Made available in DSpace on 2016-06-02T20:18:12Z (GMT). No. of bitstreams: 1 2857.pdf: 11303199 bytes, checksum: e1e51cdf6acc5b01445639c49dd0e72d (MD5) Previous issue date: 2010-03-17 / Universidade Federal de Minas Gerais / Peripheral nerves are frequently target of traumatic injuries and their functional recovery is generally incomplete. The aim of the present study was to evaluate the effects of 660 or 780 nm low-level laser therapy (LLLT) GaAlAs using different energy densities (10, 60 and 120 J/cm2) on nerve sciatic recovery after severe (neurotmesis) or moderate (axonotmesis) injuries in rat. One hundred and twenty eight Wistar male rats (275g) were divided into 16 groups, and they were performed as follow: For axonotmesis: Normal (N); axonotmesis (CR); axonotmesis and 660 nm LLLT with 10 J/cm2 irradiation (CR660 10J); CR660 60J; CR660 120J; CR780 10J; CR780 60J; CR780 120J. For neurotmesis: N; neurotmesis (TT); TT660 10J; TT660 60J; TT660 120J; TT780 10J; TT780 60J; TT780 120J. The LLLT irradiation was performed using a fix potence of 40 mW and a spot area of 4 mm2. Nerves submitted to axonotmesis or neurotmesis were irradiated with LLLT daily during 10 consecutive days starting on the first post-operatory. However, neurotmesis groups received additionally one month of LLLT applied every other day. Tibialis anterior (TA) muscles and sciatic nerve were evaluated 28 (axonotmesis) and 84 (neurotmesis) after surgery. The follow analyses were performed: muscle fiber, axon, myelin and nerve fiber cross-sectional area (CSA); matrix metalloproteinases (MMP) 2 and 9 activities; sciatic functional index; S-100 immunofluorescence. Data were submitted to homogeneity and normality tests. Anova one-way followed by Tukey tests and Kruskal-Wallis followed by Newman-Keuls tests were performed when data was parametric or non-parametric, respectively. Significance level was set at 5%. The results of the present study showed that groups irradiated with 660 nm LLLT with 10 or 60 J/cm2, and 780 nm 10 or 120 J/cm2 showed normal values of nerve fiber and myelin CSA. The 660 nm LLLT, regardless the energy density used, accelerated muscle fiber recovery and increased the MMP-2 activity in nerve. Furthermore, it also decreased the MMP-9 and MMP-2 activities in nerves and muscles respectively. All axotomized animals recovery normal levels of function on the 28 day after surgery. Regarding neurotmesis groups, TT660 120J presented higher values of myelin and nerve fiber CSA compared to TT. Superior values of muscle fiber CSA were observed in TT660 60 e 120J e TT780 10J compared to TT. These LLLT parameters were also efficient to decrease MMP-2 activity in TA muscles. All groups submitted to neurotmesis did not recover normal function after 84 days of injury. Based on the proposed objective, it is possible to conclude that LLLT, considering specific protocols vi of application, recovered nerves effectively, avoided muscle fiber atrophy and acted on the muscle and nerve extracellular matrix remodeling via MMPs regulation. / O objetivo do estudo foi analisar a ação dos lasers de baixa intensidade (GaAlAs) com diferentes comprimentos de ondas (660 e 780 nm), em diferentes densidades de energia (10 J/cm², 60 J/cm² e 120 J/cm2) na regeneração do nervo ciático do rato após lesões moderadas (axoniotmese) e graves (neurotmese). Para tal, 128 ratos Wistar (275 g) foram divididos em 16 grupos: Para axoniotmese: Normal (N); axoniotmese (CR); axoniotmese irradiado com laser 660 nm com 10 J/cm2 (CR660 10J); CR660 60J; CR660 120J; CR780 10J; CR780 60J; CR780 120J. Para neurotmese: N; neurotmese (TT); TT660 10J; TT660 60J; TT660 120J; TT780 10J; TT780 60J; TT780 120J. A estimulação a laser foi realizada com potência fixa de 40 mW e área do feixe de 4,0 mm2. O laser foi aplicado durante 10 dias consecutivos, a partir do primeiro dia do pós-operatório nos nervos com axoniotmese e neurotmese. Entretanto, para os grupos neurotmese foi acrescido de mais um mês de aplicação em dias alternados. Os músculos tibiais anteriores (TA) e os nervos ciáticos foram avaliados após 28 (axoniotmese) e 84 dias (neurotmese) após a cirurgia. Para investigar a área de secção transversa (AST) das fibras musculares, axônios, mielina e fibra nervosa; número de fibras nervosas foi feita morfometria. A atividade das metaloproteinases de matriz (MMPs) 2 e 9 foi realizada para observar o remodelamento da matriz. O índice funcional do nervo ciático foi para avaliar a função. A imunoflorescência com anticorpo S-100 foi utilizada para a observar a bainha de mielina e células de Schwann. Os dados foram submetidos a testes de homogeneidade e normalidade. Os testes Anova one-way seguido por Tukey e Kruskal-Wallis seguido por Newman-Keuls foram usados em amostras paramétricas e não paramétricas, respectivamente. Um nível de significância de 5% foi adotado. Os resultados deste estudo mostram que os grupos axoniotmese irradiados com lasers 660 nm 10 e 60 J/cm2, 780 nm 10 e 120 J/cm2 apresentaram AST da fibra nervosa e da mielina semelhante aos valores normais. O laser 660 nm, independente da densidade de energia utilizada, acelerou a recuperação da atrofia muscular e aumentou a atividade da MMP-2 nos nervos ciáticos dos grupos axoniotmese. Ele também reduziu a atividade da MMP-9 nos nervos e da MMP-2 nos músculos. Todos os animais submetidos à axoniotmese recuperaram a função normal após 28 dias de lesão. Em relação aos grupos neurotmese, foram observados valores superiores de AST da bainha de mielina e de fibra nervosa no grupo TT660 120J comparado ao TT. Valores de AST das fibras musculares foram superiores ao TT nos grupos TT660 60 e 120J e TT780 10J. Estes parâmetros de laser também foram eficientes em promover a diminuição da atividade de MMP-2 nos músculos TA. Todos os grupos submetidos à neurotmese não recuperaram totalmente a função após 84 dias de lesão. Com base no objetivo proposto podemos concluir que a laserterapia, em determinados protocolos de aplicação, mostrou-se efetiva na recuperação do nervo, evitou a atrofia do muscular e atuou no remodelamento da matriz extracelular do músculo e do nervo periférico via regulação das MMPs.

Made available in DSpace on 2016-06-02T19:22:03Z (GMT). No. of bitstreams: 1 2425.pdf: 1867201 bytes, checksum: 5e3f6810f116a13f8397936acba4c138 (MD5) Previous issue date: 2009-06-09 / Universidade Federal de Sao Carlos / As metaloproteases de matriz (MMPs) são cruciais para manutenção do tecido saudável. O objetivo deste estudo foi investigar a atividade da MMP-2 nos músculos gastrocnêmio, sóleo, tibial anterior (TA) e extensor longo dos dedos (EDL) após treinamento de força em ratas ovariectomizadas. Ratas Wistar adultas foram alocadas nos grupos: sedentário (Sed-Intacto); sedentário ovariectomizado (Sed-Ovx); sedentário pseudo-ovariectomizado (Sed-Pseudo); exercício agudo (AgudoEx-Intacto); exercício agudo ovariectomizado (AgudoEx-Ovx); treinamento de força (CrônicoEx-Intacto) e treinamento de força ovariectomizado (CrônicoEx-Ovx) (n= 10 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. A atividade da MMP-2 foi analisada por zimografia. Houve uma maior atividade da MMP-2 nos grupos CrônicoEx-Intacto e CrônicoEx-Ovx e menor atividade no AgudoEx-Ovx comparado com o grupo Sed-Intacto no sóleo (p≤0,05). Os grupos Sed-Ovx e CrônicoEx-Ovx apresentaram menor atividade da MMP-2 comparado com o grupo Sed-Intacto no TA. Houve maior atividade da MMP-2 no AgudoEx-Intacto e AgudoEx-Ovx comparado com Sed-Intacto e Sed-Ovx no TA, respectivamente (p≤0,05). Nos músculos TA e EDL o treinamento aumentou a atividade da MMP-2 comparado com o grupo Sed-Intacto. Não foram observadas alterações estatisticamente significativas para o músculo gastrocnêmio. O treinamento de força aumenta a atividade da MMP-2 nos músculos sóleo, TA e EDL, o que pode ser importante para o remodelamento muscular. A ovariectomia reduz a atividade da MMP-2 no TA e EDL, possivelmente comprometendo a função muscular.

Submitted by Aelson Maciera (aelsoncm@terra.com.br) on 2017-03-29T20:16:13Z No. of bitstreams: 1 DissNSMS.pdf: 2315984 bytes, checksum: 1245c9af0e2575904b71a6d6569edf15 (MD5) / Approved for entry into archive by Ronildo Prado (ronisp@ufscar.br) on 2017-04-11T19:16:23Z (GMT) No. of bitstreams: 1 DissNSMS.pdf: 2315984 bytes, checksum: 1245c9af0e2575904b71a6d6569edf15 (MD5) / Approved for entry into archive by Ronildo Prado (ronisp@ufscar.br) on 2017-04-11T19:16:36Z (GMT) No. of bitstreams: 1 DissNSMS.pdf: 2315984 bytes, checksum: 1245c9af0e2575904b71a6d6569edf15 (MD5) / Made available in DSpace on 2017-04-11T19:43:37Z (GMT). No. of bitstreams: 1 DissNSMS.pdf: 2315984 bytes, checksum: 1245c9af0e2575904b71a6d6569edf15 (MD5) Previous issue date: 2016-04-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Cervical cancer is the second largest cause of deaths from malignant cancer among women worldwide. Many studies show the importance of the HPV virus in the onset of cervical cancer, but because of the monoclonal nature of this cancer, it is suspected that HPV infection may not be considered the only causal factor responsible for the development of cervical carcinoma. Studies have indicated that EBV may have a role in cooperation with the HPV16 tumor development. Such cooperation could influence the tumor microenvironment by creating a favorable niche for its progression. Cancer cells secrete enzymes that degrade the extracellular matrix (ECM). Such enzymes such as matrix metalloproteinases (MMPs) facilitate cell movement in tissues and induce the activity of growth factors that promote angiogenesis. Among the group of MMPs, MMP-7 seems to have a crucial role in the development of carcinogenesis, since its action has been associated with the cleavage and release of adhesion molecules important. In this sense, this study was performed to establish the relationship between the HPV coinfection / EBV and expression of MMP-7. They were examined immunohistochemically histological slides of cervical lesions biopsies of 60 patients aged 35 to 65 years. The samples were divided into high grade lesion and low grade and evaluated semiquantitatively by scoring system ranging from 0 to +++. Histopathological indexes the degree of injury were established. It observed a statistically significant association between high-grade lesion, the expression of MMP-7 and the presence of the HPV virus, with p = 0.007 at the junction portion squamocolumnar and p = 0.023 in the portion of the ectocervix. / O câncer cervical é responsável pela segunda maior causa de mortes por neoplasia malignas entre as mulheres no mundo. Muitos estudos comprovam a importância do vírus HPV no surgimento do câncer cervical, porém devido à natureza monoclonal desta neoplasia, suspeita-se que a infecção pelo HPV não possa ser considerada o único fator causal responsável pelo desenvolvimento do carcinoma de colo de útero. Estudos já apontaram que o EBV pode ter um papel de cooperação com o HPV16 no desenvolvimento tumoral. Tal cooperação poderia influenciar o microambiente tumoral criando um nicho favorável para a sua progressão. Células de câncer secretam enzimas que degradam a matriz extracelular (MEC). Tais enzimas, como as metaloproteinases de matriz (MMPs) facilitam a movimentação celular nos tecidos e induzem a atividade de fatores de crescimento que promovem a angiogênese. Dentre o grupo das MMPs, a MMP-7 parece ter papel crucial no desenvolvimento da carcinogênese, pois sua ação já foi associada com a clivagem e a liberação de importantes moléculas de adesão. Neste sentido, foi realizado este estudo para estabelecer a relação entre a co-infecção HPV/EBV e a expressão da MMP-7. Foram examinadas imunohistoquimicamente lâminas histológicas de biópsias de lesões de colo uterino de 60 pacientes com idades entre 35 e 65 anos. As amostras foram divididas em lesão de alto grau e de baixo grau e avaliados semiquantitativamente através de um sistema de escore variando de 0 à +++. Índices histopatológicos quanto ao grau das lesões foram estabelecidos. Foi observada uma associação estatisticamente significante entre a lesão de alto grau, a expressão da MMP-7 e a presença do vírus HPV, com p=0,007 na porção da junção escamocolunar e p=0,023 na porção da ectocérvice.

Skeletal metastasis is a lethal component of many advanced cancers including prostate, the second most common cancer among men. Patients whose prostate cancer is localized and detected early benefit from multiple treatment options ranging from active surveillance to radiation and surgery, resulting in a 5-year survival rate of nearly 100%. Unfortunately, the prognosis and survival for patients with advanced metastatic disease is much worse due to the highly aggressive nature of the disease and a paucity of treatment options. Understanding the mechanisms and interactions that occur between metastatic cancer cells and the bone will enable the future treatment landscape for bone metastatic prostate cancer to expand, thereby improving patient outcomes. Our current knowledge of how metastatic prostate cancer cells interact with the bone is summarized in a model known as the “vicious cycle.” Numerous fundamental vicious cycle factors have been identified, including parathyroid hormone-related protein (PTHrP), while additional elements, such as matrix metalloproteinases (MMPs), are progressively being discovered and added to the model. PTHrP is a critical regulator of bone resorption and augments osteolysis in skeletal malignancies. In Chapter 2, we report that the mature PTHrP1-36 hormone is processed by MMPs to yield a stable product, PTHrP1-17. PTHrP1-17 retains the ability to signal through PTH1R to induce calcium flux and ERK phosphorylation but not cyclic AMP production or CREB phosphorylation. Notably, PTHrP1-17 promotes osteoblast migration and mineralization in vitro, and systemic administration of PTHrP1-17 augments ectopic bone formation in vivo. Further, in contrast to PTHrP1-36, PTHrP1-17 does not affect osteoclast formation/function in vitro or in vivo. Finally, immunoprecipitation-mass spectrometry analyses using PTHrP1-17-specific antibodies establish that PTHrP1-17 is indeed generated by cancer cells. Thus, MMP-directed processing of PTHrP disables the osteolytic functions of the mature hormone to promote osteogenesis, indicating important roles for this mechanism in bone remodeling in normal and disease contexts. MMPs have traditionally been associated with cancer progression based on their extracellular matrix degrading activities. However, it has become evident that their regulation of non-extracellular matrix substrates can exert both contributive and protective effects during tumorigenesis. Previous studies of matrix metalloproteinase-3 (MMP-3) have demonstrated tissue dependent pro- and anti-tumorigenic effects, but despite elevated expression, its roles have not been explored in bone metastatic prostate cancer. In Chapter 3, we show that tumor-derived MMP-3 contributes to prostate tumor growth in bone. In vitro, we observe that silencing MMP-3 reduces prostate cancer cell proliferation. Further, we found increased levels of IGFBP3, a known MMP-3 substrate, and decreased IGF-1R, ERK, and AKT phosphorylation in the MMP-3 silenced cells. Notably, we also observe reduced tumor growth and proliferation in in vivo intratibial models when tumor-derived MMP-3 expression is silenced. These data suggest that increased MMP-3 expression by prostate cancer cells contributes to their proliferation in bone by regulating the activity of the IGF/IGF-1R signaling axis. Taken together, our studies indicate that MMPs possess important functional roles in bone metastatic prostate cancer. We believe that elucidation of these mechanisms and their contributions to the vicious cycle of bone metastasis will offer novel opportunities to design effective therapeutic treatment options.

Abstract Breast carcinoma is the most common malignancy in females in western countries. Classical prognostic factors such as the size of a primary tumor and the presence or absence of axillary lymph node metastases, malignancy grade and hormone receptor status reflect the subsequent risk of disease recurrence after primary therapy and the need for adjuvant therapies. However, most breast carcinomas are detected in the early stage of the disease and the value of these classical prognostic factors is limited. There is also a great need to find new factors predicting the clinical efficacy of the anticancer drugs available. In this thesis tumor suppressor gene p53, oncogene c-erbB-2 and matrix metalloproteinase-9 were evaluated for their prognostic relevance in breast carcinoma patients treated in Oulu University Hospital, and matrix metalloproteinase-9 was also analyzed in women with premalignant lesions in the breast tissue in order to examine its role in breast carcinogenesis. Histological analyses were carried out from formalin-fixed, paraffin-embedded primary tumor specimens and p53, c-erbB-2 and matrix metalloproteinase-9 (MMP-9) statuses were systematically analyzed by immunohistochemistry. P53 expression correlated with disease-free survival and overall survival in patients with early-stage breast carcinoma, regardless of adjuvant antiestrogen therapy. The co-expression of p53 and c-erbB-2 characterizes a tumor type with a clinically aggressive course of breast carcinoma. The clinical efficacy of anthracyline-based chemotherapy in metastatic carcinoma might be limited in patients with p53 expression in a primary tumor. When postmenopausal patients with lymph node metastases and receiving adjuvant antiestrogen therapy were examined, MMP-9 expression indicated a slightly greater risk of breast carcinoma recurrence in patients with estrogen receptor negative tumors. Hyperplastic breast tissue and invasive breast carcinoma lesions expressed some MMP-9 immunopositivity. However, the strongest positivity was seen in ductal carcinoma in situ samples, suggesting that MMP-9 participates in breast carcinogenesis in the preinvasive phase.

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.

Abstract Dentin formation in physiological and pathological conditions has been widely studied, but the events and regulation are still not completely understood. Odontoblasts, terminally differentiated post-mitotic cells located in a single cell layer around pulp tissue, synthesize and mineralize dentin organic matrix. Growth factors, such as TGF-β1 and BMP-2, have been implicated in the regulation of the responses of odontoblasts and pulp tissue to external irritation. Matrix metalloproteinases (MMPs), a family of 28 endopeptidases collectively capable of degrading virtually all extracellular matrix components, and their specific tissue inhibitors (TIMPs) participate in the organo- and morphogenesis, physiological tissue turnover and pathological tissue destruction in many tissues, but very little is known about their presence, function, and regulation in the dentin-pulp complex cells and tissues. The aim of the work presented in this thesis was to analyze the expression and regulation of collagens, MMPs and TIMPs by TGF-β1 and BMP-2 in mature human odontoblasts and pulp tissue. Odontoblasts synthesize and secrete type I and type III collagens, with no clear effect of TGF-β1 on their expression levels. MMP-1, -2, -8, -9, -10, -11, -14, -15, -16, -19 and TIMP-1, -2, -3 and -4 were expressed by both odontoblasts and pulp tissue. MMP-3 and MMP-12 were not expressed in native odontoblasts or pulp tissue, and MMP-7, -24, and -25 were expressed only in odontoblasts. MMP-2, -10, -14, -20 and -23 were expressed more abundantly in odontoblasts, whereas pulp tissue expressed more MMP-13 and MMP-17. Growth factors differentially regulated the expression of different MMPs and TIMPs within and among the cells and tissues studied. In odontoblasts, MMP-1, -8 and -14 were down-regulated, but MMP-7, MMP-9, TIMP-1 and TIMP-3 up-regulated, by either TGF-β1 or BMP-2, alone or in combination. In pulp tissue, growth factors up-regulated the expression of MMP-1, -2, -10, -13, -17 and TIMP-3, but down-regulated TIMP-4. The widespread of expression of MMPs and TIMPs by mature human odontoblasts and pulp tissue suggests that they may participate in dentin matrix organization prior to mineralization, and that growth factors may further control dentin matrix modeling, not by regulating the synthesis of type I or III collagens as previously believed, but rather by differentially regulating each MMPs and TIMPs.

Abstract Breast carcinoma is a heterogeneous disease with a prognosis that varies from excellent to very poor. Traditional tumour parameters and biological factors that are also predictive for treatment response are used in determining breast carcinoma prognosis and selecting appropriate treatment. Gelatinases MMP-2 and MMP-9 have been shown to associate with tumour progression. Their tissue inhibitors TIMP-1 and -2 are multifunctional molecules that have been suggested as prognostic markers in some previous reports. In the present work, the expression and prognostic value of gelatinases MMP-2 and MMP-9 and their tissue inhibitors TIMP-1 and -2 were assessed in primary breast carcinoma. The material consisted of a total of 416 patients. Tissue expression of TIMP-1 and -2 was analysed in a population of 203 patients using immunohistochemistry. Circulating gelatinases and their inhibitors were studied using ELISA in two different populations of 71 at preoperative state and 213 patients at pre- and postoperative state. High expression of TIMP-1 immunoreactive protein positively correlated with high histological grade of the tumour and associated with aggressive disease course in grade 2–3 subpopulation. High preoperative plasma TIMP-1 was prognostic for relapse in a modern patient series after a median follow-up time of 18 months. TIMP-1 as a continuous variable was prognostic in Cox regression univariate analysis, and was an independent prognostic variable superior to nodal status in multivariate analysis. High preoperative serum TIMP-1 was an independent prognostic variable for poor disease-specific survival, and TIMP-1 was found to maintain its prognostic value when assessed independently with different ELISA analyses, and was not very sensitive for preanalytical conditions. In addition, low circulating preoperative serum MMP-2 was observed to associate with high stage and positive nodal status in breast carcinoma. These results indicate that circulating TIMP-1 may be a potential new marker of worsened prognosis in breast carcinoma, although careful validation of assay platforms and identification of the sources of physiological variation are needed before it can be adopted into clinical decision-making.

Naegleria fowleri is a free-living amoeba found in freshwater lakes and ponds that is the causative agent of Primary Amoebic Meningoencephalitis (PAM). Matrix metalloproteinases (MMPs) have been described in protozoa, such as Plasmodium falciparum, Trypanosoma brucei, and Balamuthia mandrillaris, and have been linked to their increased motility and invasive capability by degrading components of the extracellular matrix (ECM). In addition, MMPs are often upregulated in tumorigenic cells and have been attributed as responsible for the metastasis of certain cancers. In the present study, in vitro experiments indicated that MMPs are linked functionally to the ECM degradation process. Gelatin zymography demonstrated protease activity in N. fowleri whole cell lysates, conditioned media, and media collected from in vitro invasion assays. Western immunoblotting confirmed the presence of the metalloproteinases MMP-2, -9, and -14. The highly virulent mouse-passaged amoebae expressed higher levels of MMPs than the weakly virulent axenically grown amoebae. The functional relevance of MMPs found in media in degradation of ECM components was confirmed through the use of MMP inhibitors. The collective in vitro results suggest that MMPs may play a critical role in the invasion of the CNS. Furthermore, the expression of select metalloproteinases may serve as amenable targets for therapeutic manipulation of expansive PAM.

Cutaneous wound healing could be distinguished into two main types: embryonic and adult. Embryonic healing in contrast to adult is faster, scar-less and consists of early, middle, and late phases. Actin ring is formed during the early phase and its cables pull the edges of the wound towards apposite sites during the following middle phase. De novo expression of healing specific genes is initiated also during middle phase. However, process of the wound healing continues under the closed wound in the late phase which has been poorly described. Adult wound healing is more complex, longer, and is divided into 4 phases: haemostasis, inflammation, proliferation, and remodelling phase. Adult wound healing might end with the scar. Pivotal role in the wound healing is given to matrix metalloproteinases (MMPs). These remodelling enzymes are important for releasing cytokines, inducing apoptosis, and degradation of extracellular matrix. Our laboratory performed temporal RNA-sequencing of the healing tissue using tailbud stage and swimming tadpole embryos. Results showed predominant expression of four mmps: mmp1.L, mmp7.S, mmp8.S, and mmp9.L. Injury or amputation caused the upregulation and their expression level peaked at 3-6 hours post injury which corresponds with late phase of healing. Thus, I focused on...