaliadas a expressão dos RNAs mensageiros (RNAm) das MMP-2 e TIMP-2 com intuito de correlacioná-las com a expressão proteica no tecido tumoral, bem como a expressão do marcador de proliferação Ki67. Foram selecionados 77 casos, todos com amostras de tumor emblocado em parafina para análise imuno-histoquímica (IHQ). Destes, foram coletadas amostras de tumor fresco em 29 pacientes e de sangue periférico pré-operatório em outros 29 casos. A expressão proteica plasmática foi detectada de forma semi-quantitativa utilizando um arranjo de anticorpos em membrana comercial. A expressão dos RNAm das MMP-2 e TIMP-2 foi avaliada por reação em cadeia da polimerase quantitativo (qPCR) em tempo real. Do total de casos, 20 pacientes apresentavam tumores com invasão para o seio cavernoso. A expressão proteica tumoral das MMP-2, MMP-9 e TIMP-2 apresentou-se aumentada no grupo invasivo, contudo esta diferença não foi estatisticamente significante em relação ao grupo não- invasivo. A expressão plasmática das MMP-9 e TIMP-2 também não mostrou diferença entre os dois grupos e não se correlacionou com a expressão tumoral. A expressão plasmática da MMP-2 não foi detectada em nenhum caso. Quanto à expressão do RNAm das MMP-2 e TIMP-2, também não houve diferença significante entre os grupos e nem correlação com a expressão proteica tecidual ou plasmática. Foi observada uma diferença significante na dimensão tumoral [3.6 (2.5-5.2) x 2.0 (1.3-2.7); P < 0.001] e no índice do Ki67 [1.05 (0.27-25) x 0.5 (0.2-1.0); P < 0.001] entre os grupos invasivo e não-invasivo respectivamente. Em conclusão, em nossa coorte, não foi encontrada relação entre a expressão tecidual e plasmática das MMP-2, MMP-9 e TIMP-2 e a invasão para o seio cavernoso nos adenomas hipofisários / Pituitary tumors, although mostly benign, may present invasive behavior, with extension to the cavernous sinus, sphenoid sinus and clivus. Type 2 (MMP-2) and type 9 (MMP-9) matrix metalloproteinases and the metalloproteinase tissue inhibitor type 2 (TIMP-2) have been studied in relation to the invasive behavior of these tumors, especially regarding invasion of the cavernous sinus. The aim of this study was to evaluate the protein expression of MMP-2, MMP-9 and TIMP-2 in pituitary tumors and its relation with invasion of the cavernous sinus and, in an unprecedented way, to investigate the expression of these proteins at the plasma level. Additionally, expression of MMP-2 and TIMP-2 messenger RNAs (mRNAs) was evaluated in order to correlate with protein expression in tumor tissue, as well as Ki67 proliferation marker expression. A total of 77 cases were selected, all of them with paraffin embedded tumor samples for immunohistochemical analysis (IHC). Of these, fresh tumor samples were collected in 29 patients and preoperative peripheral blood in another 29 cases. Protein plasma expression was detected semi-quantitatively using a commercial membrane antibody array. Expression of MMP-2 and TIMP-2 mRNAs was evaluated by quantitative realtime polymerase chain reaction (qPCR). Of the total cases, 20 patients presented tumors invasive to the cavernous sinus. Tumor protein expression of MMP-2, MMP-9 and TIMP-2 was increased in the invasive group, not reaching, however, statistically significant difference as compared with the non-invasive group. Plasma expression of MMP-9 and TIMP-2 also did not differ between the two groups and did not correlate with tumor expression. Plasma expression of MMP-2 was not detected in any case. Concerning MMP-2 and TIMP-2 mRNA expression, there was also no significant difference between groups and no correlation with tissue or plasma protein expression was observed. A significant difference was observed in tumor size [3.6 (2.5-5.2) x 2.0 (1.3-2.7); P < 0.001] and in the Ki67 index [1.05 (0.27-25) x 0.5 (0.2-1.0); P < 0.001] between the invasive and non-invasive groups respectively. In conclusion, in our cohort, no relationship was found between the tissue and plasma expression of MMP-2, MMP-9 and TIMP-2 and the invasion of the cavernous sinus in pituitary adenomas

Scar-free wound healing and regeneration are uncommon phenomena permitting the near complete restoration of damaged tissues, organs and structures. Although rare in mammals, many lizards are able to undergo scarless healing and regeneration following loss of the tail. This study investigated the spontaneous and intrinsic capacity of the leopard gecko (Eublepharis macularius) tail to undergo scar-free wound healing and regeneration following two different forms of tail loss: autotomy, a voluntary and evolved mechanism of tail shedding at fracture planes; and surgical amputation, involuntary loss of the tail outside the fracture planes. Furthermore, I investigated the ability of the regenerate tail to regenerate by amputating a regenerate tail (previously lost by autotomy). To investigate these phenomena I imaged wound healing and regenereating tails daily (following autotomy and amputation) to document gross morphological changes. I used histochemistry to document tissue structure and immunohistochemistry to determine the tissue/cellular location of my five proteins of interest (PCNA, MMP-9, WE6, α-sma, TGF-β3). Each of these proteins of interest has been previously documented during wound healing and/or regeneration in other wound healing/regeneration model organisms (e.g. mice, urodeles, lizards, zebrafish). Scar-free wound healing and regeneration occurred following autotomy, amputation of the original tail and amputation of the regenerate tail, indicating that the leopard gecko tail has an instrinsic scar-free wound healing and regenerative capacity that is independent of the mode of tail loss (autotomy or amputation). Furthermore immunohistochemistry revealed a conserved sequence and location of the expression of the five proteins of interest following both forms of tail loss. These results provide the basis for further studies investigating scar-free wound healing and regeneration in a novel amniote model, the leopard gecko. / NSERC

Le PCK3145 est un peptide de 15 acides aminés inhibant la sécrétion de MMP-9 et démontrant une activité anti-tumorale contre le cancer de la prostate. Comme les cancers hématologiques sécrètent MMP-9, nous avons donc évalué l’effet du PCK3145 sur ces cancers. Nous avons démontré que les lignées humaines de lymphome non- Hodgkinien (LNH) SR et de myélome multiple RPMI-8226 ainsi que la lignée murine de mastocytome P815 ont une prolifération réduite suite à une exposition au PCK3145. Ce peptide diminue également la clonogénicité de ces cellules. In vivo, le PCK3145 diminue significativement la croissance des tumeurs sous-cutanées P815 comparativement au PBS (p<0.001) et aux peptides contrôles (« scrambled peptide » (p<0.05) et PCK5266 (p<0.01)). De plus, le traitement au PCK3145 diminue le nombre de métastases au niveau du foie par rapports aux contrôles (p<0.05). Les niveaux de MMP-9 dans le sang des souris traitées au PCK3145 sont similaires à ceux dans le sang des souris sans tumeur. Par contre, chez les souris recevant le PBS ou le « scrambled peptide », les niveaux de MMP-9 étaient significativement plus élevés que dans les souris sans tumeur et les souris traitées au PCK3145 (p<0.05). De surcroît, dans un modèle de xénogreffe, le PCK3145 diminue significativement la croissance des lymphomes SR par rapport au PBS (p<0.01) et au « scrambled peptide » (p<0.001). Ces résultats indiquent que le PCK3145 possède une activité anti-tumorale et pourrait représenter un agent intéressant pour le traitement de plusieurs cancers hématologiques. / PCK3145 has been shown to exert anti-tumor activity against prostate cancer cells. In a Phase I clinical study, this peptide demonstrated low toxicity. To determine whether PCK3145 could exert cytotoxic activity against other marrow infiltrating cancers, we tested its activity against hematologic cancers. Interestingly, PCK3145 inhibited the proliferation of human NHL (SR) and myeloma (RPMI-8226) cell lines and murine mastocytoma (P815) cell line in vitro. Moreover, PCK3145 reduced the clonogenicity of these cell lines. To explore its activity in vivo, DBA/2 mice were injected with P815 cells. PCK3145 treatment significantly decreased P815 tumors growth in comparison to PBS (p<0.001), scrambled peptide (p<0.05) and PCK5266 (amino acids 52-66 of PSP-94) (p<0.01). Intraperitoneal PCK3145 treatment led to a decreased number of liver metastasis compared to PBS (p<0.05) and scrambled peptide (p<0.05). MMP-9 levels, measured by ELISA, in the peripheral blood of treated P815 bearing mice were similar to those obtained with healthy animals (12.83 1.890 (mean SD) ng/ml and 6.48 0.4070 ng/ml, respectively), while MMP-9 levels were elevated in mice treated with PBS and scrambled peptide (35.12 8.559 ng/ml and 22.60 3.944 ng/ml, respectively; p<0.05). In NOD/SCID mice PCK3145 treatment resulted in significant inhibition of human NHL SR growth compared to treatment with PBS (p<0.001) and scrambled peptide (p<0.01). Consequently, treatment with PCK3145 can reduce tumor cell proliferation of murine and human hematologic cancers. In addition, PCK3145 has the potential to inhibit tumor cells dissemination by lowering MMP-9 secretion. Thus, PCK3145 represents a unique peptide demonstrating sequence-specific anti-tumor activity hematologic malignancies.

Made available in DSpace on 2015-07-22T20:49:30Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-24. Added 1 bitstream(s) on 2015-08-11T03:26:30Z : No. of bitstreams: 1 Publico-429a.pdf: 1596515 bytes, checksum: 2107c026687c3e2ecc62f0a5fffa1409 (MD5). Added 1 bitstream(s) on 2015-08-11T03:26:30Z : No. of bitstreams: 2 Publico-429a.pdf: 1596515 bytes, checksum: 2107c026687c3e2ecc62f0a5fffa1409 (MD5) Publico-429b.pdf: 1614171 bytes, checksum: d98715f7a666adc5ceb185cce978000e (MD5). Added 1 bitstream(s) on 2015-08-11T03:26:30Z : No. of bitstreams: 3 Publico-429a.pdf: 1596515 bytes, checksum: 2107c026687c3e2ecc62f0a5fffa1409 (MD5) Publico-429b.pdf: 1614171 bytes, checksum: d98715f7a666adc5ceb185cce978000e (MD5) Publico-429c.pdf: 1500700 bytes, checksum: 77791620b06c9404d28496c4f4ca3dc3 (MD5). Added 1 bitstream(s) on 2015-08-11T03:26:30Z : No. of bitstreams: 4 Publico-429a.pdf: 1596515 bytes, checksum: 2107c026687c3e2ecc62f0a5fffa1409 (MD5) Publico-429b.pdf: 1614171 bytes, checksum: d98715f7a666adc5ceb185cce978000e (MD5) Publico-429c.pdf: 1500700 bytes, checksum: 77791620b06c9404d28496c4f4ca3dc3 (MD5) Publico-429d.pdf: 1826591 bytes, checksum: 70c3dc706cb97372e3f26a7452c01208 (MD5) / O Linfoma de Hodgkin clássico (LHC) é caracterizado pela presença de uma pequena população de células grandes mono ou multinucleadas, denominadas células de Hodgkin-Reed-Sternberg (HRS), circundadas por uma grande massa inflamatória de células não neoplásicas. O vírus Epstein-Barr (EBV) está associado ao Linfoma de Hodgkin em cerca de 50% dos casos. O diagnóstico do LH EBV relacionado é possível por meio da identificação de proteínas virais nas células HRS. Os métodos considerados ideais para essa identificação são as reações de imuno-histoquímica utilizando anticorpos antiproteína latente de membrana (LMP1) e hibridação in situ com uma sonda para o RNA viral (EBER). A LMP-1 é considerada um oncogene clássico. Foi demonstrado que a LMP-1 pode controlar a expressão do gene da metaloproteinase 9 (MMP-9), em linhagem de células C33A. A MMP-9 é um membro da família das endopeptidases que facilita a invasão tumoral e metástases pela degradação do estroma extracelular. Objetivos: avaliar se a expressão da MMP-9 está relacionada ao status do EBV no tumor e se houve impacto na sobrevida livre de eventos (SLE) e sobrevida global (SG) em pacientes com LHC. Casuística e Métodos: foram examinados 97 pacientes com LHC. Todos os pacientes foram submetidos a protocolos de tratamentos equivalentes (MOPPABV ou ABVD). O diagnóstico histopatológico foi revisto e o subtipo classificado de acordo com a OMS. Reações de imuno-histoquímica para LMP-1 e MMP-9 e hibridação in situ para EBER foram realizadas. Resultados: A presença do EBV foi identificada em 52,5% dos casos. Houve uma maior prevalência do subtipo histológico celularidade mista em pacientes EBV positivos (P = 0,005). Não houve diferença na positividade do EBV em relação à faixa etária, sexo, estádio da doença ou pela presença de sintomas B. A presença do EBV no LHC não influenciou a SLE (P = 0,38) ou a SG (P = 0,80) com uma mediana de acompanhamento de 71 meses. A expressão da MMP-9 ocorreu em 87,6% dos casos estudados. Não houve diferença de casos positivos e negativos em relação ao status do EBV (P = 0,59). Quando avaliada a intensidade da expressão da MMP-9 nos casos positivos também não observamos correlação com a presença do EBV (P = 0,62). Não houve diferença entre o resultado da MMP-9 e os parâmetros: subtipo histológico, estádio, presença de sintomas B, idade e sexo. Não houve influência da MMP-9 na SLE (P = 0,98) e SG (P = 0,60). Conclusões: Demonstramos que a prevalência do LH relacionado ao EBV na população estudada é de 52,5%, e que a presença do vírus não altera a evolução clínica, SLE e SG de pacientes tratados uniformemente. Concluímos ainda que a MMP-9 é fortemente expressa nas células HRS. Não há correlação entre a expressão de MMP-9 e o status do EBV. Nem a SG nem a SLE foram influenciadas pela expressão dessa enzima. / Clinical and histological features of classical Hodgkin lymphoma (cHL) are primarily due to the effects of cytokines, enzymes and chemokines produced by Hodgkin-Reed-Sternberg (HRS) cells and their surrounding inflammatory cells in response to signals triggered by etiological factors such as Epstein-Barr virus (EBV). Matrix metalloproteinase-9 (MMP-9) has been associated with poorer survival in patients with aggressive non-Hodgkin lymphomas. In EBV-related cancers the expression of viral latent membrane protein 1 (LMP1) correlates with an increased MMP-9 expression. In this study, we evaluated the prognostic relevance of MMP-9 expression and EBV status in HRS cells in patients with cHL in Brazil. Material and Methods: We selected 97 patients with cHL. Patients were included if they had: 1) > 18 years, 2) Undergone similar chemotherapy protocols, 3) Paraffin blocks available for review and for EBV and MMP-9 detection and 4) Clinical, epidemiological and laboratorial parameters available. Results: EBV was detected in 52.5% of all cases. MMP-9 expression positivity was found in 87.6% of all cases. There was no correlation between MMP-9 expression and EBV status. Response to treatment and relapse rate were independent of MMP-9 expression and EBV status. When stratified according to chemotherapy protocol used or disease stage, we still did not find any difference. MMP-9 positivity did not influence overall survival and event free survival. Conclusion: MMP-9 are expressed in the majority of HRS cells and did not correlated with EBV status or survival. The consistent MMP-9 expression in HRS cells makes this enzyme a potential target for therapy. / TEDE / BV UNIFESP: Teses e dissertações

Introdução: Aneurisma de aorta é considerado uma doença inflamatória crônica, mas ainda é controverso se está relacionado a aterosclerose aos agentes infecciosos. Antígenos de Chlamydophila pneumoniae (CP) e Mycoplasma pneumoniae (MP) foram encontrados em grande quantidade nas placas instáveis que usualmente estão associadas a remodelamento positivo e inflamação do vaso. Metaloproteinase da matriz 9 (MMP-9) está implicada na fragilidade da parede vascular e na formação dos aneurismas. Os efeitos imunomodulatórios dos receptores ativados por proliferador de peroxissomo (PPARs) têm sido relacionados à aterosclerose. Objetivos: Comparar lesões ateroscleróticas graves com e sem aneurisma do ponto de vista inflamatório e infeccioso, analisando antígenos de MP e CP e expressão de MMP-9, TIMP-1 e PPARs. Métodos: No presente estudo quantificamos, através da técnica de imunoistoquímica, antígenos de MMP- 9, TIMP-1, PPARs a e , e dos agentes infecciosos CP e MP em fragmentos de aorta ateroscleróticos com aneurisma (n=14) e sem aneurisma (n=14). Resultados: A adventícia e o tecido adiposo periadventicial (TAP) dos aneurismas apresentaram intensa inflamação. Expressão de MMP-9 esteve aumentada no TAP e agentes infecciosos, TIMP-1 e PPAR a estiveram aumentados na adventícia e no TAP, sem diferença em relação a expressão de PPAR . Colágeno, TIMP-1 e PPARs estiveram positivamente correlacionados no grupo com aterosclerose, mas não no grupo com aneurisma. Conclusão: Nossos achados sugerem que aneurisma de aorta é uma complicação das lesões ateroscleróticas. Quantidade aumentada de Chlamydophila pneumoniae e Mycoplasma pneumoniae na adventícia e PAT sem correlação da resposta de TIMP-1 e PPARs são achados que estão associados com a presença de aneurisma nos segmentos de aorta ateroscleróticos / Introduction: Aortic aneurysm is considered a chronic inflammatory disease, but remains a controversial matter if it is related to atherosclerosis and infectious agents. Chlamydophila pneumoniae (CP) and Mycoplasma pneumoniae (MP) antigens were found in higher amount in unstable plaques that are usually associated to positive remodeling and vessel inflammation. Matrix metalloproteinase 9 (MMP-9) has been implicated in vascular wall fragility and aneurysm development. The immunemodulator effects of peroxisome proliferators - activated receptor (PPARs) have been related to atherosclerosis. Objectives: To compare severe atherosclerotic lesions with and without aneurysms by inflammatory and infectious point of view, analyzing MP and CP, MMP-9, TIMP-1 and PPARs antigens. Methods: In the present study we quantified, using immunohistochemistry technique, MMP-9, TIMP-1, PPAR a and and infectious agents CP and MP antigens in aortic atherosclerotic fragments with aneurysms. (n=14) and without aneurysms (n=14). Results: Adventitia and periadventitial adipose tissue (PAT) from aneurysms showed intense inflammation. MMP-9 expression has increased in PAT and the infectious agents, TIMP-1 and PPAR a were increased in adventitia and PAT in aneurysmatic group, without difference related to PPAR expression. Collagen, TIMP-1 and PPARs were positively correlated in atherosclerotic group, but it was not observed in aneurysmatic group. Conclusion: Our data have suggested that aortic aneurysm is a complication of aortic atherosclerotic lesions. Increased amount of Chlamydophila pneumoniae and Mycoplasma pneumoniae in the adventitia and PAT without a correlated TIMP-1 and PPARs response are findings that were associated with the presence of aneurysm in atherosclerotic aortic segments