Αν και η Αυχενική Σπονδυλωτική Μυελοπάθεια (ΑΣΜ) αποτελεί την πιο κοινή αιτία δυσλειτουργίας νωτιαίου μυελού στους ενήλικες άνω των 55 ετών, οι μοριακοί μηχανισμοί παραμένουν άγνωστοι. Μέχρι σήμερα, πολλές προσπάθειες έχουν διενεργηθεί για την ανάπτυξη ενός αξιόπιστου πειραματικού μοντέλου AΣΜ. Ωστόσο, αρκετά μειονεκτήματα εμφανίζονται σε αυτές τις μελέτες. Στη παρούσα μελέτη έχουμε σκοπό τη δημιουργία ενός νέου, πρωτότυπου πειραματικού μοντέλου ΑΣΜ, το οποίο εξομοιώνει τα ιστολογικά και κλινικά χαρακτηριστικά της ανθρωπίνης νόσου. Mεθοδολογία: Μετά από αφαίερεση του πετάλου του έβδομου αυχενικού σπονδύλου, ένα λεπτό τεμάχιο αρωματικού πολυαιθέρα τοποθετήθηκε κάτω από το πέταλο του έκτου αυχενκού σπονδύλου σε κόνικλους Νέας Ζηλανδίας (Ομάδα ΧΠΠ). Σε μία άλλη ομάδα πειραματόζωων ο αρωματικός πολυαιθέρας αφαιρέθηκε 30 δευτερόλεπτα μετά την εμφύτευση (ομάδα ελέγχου). Νευρολογική εκτίμηση πραγματοποιήθηκε χρησιμοποιώντας τη κλίμακα του Tarlov μετά το πέρας της χειρουργικής διαδικασίας και ακολούθως εβδομαδιαίως. Ηλεκτροφυσιολογικές μελέτες πραγματοποιήθηκαν στις 20 εβδομάδες μετά το χειρουργείο και πριν από τη θυσία των πειραματόζωων. Ακολούθησαν ιστολογικές και ανοσοιστοχημικές μελέτες. Αποτελέσματα: Τα πειραματόζωα που άνηκαν στην ομάδα ελέγχου δεν εμφάνισαν νευρολογικά ελλείμματα κατά τη διάρκεια της μελέτης. Αντιθέτως τα πειραματόζωα που άνηκαν στη ΧΠΠ εμφάνισαν νευρολογικά ελλείματα. Στους νωτιαίους μυελούς προερχόμενους από την ΧΠΠ ομάδα ανεδείχθησαν οι χαρακτηριστικές ιστοπαθολογικές αλλοιώσεις της χρόνιας μυελοπάθειας. Ειδικότερα, ανεδείχθη σπογγώδης εκφύλιση της λευκής ουσίας, διάμεσο οίδημα και αποπλάτυνση των πρόσθιων κεράτων της φαιάς ουσίας. Επίσης ανεδείχθη κατακρήμνιση του μυελικού σάκου και διόγκωση του δακτυλίου της μυελίνης. Τέλος, η χρόνια πίεση του νωτιαίου οδήγησε σε ενεργοποίηση της απόπτωσης και διαταραχή της αρχιτεκτονικής του μικροαγγειακού συστήματος του νωτιαίου μυελού Συμπέρασμα: Το πρωτότυπο μοντέλο ΑΣΜ στους κονίκλους ποσομοιώνει το χωρικό και χρονικό προφίλ της ανθρώπινης νόσου στο σημείο της πίεσης του νωτιαίου μυελού. ΜΕΛΕΤΗ B Εισαγωγή: Η φλεγμονή, η δημιουργία ουλώδους ιστού και η διαταραχή του μικροαγγειακού συστήματος του νωτιαίου μυελού είναι ορισμένα από τα κύρια παθοφυσιολογικά φαινόμενα της ΑΣΜ. Ωστόσο οι μοριακοί μηχανισμοί που εμπλέκονται σε αυτά τα φαινόμενα κάτω από τη χρόνια και προοδευτική πίεση του νωτιαίου μυελού παραμένουν ανεξερεύνητα. Mεθοδολογία: Στη συγκεκριμένη μελέτη χρησιμοποιήθηκε το πειραματικό μοντέλο ΑΣΜ που περιγράφεται στη μελέτη Α με σκοπό να διερευνηθεί ο ρόλος του NF-κB και των πρωτεινών της εξωκυττάριας ουσίας στην ΑΣΜ. Εν συντομία, κόνικλοι Νέας Ζηλανδίας (διαφορετικά πειραματόζωα από εκείνα της μελέτης Α) χωρίστηκαν τυχαία σε δύο ομάδες: την ομάδα ΧΠΠ (n=15) και την ομάδα ελέγχου (n=15). Η έκφραση των πρωτεινών των υπομονάδων p50 και p65 του NF-kB, όπως επίσης και των ενζύμων διάσπασης της εξωκυττάριας ουσίας (MMP-2, MMP-9) και του ενεργοποιητή του πλασμινογόνου τύπου ουροκινάσης (urokinase-type plasminogen activator; u-PA) αξιολογήθηκαν σε τομές νωτιαίων μυελών προερχόμενων και από τις δύο ομάδες χρησιμοποιώντας ανοσοιστοχημική τεχνική. Στατιστική ανάλυση πραγματοποιήθηκε χρησιμοποιώντας SPSS για Windows, release 12.0 (SPSS Inc., Chicago, IL). Αποτελέσματα: Σε τομές νωτιαίων μυελών που προέρχονταν από πειραματόζωα που έπασχαν από ΑΣΜ αναδείχθηκε στατιστικά σημαντικά αυξημένη έκφραση των υπομονάδων του NF-κB (p50 & p65), όπως επίσης και των ενζύμων MMP-2, MMP-9, and u-PA σε σύγκριση με εκείνες που προέρχονταν από την ομάδα ελέγχου. Τέλος, σημαντικά θετική συσχέτιση παρατηρήθηκε μεταξύ των επιπέδων έκφρασης του NF-κB και εκείνων των MMP-9, MMP-2, and u-PA. Συμπέρασμα: Τα ευρήματα αυτά αποτελούν ισχυρές ενδείξεις πως η χρόνια και προοδευτική πίεση του αυχενικού νωτιαίου μυελού οδηγεί σε αυξημένη έκφραση των MMP-2, MMP-9 και u-PA πιθανόν μέσω της δράσης του μεταγραφικού παράγοντα NF-κB. Είναι βέβαιο ότι περισσότερες μελέτες απαιτούνται για την εξακρίβωση του ρόλου των πρωτεινών αυτών στην ΑΣΜ. / Although cervical spondylotic myelopathy (CSM) represents the most common cause of spinal cord impairment among individuals over 55 years old, the molecular mechanisms of the disease remain mainly unknown. To date, many experimental studies have been conducted to establish a reliable model of CSM, however most of them appear some limitations. In this study we aim to create a new animal model of CSM, which will reproduce the temporal course of the human disease and the local microenvironment at the site of spinal cord compression. Methods: Following C7 posterior laminectomy, a thin sheet of aromatic polyether was implanted underneath C5–C6 laminae of the New Zealand rabbits. A sham group in which the material was removed 30 sec after the implantation was also included. Motor function evaluation was performed after the material implantation and weekly thereafter using the Tarlov classification. At 20 weeks post-material implantation electrophysiological studies were also conducted. All the animals were sacrificed 20 weeks post-material implantation and histological and immunohistochemical studies were performed. Results: Clinical evaluation of animals after operation reveals no symptoms and signs of acute spinal cord injury. Moreover, no neurological deficits were noticed in the sham group during the course of the study. However, the animals which underwent implantation of compression material exhibited progressive neurological deficits throughout the study. Rabbits of the compression group experienced significant increased axonal swelling and demyelination, interstitial edema and myelin sheet fragmentation. Histological evaluation of C5 and C6 laminae (at the site of implantation) reveals osteophyte formation. Moreover, the chronic and progressive compression of the cervical spinal cord resulted in induction of apoptosis as well as in disruption of the basement membrane of vessels. Conclusion: The proposed rabbit CSM model reproduces the temporal evolution of the disease and creates a local microenvironment at the site of spinal cord compression, which shares similar features with that of human disease. STUDY B Introduction: Inflammation, glial scar formation and disruption of spinal cord microvasculature represent some of the principal neuropathological features of CSM. However, the molecular mechanisms which are implicated in these pathophysiological phenomena under the chronic and progressive compression of the cervical spinal cord remain interestingly unexplored. Methods: In this study (B) in order to evaluate the role of NF-κB and extracellular matrix proteins in cervical myelopathy we used the rabbit CSM model which was extensively characterized in study A. Briefly New Zealand rabbits (different cohort of animals than that of the study A) were randomly and blindly divided into the following two groups: CSM (n=15) and sham group (n=15). The expression pattern of p50 and p65 subunits of NF-kB, as well as that of MMP-2, MMP-9, and u-PA, was evaluated in spinal cord sections coming from both groups using immunohistochemistry technique. Statistical analysis was performed using SPSS for Windows, release 12.0 (SPSS Inc., Chicago, IL). Results: CSM animals exhibited statistically significant increased immuoreactivity in both NF-κB subunits, p50 and p65. Moreover, the levels MMP-2, MMP-9, and u-PA were found to be significantly increased in CSM animals compared to controls. Finally, strong positive correlation between NF-κB subunits immunoreactivity and that of MMP-9, MMP-2, and u-PA was demonstrated. Conclusion: The NF-κB pathway as well as the extracellular matrix proteins (MMP-2 and MMP-9) are involved in CSM. However, more studies are needed to clarify the functional role of these molecules in the pathobiology of CSM.

Made available in DSpace on 2014-06-11T19:32:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-03-27Bitstream added on 2014-06-13T20:22:59Z : No. of bitstreams: 1 kanno_cm_dr_araca.pdf: 1730975 bytes, checksum: 620b9c72f44529033f574688f3ea9bde (MD5) / INTRODUÇÃO: As alterações em gengiva induzidas por medicamentos têm sido pouco estudadas quanto à expressão in vivo dos genes das metoloproteinases (MMPs). O objetivo do presente trabalho foi avaliar o padrão histológico de distribuição de fibras colágenas após a administração de ciclosporina, nifedipina ou fenitoína e correlacionar com a expressão dos genes do colágeno do tipo I, MMP-1 e MMP2. MATERIAL E MÉTODO: Amostras da gengiva da área de canino superior direito foram obtidas de doze macacos prego (Cebus apella) machos. A extremidade mesial de cada amostra foi imediatamente congelada em nitrogênio líquido enquanto que a distal foi processada para inclusão em parafina. Após uma semana, os animais foram divididos em três grupos que receberam doses diárias de ciclosporina, fenitoína ou nifedipina, durante 120 dias. Procedeu-se à remoção de amostras da gengiva da área do canino superior esquerdo de dois animais de cada grupo aos 52 e 120 dias. Os cortes histológicos foram corados pelas técnicas da hematoxilina e eosina, vermelho picrosirius, além da marcação imunoistoquímica para colágeno do tipo IV. O RT-PCR semiquantitativo foi realizado para se determinar os níveis de mRNA. RESULTADOS: No grupo controle, houve o predomínio de fibras colágenas maduras, evidenciadas com a cor vermelha em cortes corados pela técnica do vermelho picrosirius analisados com microscópio de luz polarizada. Observou-se nos grupos tratados aos 52 e 120 dias um aumento da porcentagem de áreas ocupadas por fibras imaturas, em todos os grupos, independentemente da idade do animal. No entanto, não foram observadas diferenças morfológicas entre os grupos controle e tratado nos cortes corados pela hematoxilina e eosina. Houve uma tendência a valores médios mais baixos... / Background: Few studies have focused on the in vivo expression of matrix metalloproteinase (MMP) genes in gingival changes induced by drugs. The aim of the present study was to evaluate the histological pattern of collagen fiber distribution after phenytoin, cyclosporine or nifedipine medication and correlate with collagen type 1, MMP-1 and MMP-2 gene expression levels. Methods: Gingival samples were obtained from superior right canine area of twelve male capuchin monkeys (Cebus apella). The mesial part of the biopsy specimens was immediately frozen in liquid nitrogen, while the distal one was processed for paraffin inclusion. One week after the control biopsy, the animals were divided in three groups that received daily doses of cyclosporine, phenytoin or nifedipine during 120 days. Gingival samples were obtained from left superior canine area on 52nd and 120th day of treatment (two animal of each experimental group). Histologic sections were subjected to hematoxylin and eosin, picrosirius red stainings, and to immunohistochemical reaction for collagen type IV. MMP-1, MMP-2 and collagen type I mRNA levels were determined by RT-PCR. Results: Predominance of mature collagen fibers was observed in the control group after picrosirius red staining, visualized as red fibers under polarized microscope. Increased percentage of areas occupied by immature collagen fibers was observed on 52 and 120 experimental periods, in all groups, despite the animal age. However, no morphological differences between treated and control groups were observed on hematoxilin and eosin stained sections. There was a trend to lower levels of MMP-1 expression on 52-day samples. However, MMP-2 and collagen type I gene expressions seemed to be phased and drug-related. Conclusions: The results allowed the ...(Complete abstract click electronic access below)

Orientador: Alvimar Lima de Castro / Banca: Renata Tucci / Banca: José Fernando Garcia / Banca: Sérgio Roberto Peres Line / Banca: José Américo de Oliveira / Resumo: INTRODUÇÃO: As alterações em gengiva induzidas por medicamentos têm sido pouco estudadas quanto à expressão in vivo dos genes das metoloproteinases (MMPs). O objetivo do presente trabalho foi avaliar o padrão histológico de distribuição de fibras colágenas após a administração de ciclosporina, nifedipina ou fenitoína e correlacionar com a expressão dos genes do colágeno do tipo I, MMP-1 e MMP2. MATERIAL E MÉTODO: Amostras da gengiva da área de canino superior direito foram obtidas de doze macacos prego (Cebus apella) machos. A extremidade mesial de cada amostra foi imediatamente congelada em nitrogênio líquido enquanto que a distal foi processada para inclusão em parafina. Após uma semana, os animais foram divididos em três grupos que receberam doses diárias de ciclosporina, fenitoína ou nifedipina, durante 120 dias. Procedeu-se à remoção de amostras da gengiva da área do canino superior esquerdo de dois animais de cada grupo aos 52 e 120 dias. Os cortes histológicos foram corados pelas técnicas da hematoxilina e eosina, vermelho picrosirius, além da marcação imunoistoquímica para colágeno do tipo IV. O RT-PCR semiquantitativo foi realizado para se determinar os níveis de mRNA. RESULTADOS: No grupo controle, houve o predomínio de fibras colágenas maduras, evidenciadas com a cor vermelha em cortes corados pela técnica do vermelho picrosirius analisados com microscópio de luz polarizada. Observou-se nos grupos tratados aos 52 e 120 dias um aumento da porcentagem de áreas ocupadas por fibras imaturas, em todos os grupos, independentemente da idade do animal. No entanto, não foram observadas diferenças morfológicas entre os grupos controle e tratado nos cortes corados pela hematoxilina e eosina. Houve uma tendência a valores médios mais baixos ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Background: Few studies have focused on the in vivo expression of matrix metalloproteinase (MMP) genes in gingival changes induced by drugs. The aim of the present study was to evaluate the histological pattern of collagen fiber distribution after phenytoin, cyclosporine or nifedipine medication and correlate with collagen type 1, MMP-1 and MMP-2 gene expression levels. Methods: Gingival samples were obtained from superior right canine area of twelve male capuchin monkeys (Cebus apella). The mesial part of the biopsy specimens was immediately frozen in liquid nitrogen, while the distal one was processed for paraffin inclusion. One week after the control biopsy, the animals were divided in three groups that received daily doses of cyclosporine, phenytoin or nifedipine during 120 days. Gingival samples were obtained from left superior canine area on 52nd and 120th day of treatment (two animal of each experimental group). Histologic sections were subjected to hematoxylin and eosin, picrosirius red stainings, and to immunohistochemical reaction for collagen type IV. MMP-1, MMP-2 and collagen type I mRNA levels were determined by RT-PCR. Results: Predominance of mature collagen fibers was observed in the control group after picrosirius red staining, visualized as red fibers under polarized microscope. Increased percentage of areas occupied by immature collagen fibers was observed on 52 and 120 experimental periods, in all groups, despite the animal age. However, no morphological differences between treated and control groups were observed on hematoxilin and eosin stained sections. There was a trend to lower levels of MMP-1 expression on 52-day samples. However, MMP-2 and collagen type I gene expressions seemed to be phased and drug-related. Conclusions: The results allowed the ...(Complete abstract click electronic access below) / Doutor

Made available in DSpace on 2016-06-02T19:22:05Z (GMT). No. of bitstreams: 1 3128.pdf: 3115695 bytes, checksum: a3cd2a9e5289c0b15425f8203b8ca0c1 (MD5) Previous issue date: 2010-07-21 / INTRODUÇÃO: osteoporose é reconhecida mundialmente como sério problema de saúde pública, apresentando como característica principal a fragilidade óssea. A qualidade do osso é fator determinante nesse aspecto, e depende do colágeno. Um dos fatores intrínsecos que regula o colágeno é a atividade da metaloproteinase -2 (MMP-2). O treinamento de força é o melhor recomendado para preservar e/ou melhorar a qualidade da massa óssea. OBJETIVO: foi investigar a influência do treinamento de força na atividade da MMP-2 e nas propriedades mecânicas do osso de ratas ovariectomizadas (OVX) e intactas. MATERIAIS E MÉTODOS: 48 ratas maduras jovens foram distribuídas em 2 grupos distintos, ovariectomia (OVX) e Intactas (Int); a seguir, 3 subgrupos foram formados similarmente em cada grupo: sedentário (OVX-Sed e Int-Sed), exercício agudo (OVX-Ex-Ag e Int-Ex-Ag) e exercício crônico (OVX-Ex-Cr e Int-Ex-Cr) (n = 8 por grupo). Foi utilizado um treinamento de força de 12 semanas no qual os animais escalaram uma escada vertical de 1,1-m com pesos presos as suas caudas. As sessões foram realizadas com intervalo de três dias, 4-9 escaladas e 8-12 movimentos dinâmicos por escalada. Após o término do período experimental, foram realizadas análises da atividade da MMP-2 por zimografia e análises biomecânicas e biofísicas utilizando-se uma máquina de ensaio universal (Instron modelo 4444). RESULTADOS: a atividade da MMP-2 apresentou-se reduzida em 2 grupos OVX (OVXSed e OVX-Ex-Ag) comparada com todos os outros grupos (p ≤ 0,05). Em contrapartida, os grupos treinados cronicamente (OVX-Ex-Cr e Int-Ex-Cr) apresentaram aumento significativo da MMP-2. Esses resultados também foram observados nas análises biomecânicas e biofísicas, nas quais os grupos OVX-Sed e OVX-Ex-Ag apresentaram Densidade Mineral, Densidade Óssea, Carga Máxima e Carga de Fratura menores em relação à todos os outros grupos; na outra mão, os grupos treinados cronicamente, apresentaram índices maiores nas análises biomecânicas e biofísicas acima citadas. CONCLUSÕES: a ovariectomia reduziu a atividade da MMP-2, produzindo efeitos deletérios sobre a massa óssea. O treinamento de força proposto foi eficiente em combater esses efeitos, apresentando inclusive efeito modelador.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-06-03T23:19:48Z No. of bitstreams: 1 GeorgeAlexandreLira_DISSERT.pdf: 12487121 bytes, checksum: e5befba62b31cf9c0849235dd847b32c (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-06-06T23:43:09Z (GMT) No. of bitstreams: 1 GeorgeAlexandreLira_DISSERT.pdf: 12487121 bytes, checksum: e5befba62b31cf9c0849235dd847b32c (MD5) / Made available in DSpace on 2016-06-06T23:43:09Z (GMT). No. of bitstreams: 1 GeorgeAlexandreLira_DISSERT.pdf: 12487121 bytes, checksum: e5befba62b31cf9c0849235dd847b32c (MD5) Previous issue date: 2015-08-31 / Excluindo-se os tumores de pele n?o-melanoma, o c?ncer colorretal ? o segundo mais comum no sudeste do Brasil; o terceiro na regi?o sul e na regi?o Centro-Oeste. J? no norte do Brasil, ? o quarto e, na regi?o Nordeste, o quinto. Avaliar vari?veis clinico-patol?gicos do c?ncer colorretal ? de fundamental import?ncia para se conhecer poss?veis desfechos na sobrevida dos pacientes portadores e pontuar caracter?sticas na progress?o tumoral como o perfil da invas?o tumoral e angiog?nese. O objetivo desse trabalho ? estudar as caracter?sticas cl?nico-patol?gicas dos pacientes portadores do c?ncer colorretal (CCR) na Liga Norte Riograndense contra o c?ncer em Natal-RN/BR, analisando as vari?veis cl?nicas e patol?gicas como par?metros de progn?stico e determinando o n?vel de express?o de prote?nas, tais sejam: E-caderina (E-cad), Beta-catenina (?-cat), Galectina-3 (Gal-3), Metaloproteinases de matriz (MMP) 2 e 9 e o Fator alfa de crescimento v?sculo-endotelial (VEGF-?) nos tecidos tumorais. Foi realizado um estudo retrospectivo dos casos de c?ncer colorretal da Liga Norte-Riograndense contra o C?ncer no per?odo de 1995 a 2005 em Natal-RN / Brasil. As vari?veis cl?nico-patol?gicas, tais como: idade, sexo, etnia, h?bitos de vida, hist?ria familiar, local do tumor prim?rio, grau de diferencia??o, estadiamento TNM, Dukes? modificado, tratamento e sobrevida foram analisadas. Os dados cl?nico-patol?gicos foram coletados dos prontu?rios m?dicos atrav?s de um formul?rio espec?fico e os dados foram armazenados em uma planilha do Excel. Um total de 534 pacientes foi selecionado do arquivo do setor da patologia dessa institui??o, mas 176 pacientes foram exclu?dos. 358 pacientes foram inclu?dos para an?lise epidemiol?gica e suas correla??es cl?nico-patol?gicas foram realizadas. 180 pacientes foram selecionados para estudos histol?gicos e imunohistoqu?micos. Prote?nas participantes da progress?o tumoral E-caderina, Beta-catenina, Galectina-3, Metaloproteinases 2 e 9 e o Fator alfa de crescimento endotelial vascular foram analisadas. Os blocos de parafina dessas amostras foram tratados pela t?cnica de Tissue Microarray e suas l?minas submetidas a imunohistoqu?mica para avaliar a intensidade de marca??o dessas prote?nas nos tecidos tumorais. Os resultados dessa an?lise foram correlacionados ?s vari?veis cl?nico-patol?gicas dos pacientes. An?lise estat?stica pelo Teste de qui-quadro de Pearson e an?lise de sobrevida pela Curva de Kaplan-Meier foram utilizados. Valores de p<0,05 foram considerados estatisticamente significativos. A m?dia de idade da nossa amostra foi de 58,8 anos e 51,7% foram do sexo feminino. O consumo de ?lcool aumentou em 1,71 vezes o risco de morte pelo CCR (p=0,034). J? o tabaco aumentou 2,7 vezes a chance de desenvolver tumores de alto est?gio TNM (p=0,001). Os pacientes com hist?rico familiar de c?ncer teve 3,833 vezes a chance de desenvolver o CCR (p=0,002). A express?o da MMP-2 mostrou uma associa??o significativa com os tumores de alto est?gio TNM (p<0,046) e mortalidade (p=0,041). A express?o do ? VEGF teve correla??o estatisticamente significante com o alto est?gio TNM (p<0,009), grau de indiferencia??o celular (p<0,025) e mortalidade (p<0,035). As express?es da E-caderina e Beta-catetina mostraram associa??o do tumor com alto est?gio TNM (p=0,0001) e Dukes? modificado (p=0,05), les?o em reto (p=0,03 e p=0,007, respectivamente), tabaco (p=0,05) e indiferencia??o (p=0,001). A express?o das Gal-3 apresentou relev?ncia estat?stica com pacientes de alto est?gio TNM (p=0,01), fumantes (p=0,01), etilista (p=0,03), indiferencia??o (p=0,0001) e mortalidade (p=0,0001). Frente aos resultados, pode-se perceber que o estilo de vida e hist?rico familiar teve correla??o no progn?stico do CCR, assim como a express?o de MMP-2, MMP-9, VEGF alfa, E-caderina, Beta-catenina e Galectina-3 foram importantes marcadores de progn?stico na progress?o tumoral no c?ncer colorretal. / Except the non-melanoma skin tumors, colorectal cancer is the second most common in the Southeastern Region of Brazil, the third most common in the Southern and Central Regions. It is also the forth most common in the Northern Region and it is the fifth one in the Northeastern. To assess pathological and clinical variables of colorectal Cancer is crucial to know the possible conclusions for the survival of patients and point out the characteristics in the progress of tumor, such as the profile of tumor invasion and its angiogenesis. This work focuses on analyzing clinically and pathologically some settings in colorectal cancer patients (CRC) in the city of Natal and its countryside through those variables as parameters of prognosis and determine the level of protein expression, for instance: E-cadherin (E-cad), beta- -catenin (?-cat), galectin-3 (gal-3), matrix metalloproteinases (MMP) 2 and 9 and vascular-endothelial growth factor alpha (? VEGF) in the tumor tissues. A retrospective study was done in colorectal cancer cases in the regions of Rio Grande do Norte state from 1995 to 2005, specifically in Natal city/RN/Brazil. The pathological and clinical variables, such as: age, gender, ethnicity, lifestyle, family history, the location of the primary tumor, level of differentiation, TDM staging, modified Dukes?, treatment and survival were analyzed. The pathological and clinical data were collected from medical records through a specific form and were filed on Excel. A total of 534 patients were selected from the Pathology Department file in this institution, however, 176 patients were excluded. 358 patients were included for Epidemiological analysis and its clinical and pathological correlations were selected. 180 patients were also selected for histological and immunohistochemical studies. The tumor progression of these selected proteins mentioned before were analyzed. The Paraffin blocks of these samples were treated by Microarray Tissue technique and its blades subjected to immunohistochemistry to test the intensity of these proteins in tumor tissues. The results of this analysis were correlated with clinicopathologic variables of patients. Statistical analysis using the chi-frame Pearson test and analysis of midlife by Kaplan-Meier curve was also utilized. P values < 0.05 were considered statistically significant. The average age of our sample was 58.8 years and 51.7 % were female. Alcohol consumption has increased by 1.71 time the risk of death by CCR (p = 0.034) and tobacco consumption increased 2.7 times the chance of developing tumors of high TNM stage (p = 0.001). Cancer patients had a family history of 3,833 times the chance of developing the CCR (p = 0.002). The expression of MMP-2 showed a significant association with tumors of high TNM stage (p <0.046) and mortality (p = 0.041). The ? VEGF expression had statistically significant correlation with high TNM stage (p <0.009), degree of cell indifferentiation (p <0.025) and mortality (p <0.035). Expressions of E-cadherin and beta-catetina demonstrated tumor linked to high TNM stage (p = 0.0001) and Dukes? modified (p = 0.05), lesions in the rectum (p = 0.03 and p = 0.007, respectively), smoking (p = 0.05) and indifferentiation (p = 0.001). The expression of Gal-3 showed statistical significance with high TNM stage of patients (p = 0.01), smokers (p = 0.01), alcohol drinking (p = 0.03), indifferentiation (p = 0.0001) and mortality (p = 0.0001). Based on the results, therefore, we could realize that lifestyle and family history had correlation in the CCR prognosis, as well as MMP-2 expression, MMP-9, VEGF alpha, E-cadherin, Beta-catenin and Galectin-3 were important prognostic markers in tumor progression in colorectal cancer.

Abstract Endometrial carcinoma is the most common gynegologic malignancy in developed countries. Due to early symptoms, including abnormal uterine bleeding, endometrial cancer is often diagnosed at an early stage and in that case usually has a good prognosis and high cure rates. However, the nature of the disease is heterogeneous. During the last decades, the improvement in survival rates among endometrial cancer patients has not been significant, suggesting that the traditional clinicopathological factors may be inadequate to identify patients with high-risk disease. Furthermore, aggressive adjuvant treatments can be costly and very toxic. Therefore, better prognostic markers associated with biological aggressiveness of endometrial carcinoma are needed to identify the patients with high-risk disease, and to be able to select the treatment more individually. Gelatinases (MMP-2 and MMP-9) and their tissue inhibitors (TIMP-1 and TIMP-2) have been found to play a role in tumor progression. In the present work, the expression and prognostic value of MMP-2, MMP-9, TIMP-1 and TIMP-2 were assessed in endometrial carcinoma. The patient material consisted of a total of 266 women diagnosed with primary endometrial carcinoma. The tissue expression of immunoreactive proteins was examined in paraffin-embedded tumor sections by immunohistochemical staining using specific antibodies, and the pretreatment serum levels of the proteins were quantitatively measured by ELISA. Tissue MMP-2 expression associated with a worsened prognosis, whereas tissue TIMP-2 overexpression was an indicator of a favorable outcome. Furthermore, we observed a combination of strong MMP-2 and weak TIMP-2 tissue expression to identify a group of women at high risk of adverse outcome in endometrial carcinoma. Patients with negative MMP-2 immunostaining had the best prognosis, regardless of TIMP-2 staining result. In serum measurements, high preoperative TIMP-1 concentration was a prognostic indicator of unfavorable outcome. These results indicate that tissue MMP-2 and TIMP-2 as well as circulating TIMP-1 may be prognostic markers in endometrial carcinoma. Of these, tissue MMP-2 seems to be the most potent prognostic marker. Studies with larger patient materials are needed to further explore the value of these enzymes in clinical practice in endometrial cancer. / Tiivistelmä Kohdunrungon syöpä on yleisin gynekologinen maligniteetti kehittyneissä maissa. Varhaisten oireiden, kuten poikkeavan verisen vuodon, vuoksi kohdunrungon syöpä havaitaan usein varhaisessa vaiheessa, jolloin sen ennuste on hyvä. Taudin käyttäytyminen voi kuitenkin olla moninaista. Viime vuosikymmenten aikana kohdunrungon syöpään sairastuneiden ennuste ei ole merkittävästi parantunut. Vaikuttaisi siltä, että perinteiset ennustetekijät eivät ole riittävän tarkkoja ennustamaan syövän taudinkulkua. Lisäksi liitännäishoidot voivat olla kalliita, ja niihin voi liittyä vakavia haittavaikutuksia. Uusien biologisten ennustetekijöiden löytäminen olisi tärkeää, jotta aggressiivista syöpätyyppiä sairastavat potilaat pystyttäisiin tunnistamaan entistä paremmin, ja hoito kyettäisiin räätälöimään yksilöllisemmin taudinkuvaa vastaavasti. Gelatinaasien (MMP-2 ja MMP-9) sekä niiden kudosinhibiittoreiden (TIMP-1 ja TIMP-2) on havaittu osallistuvan syövän etenemiseen. Tässä tutkimuksessa tarkasteltiin MMP-2:n ja MMP-9:n sekä niiden kudosinhibiittoreiden TIMP-1:n ja TIMP-2:n ilmentymistä ja ennusteellista merkitystä kohdunrungon syövässä. Aineisto käsitti yhteensä 266 primaariseen kohdunrungon syöpään sairastunutta naista. Määritysmenetelminä käytettiin sekä immunohistokemiallista värjäystä parafiiniin valettujen kudosnäytteiden osalta että ELISA-määrityksiä ennen hoitoa otettujen seeruminäytteiden osalta. Syöpäkudoksen runsas MMP-2 -proteiinin ilmentyminen liittyi epäsuotuisaan ennusteeseen, kun taas kasvainkudoksen voimakas TIMP-2 -proteiinin ilmentyminen oli hyvän ennusteen merkki. Lisäksi kasvainkudoksen voimakkaan MMP-2- ja heikon TIMP-2 -proteiinien ilmentymisen yhdistelmän havaittiin liittyvän suurempaan syövästä johtuvaan kuolleisuuteen. MMP-2 -negatiivisten potilaiden eloonjäämisennuste oli paras, TIMP-2 -värjäystuloksesta riippumatta. Seerumin korkea TIMP-1 -pitoisuus oli merkittävä huonontuneen ennusteen merkki. Tutkimuksen tulokset viittaavat siihen, että kasvainkudoksessa esiintyvät MMP-2- ja TIMP-2 -proteiinit samoin kuin seerumin TIMP-1 -pitoisuus voivat ennustaa kohdunrungon syövän kliinistä käyttäytymistä. Kasvainkudoksessa esiintyvä MMP-2 -proteiini vaikuttaisi olevan merkittävin ennusteellinen tekijä, mutta tulosten varmistamiseksi tarvitaan lisää tutkimuksia suuremmilla potilasaineistoilla.

Εισαγωγή: Ο καρκίνος του λάρυγγα, ιδιαιτέρως σε προχωρημένα στάδια, είναι μία καταστροφική νόσος η οποία χαρακτηρίζεται από αυξημένη διηθητικότητα και μεταστατικότητα. Η ανεύρεση ενός δείκτη πρώιμης διάγνωσης, παρακολούθησης και πρόγνωσης της νόσου θα ήταν ιδιαίτερα ευπρόσδεκτη. Συνεχώς αυξανόμενα δεδομένα στη βιβλιογραφία υποστηρίζουν την προγνωστική αξία των ζελατινασών και τον πιθανό ρόλο τους ως μοριακών δεικτών μεταξύ άλλων και στον καρκίνο του λάρυγγα. Σκοπός: Η διαπίστωση μεταβολών στα επίπεδα ορού των ζελατινασών Α και Β σε ασθενείς με καρκίνο του λάρυγγα μετά από εφαρμογή θεραπείας, καθώς και η πιθανή συσχέτιση με διάφορες κλινικοπαθολογικές παραμέτρους πριν και μετά τη θεραπευτική παρέμβαση. Υλικό και μέθοδος: Σαράντα εννέα ασθενείς και 8 υγιείς μάρτυρες συμπεριλήφθηκαν στη μελέτη. Ελήφθησαν προεγχειρητικά και μετεγχειρητικά δείγματα ορού τα οποία στη συνέχεια υποβλήθηκαν σε ζυμογραφία ζελατίνης. Η παρουσία ζελατινασών επιβεβαιώθηκε με την τεχνική western blotting. Οι ζώνες λύσης ποσοτικοποιήθηκαν με τη χρήση Scion Image PC. Η ανάλυση των αποτελεσμάτων πραγματοποιήθηκε με το πρόγραμμα SPSS 17 (SPSS Inc, Chicago, IL, USA). Αποτελέσματα: Στα ζυμογραφήματα αποτυπώθηκαν μόνο οι λανθάνουσες μορφές των ενζύμων (προένζυμα). Τα προ της θεραπείας επίπεδα και των δύο ζελατινασών στον ορό του αίματος των ασθενών με καρκίνο του λάρυγγα ήταν σημαντικά υψηλότερα σε σχέση με αυτά των υγιών μαρτύρων. Ασθενείς με υπεργλωττιδικό καρκίνωμα και ενεργοί καπνιστές είχαν σημαντικά υψηλότερα επίπεδα proMMP-2 σε σχέση με ασθενείς που έπασχαν από γλωττιδικό καρκίνωμα και με πρώην καπνιστές αντίστοιχα. Ασθενείς με πρωτοδιαγνωσμένη νόσο και ασθενείς με λεμφαδενικές μεταστάσεις είχαν σημαντικά χαμηλότερα προ της θεραπείας επίπεδα proMMP-9 σε σχέση με ασθενείς που προσήλθαν με υποτροπή και με ασθενείς στους οποίους δεν διαπιστώθηκε επιχώρια νόσος αντίστοιχα. Κατά τη διάρκεια της συστηματικής παρακολούθησης τα επίπεδα της proMMP-2 στον ορό παρουσίασαν σημαντική αύξηση τις πρώτες 10 με 15 ημέρες μετά την εφαρμογή θεραπείας, για να μειωθούν σταδιακά εντός των επόμενων μηνών. Οι ενεργοί καπνιστές παρουσίασαν σημαντική μείωση των επιπέδων της proMMP-2 κατά την περίοδο παρακολούθησης, σε αντίθεση με τους πρώην καπνιστές οι οποίοι εμφάνισαν σημαντική αύξηση κατά το ίδιο χρονικό διάστημα. Οι ασθενείς σταδίου ΙΙ είχαν σημαντικά χαμηλότερα επίπεδα proMMP-2 σε σχέση με ασθενείς προχωρημένων σταδίων πέντε με οκτώ μήνες μετά τη θεραπεία, όπως και οι ασθενείς οι οποίοι υποβλήθηκαν σε συντηρητική αντιμετώπιση σε σχέση με τους χειρουργημένους ασθενείς. Τα επίπεδα της proMMP-9 στον ορό επίσης παρουσίασαν σημαντική πτώση μετά την εφαρμογή θεραπείας. Διαφορές στο ρυθμό μείωσης των επιπέδων της proMMP-9 παρατηρήθηκαν μεταξύ των διαφόρων ομάδων ως προς το στάδιο, τη διαφοροποίηση, την εντόπιση, τον τύπο της νόσου (πρωτοδιαγνωσμένη ή υποτροπή), τις λεμφαδενικές μεταστάσεις, τον τρόπο αντιμετώπισης και την κατανάλωση αλκοόλ. Ωστόσο αυτή η διαφορά δεν διατηρήθηκε πέντε με οκτώ μήνες μετά την εφαρμογή θεραπείας, με εξαίρεση την ομάδα των χειρουργημένων ασθενών, οι οποίοι διατήρησαν σημαντικά υψηλότερα επίπεδα ενζύμου στον ορό. Αύξηση των ζελατινασών παρατηρήθηκε στον ορό ασθενών που εκδήλωσαν υποτροπή μετά από αντιμετώπιση πρωτοδιαγνωσμένης νόσου σε σχέση με αυτούς που δεν υποτροπίασαν. Ωστόσο εξαιτίας του μικρού δείγματος δεν είναι δυνατόν να εξαχθούν ασφαλή συμπεράσματα. Συμπεράσματα: Αν και δεν υφίστανται φυσιολογικές τιμές, το πρότυπο μεταβολής των επιπέδων της proMMP-9 στον ορό μετά από θεραπεία καταδεικνύει πιθανές ιδιότητες μοριακού δείκτη. Ωστόσο υπάρχουν ενδείξεις ότι και οι δύο ζελατινάσες θα μπορούσαν να χρησιμοποιηθούν για την εξατομικευμένη παρακολούθηση ασθενών με καρκίνο του λάρυγγα. Περαιτέρω έρευνα απαιτείται για την αποσαφήνιση του ζητήματος. / Introduction: Laryngeal cancer, especially in the advanced stages, is a highly devastating disease, characterized by increased invasiveness and high rates of metastasis. The identification of reliable tumour marker for prompt diagnosis, surveillance and prognosis would be highly desirable. There is a growing body of evidence with regard to the prognostic value of gelatinases and their possible role as tumour markers. Aim: To identify the pattern of alteration of serum gelatinases A and B in patients with laryngeal cancer following treatment, and a possible correlation with various clinicopathological parameters prior to and past treatment. Materials and methods: Forty nine patients and 8 healthy controls were included in the study. Pre-treatment and post-treatment serum samples were collected and processed by gelatin zymography. The presence of gelatinases was verified by western blotting. The zymograms were scanned by a digital scanner and the lysis bands were quantified by Scion Image PC. Analysis of the quantitative results was performed by using SPSS 17 (SPSS Inc, Chicago, IL, USA). Results: Only the latent forms of MMP-2 and -9 (proforms) were identified. Both gelatinases were increased in the serum of laryngeal cancer patients compared to healthy individuals. Patients with supraglottic tumours and active smokers had significantly higher pre-treatment levels of proMMP-2 than patients with glottic tumours and ex-smokers, respectively. Patients with primary disease and patients with lymph node involvement showed lower proMMP-9 pre-treatment levels than patients with recurrence and patients without neck disease, respectively. During the follow-up period the proMMP-2 serum levels increased significantly in the first ten to fifteen days after treatment, gradually decreasing over the following months. Smokers showed a very high decrease rate of proMMP-2 levels during the follow-up period, whereas in ex-smokers proMMP-2 levels significantly increased. Stage II patients showed significantly lower levels of circulating enzyme compared to patients with more advanced disease five to eight months past treatment. Similarly, conservative management was associated with lower levels of serum proMMP-2 compared to surgical management five to eight months following treatment. The proMMP-9 serum levels also showed a gradual decrease after treatment, which was statistically significant. Significant alterations in the rate of decrease developed among groups with regard to stage, grade, location, type of disease (primary or recurrence), regional disease, treatment modality and alcohol consumption. Nevertheless those differences were not maintained five to eight months past treatment, with the exception of patients who underwent surgery and who maintained higher levels of proMMP-9. An increase to the levels of both gelatinases were observed in patients with recurrent disease after having been treated for a primary compared to patients who did not develop a recurrence. However, the small sample of patients with recurrent disease during the follow-up period does not allow extrapolating sound conclusions. Conclusions: Although as yet normal values have not been established in the literature, the post-treatment alteration pattern of proMMP-9 serum levels indicates that this enzyme might play a role as a tumour marker. Nevertheless this study provides evidence that both gelatinases might be useful for surveillance on strictly individual basis in laryngeal cancer patients. Further research is necessary to clarify the contribution of both gelatinases to the disease progress and determine their role as prognostic factors and tumour markers.

No description available.

Made available in DSpace on 2016-06-02T19:22:57Z (GMT). No. of bitstreams: 1 5061.pdf: 12379581 bytes, checksum: c5740c3bf0a93d60530d4f55769c222d (MD5) Previous issue date: 2013-03-22 / Universidade Federal de Sao Carlos / Obesity is associated with increased lipid peroxidation in the myocardium and an increased susceptibility to oxidative damage, which can lead to several cardiovascular risk factors including heart failure, insulin resistance, hypertension and dyslipidemia. The ONOOin conjunction with normal intracellular levels of reduced glutathione (GSH), are responsible, among other factors, to activate the matrix metalloproteinases (MMPs) that are important in mediating the deleterious effects of cardiovascular diseases. The unregulated activity of MMPs, such as MMP-2, may promote the development of heart disease. Resistance training (RT) can mitigate or even reverse the obesity, prevent against lipid peroxidation, increasing the levels of antioxidant enzymes, improved lipid profile, reduced cardiovascular risk and reduce the concentration of fibrosis and dysregulation of MMP-2. Purpose: To analyze the effects of RT high intensity on ladder associated with high fat diet on the activity of MMP-2 and antioxidant enzymes (SOD Total, Mn-SOD, CAT, GPx), GSH and lipid peroxidation, related to left ventricle remodeling in rats. Methods: 60 rats Wistar were used in this study, which was divided into three phases: 1) three week period of obesity induction, control group fed with standard diet (CT-S n = 30) and control group fed with high fat diet (CT-H n = 30); 2) effect of fat diet for eight weeks, sedentary standard diet group (SED-S n = 10) and high fat diet sedentary (SED-H n = 10), 3) effect of high-intensity RT on ladder for eight weeks associated with high-fat diet, RT standard diet (RT-S n = 10) and RT high-fat diet RT (RT-H n = 10). Results: The three-week obesity induction was effective in promoting the group CTH increased body mass (p <0.05), LV mass (p <0.05) and activity of MMP-2 (p <.001), TBRs (p <0.03) and decrease the citrate synthase activity (p <0.05). After eight weeks of high fat diet, the SED-H group had a higher body mass (p <0.02) compared to SED-S. The volume LV for SED-H group was higher compared to that of CT-H (p <0.001), but did not change when compared to SED-S. There was a lower activity of MMP-2 (p <0.05) in SED-S compared to CT-H. Lipid accumulation was observed (p <0.007) in the LV of SED-H group compared to CT-H. The longer life associated with high-fat diet promoted suppression in the enzymatic activity of SOD Total (p <0.01) and Mn-SOD (p <0.002) (SED-H), and increased activity of GSH (p <0.004). The RT was effective in attenuating the gain body mass in RT-H group compared to SED-H, even though higher compared to RT-S (p <0.005). In the group RT-H of MMP-2 activity was higher (p <0.03) compared to RT-S, but lower (p <0.04) compared to SED-H. The RT promoted greater citrate synthase activity (p <0.03), SOD Total (p <0.001) and Mn-SOD (p <0.03) and lower activity of GSH (p <0.01) in RT-H. There was no significant increase in TBARs level in the VE of the RT-H group. However, we observed increased lipid content (p <0.04). Conclusion: We conclude that high intensity RT, in obese rats, could be a pharmacological tool able to positively modulate the activity of MMP-2 and some antioxidant enzymes such as SOD Total, Mn-SOD and GSH. These effects are important for the maintenance of heart health, relieving the deleterious effects on obesityrelated of the left ventricle remodeling, so experiment. / A obesidade está associada com aumento da peroxidação lipídica no miocárdio e com uma maior suscetibilidade ao dano oxidativo, o que pode levar a vários fatores de risco cardiovascular incluindo insuficiência cardíaca, resistência à insulina, hipertensão e dislipidemia. O ONOO- em conjunto com níveis normais intracelular de glutationa reduzida (GSH), são responsáveis, dentre outros fatores, por ativar as metaloproteinases de matriz (MMPs), que são importantes na mediação dos efeitos deletérios de doenças cardiovasculares. A atividade desregulada das MMPs, como a MMP-2, pode favorecer o desenvolvimento de doenças cardíacas. O treinamento resistido (TR) pode amenizar ou mesmo reverter o quadro de obesidade, prevenir contra a peroxidação lipídica, aumentando os níveis das enzimas antioxidantes, melhora do perfil lipídico, diminui os riscos á doenças cardiovasculares, além de reduzir a concentração de fibrose e a desregulação da MMP-2. Objetivo: Analisar os efeitos do TR de alta intensidade em escada associado à dieta hiperlipídica na atividade da MMP-2 e enzimas antioxidantes (SOD Total, Mn-SOD, CAT, GPx), GSH e peroxidação lipídica, relacionadas ao remodelamento do ventrículo esquerdo, em ratos. Metodologia: 60 ratos Wistar foram utilizados neste estudo, sendo este divido em três fases: 1) período de três semanas de indução à obesidade, grupo controle alimentado com dieta padrão (CT-P n= 30) e controle alimentado com dieta hiperlipídica (CT-H n= 30); 2) efeito da dieta hiperlipídica por mais oito semanas, grupo sedentário dieta padrão (SED-P n= 10) e sedentário dieta hiperlipídica (SED-H n= 10); 3) efeito do TR em escada por oito semanas associado a dieta hiperlipídica, grupo TR dieta padrão (TR-P n= 10) e TR dieta hiperlipídica (TR-H n= 10). Resultados: As três semanas de indução à obesidade foi eficaz em promover aumento da massa corporal (p < 0,05), massa do VE (p < 0,05) e da atividade da MMP-2 (p < 0,001), do TBRs (p < 0,03) e de diminuir a atividade da citrato sintase (p < 0,05) no grupo CT-H. Após mais oito semanas de dieta hiperlipídica, o grupo SED-H teve a massa corporal maior (p < 0,02) comparado ao SED-P. Já a massa do VE para o grupo SED-H foi maior em comparação ao do CT-H (p < 0,001), porém não se modificou quando comparado ao SEP-P. Houve uma menor atividade da MMP-2 (p < 0,05) no grupo SED-H comparado ao CT-H. Foi observado acúmulo de lipídios totais (p < 0,007) no VE do grupo SD-H comparado ao CT-H. O maior tempo de vida associado à dieta hiperlipídica promoveu inibição da SOD Total (p < 0,01) e Mn-SOD (p < 0,002) (SED-H), e aumento da GSH (p < 0,004). O TR foi eficaz em atenuar o ganho de massa corporal no grupo TR-H comparado ao SED-H, mesmo sendo maior comparado ao TR-P (p < 0,005). No grupo TR-H a atividade da MMP-2 foi maior (p < 0,03) comparado ao TR-P, porém menor (p < 0,04) comparado ao SED-H. O TR promoveu maior atividade das enzimas citrato sintase (p < 0,03), SOD Total (p < 0,001) e Mn-SOD (p < 0,03) menor atividade da GSH (p < 0,01) no grupo TR-H. Não foi observado aumento significativo do TBARS no VE do TR-H. Contudo, foi observado aumento do conteúdo lipídico (p < 0,04). Conclusão: Concluímos que o TR de alta intensidade, em ratos obesos, pode ser uma ferramenta não farmacológica capaz de modular de forma positiva a atividade da MMP-2 e de algumas enzimas antioxidantes como a SOD Total, Mn-SOD e a GSH. Efeitos esses que são importantes para a manutenção da saúde cardíaca, amenizando os efeitos deletérios relacionados à obesidade sobre o remodelamento do ventrículo esquerdo, de forma experimental.