A proteólise de componentes do plasma e da matriz extracelular é um fator importante no quadro hemorrágico desencadeado pela ação de venenos de serpentes. Neste estudo, avaliamos o papel dos domínios estruturais de metaloproteinases na interação com seus alvos. Ensaios de hidrólise in vitro mostraram que as metaloproteinases HF3 e bothropasina (classe P-III), e BJ-PI (classe P-I) do veneno da Bothrops jararaca, degradaram proteínas plasmáticas (fibrinogênio, fibronectina e vitronectina), além de colágeno VI e Matrigel, mostrando diferentes perfis de hidrólise. Experimentos de N-deglicosilação das proteinases mostraram que suas porções glicosídicas são importantes para a manutenção da integridade de suas estruturas e da atividade proteolítica. E ensaios para avaliar a interação das proteinases e da proteína DC (domínios tipo disintegrina/rico em cisteínas) com alguns substratos protéicos mostraram que além de seus domínios estruturais (catalítico/tipo disintegrina/rico em cisteínas) suas cadeias de carboidratos também são importantes no reconhecimento dos mesmos. / Proteolysis of plasma and extracellular matrix proteins is a key factor in the local hemorrhage induced by snake venom metalloproteinases (SVMPs). In this work we evaluated the role of the structural domains of SVMPs in the interaction with their targets. In vitro hydrolysis assays showed that the SVMPs HF3 and bothropasin (P-III class) and BJ-PI (P-I class) from Bothrops jararaca venom degraded plasma proteins (fibrinogen, fibronectin and vitronectin), collagen VI and Matrigel resulting in different hydrolysis profiles. N-deglycosylation of these proteinases showed that their carbohydrate moieties are important for keeping their structural integrity and proteolytic activity. The assays to analyze the interaction of the proteinases and of the DC protein (composed of the disintegrin-like and cysteine-rich domains) with some proteins confirmed that not only the structural domains (catalytic, disintegrin-like and cysteine-rich) but also the sugar moieties are important in the recognition of plasma and extracellular matrix proteins.

Orientador: Paulo Sérgio Cerri / Resumo: A periodontite (DP) é uma doença imunoinflamatória que promove o recrutamento de células específicas que liberam mediadores, dentre eles interleucina-1 (IL-1), fator de necrose tumoral-alfa (TNF-α) e metaloproteinases da matriz (MMPs). Há evidências de que a diacereína, uma medicação anti-IL-1, reduz a produção de MMPs e TNF-α. O objetivo do presente estudo foi avaliar se a diacereína acelera a regressão da inflamação e reduz a perda óssea na DP induzida. No total, foram utilizados 66 ratos Holtzman distribuídos, aleatoriamente, nos diferentes grupos. Em 42 ratos, a DP foi induzida com inserção de uma ligadura no 1º molar superior direito por 7 dias. Após a indução, 12 animais foram sacrificados: 6 animais do grupo GP (grupo periodontite) e 6 animais do grupo controle (GC), usado como parâmetro da DP induzida. No 7º dia de indução da DP, a ligadura colocada em 36 ratos foi removida e iniciou-se o tratamento com 100 mg/kg de massa corpórea de diacereína (GPD) ou solução fisiológica (GPS) por gavagem durante 7, 15 e 30 dias. Após 24 horas da administração da última dose, 6 animais de cada grupo (GPD, GPS e GC) foram sacrificados. Após eutanásia, fragmentos de maxila com molares do lado direito foram removidos e logo imersos em solução de formaldeído 4%. Em seguida, os espécimes foram descalcificados em EDTA 7% e processados para inclusão em parafina. De cada fragmento, 5 cortes sagitais semi-seriados foram corados com hematoxilina e eosina (HE) para análises morfológica e quant... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

Stimulation of β-adrenergic receptors (β-AR) induces apoptosis in adult rat ventricular myocytes (ARVMs) via the JNK-dependent activation of mitochondrial death pathway. Recently, we have shown that inhibition of matrix metalloproteinase-2 (MMP-2) inhibits β-AR-stimulated apoptosis and that the apoptotic effects of MMP-2 are possibly mediated via its interaction with β1 integrins. Herein we tested the hypothesis that MMP-2 impairs β1 integrin-mediated survival signals, such as activation of focal adhesion kinase (FAK), and activates the JNK-dependent mitochondrial death pathway. Inhibition of MMP-2 using SB3CT, a selective gelatinase inhibitor, significantly increased FAK phosphorylation (Tyr-397 and Tyr-576). TIMP-2, tissue inhibitor of MMP-2, produced a similar increase in FAK phosphorylation, whereas treatment of ARVMs with purified active MMP-2 significantly inhibited FAK phosphorylation. Inhibition of MMP-2 using SB3CT inhibited β-AR-stimulated activation of JNKs and levels of cytosolic cytochrome c. Treatment of ARVMs with purified MMP-2 increased cytosolic cytochrome c release. Furthermore, inhibition of MMP-2 using SB3CT and TIMP-2 attenuated β-AR-stimulated decreases in mitochondrial membrane potential. Overexpression of β1 integrins using adenoviruses expressing the human β1A-integrin decreased β-AR-stimulated cytochrome c release and apoptosis. Overexpression of β1 integrins also inhibited apoptosis induced by purified active MMP-2. These data suggest that MMP-2 interferes with the β1 integrin survival signals and activates JNK-dependent mitochondrial death pathway leading to apoptosis.

Indiana University-Purdue University Indianapolis (IUPUI) / Introduction: The resin-dentin interface undergoes degradation by endogenous matrix metalloproteinases (MMPs) after adhesive procedures. Application of several MMP inhibitors such as chlorhexidine (CHX) to the demineralized collagen dentin matrix after acid-etching has been suggested to be a successful approach to prevent degradation of the hybrid layer. Further, nanotubes (HNT) have been used as a reservoir for encapsulation and controlled delivery for several therapeutic drugs with sustained release. Therefore, HNT can be encapsulated with CHX and incorporated into dentin adhesives for the possibility of enhancing the longevity and durability of the hybrid layer. Objective: To evaluate the effect of a CHX-encapsulated nanotube-modified primer/PR and adhesive/ADH on the microtensile resin bond strength (µTBS) to dentin. Materials and Methods: A commercial adhesive and its respective primer were modified by adding CHX-encapsulated nanotubes at two distinct concentrations (10 and 20 wt.%). The experimental adhesives were evaluated by degree of conversion (DC) and viscosity. Meanwhile, only viscosity was determined for the experimental primers. The prepared HNT-encapsulated with CHX (10 and 20 wt.%) powders were incorporated into the primer and/or adhesive according to the groups: ADH (control); HNT (control); 0.2% CHX; PR+CHX10%; PR+CHX20%; ADH+CHX10%; ADH+CHX20%. Human molars were selected and autoclaved; mid-coronal dentin surfaces were exposed for bonding purposes. Dentin surfaces were etched, followed by primer and adhesive application, and restored with a resin composite. After 24 hours, the teeth were sliced into beams for µTBS testing; beams collected for each tooth were equally assigned into two testing condition groups: 24 hours and 6 months. Microtensile bond strength was tested using a universal testing machine, and the types of failure were classified as adhesive, mixed, and cohesive failure. Data from DC and viscosity tests were analyzed using one-way ANOVA. Bond strength data were analyzed by pair-wise comparisons using the Sidak method to control the overall significance level at 5% for each aging time separately. Weibull-distribution survival analysis was used to compare the differences in the microtensile bond strength results among the groups after 24 hours and 6 months. Results and Conclusion: DC analysis revealed no significant differences among adhesive groups. However, ADH group had a significantly lower viscosity than modified adhesive groups, and a significantly higher viscosity than modified primer groups. Test results of stress value (MPa) by each group for each aging time revealed no significant differences among groups after 24 hours. However, after 6-month storage, modified primer groups (PR+CHX10%, PR+CHX20%) and 0.2%CHX group showed a significant difference in µTBS compared to control groups (ADH, HNT) and modified adhesive groups (ADH+CHX10%, ADH+CHX20%) in the same aging time testing (p < 0.05). When comparing the µTBS after 24 hours and 6 months, there were no significant differences among the groups except for the ADH+CHX20% group, for which MPa values were higher after 24 hours than 6 months (p = 0.0487). In conclusion, this study has demonstrated the great potential of modified dental primers with CHX-encapsulated nanotubes in preservation of the resin-dentin bond strength over a 6-month time period. Additionally, modification of dental primers and adhesives was a successful approach that didn’t compromise the characteristics or the mechanical properties of the materials and has a promising long-term effect on resin-dentin bond strength.

Matrix metalloproteinases (MMPs) are a family of zinc endopeptidases capable of digesting the extracellular matrix (ECM), which is essential for tissue structure and transmitting messages between cells. MMPs play an important role in cancer, controlling cell migration, proliferation, apoptosis, regulation of tumour expansion, angiogenesis and invasion. Previous research has indicated high expression of MT1-MMP in breast cancers suggesting a potential role in tumour progression. Our results confirm that 3D multicellular tumour spheroids (MCTS) using phenotype-specific breast cancer cell lines are a valuable experimental model of the tumour microenvironment. Optimisation of MCTS culture growth conditions using different breast cancer cell lines (MCF-7, T47D, MDA-MB-468 and MDA-MB-231) was performed. Unexpected detection of MT1-MMP in MCF-7 MCTS warranted further investigation. MT1-MMP expression in different micro-environmental conditions, including hypoxia and nutrient deprivation (serum-free induced autophagy) were measured in MCF-7 monolayer cultures and MCTS models using immunofluorescence (IF), immunohistochemistry (IHC) and western blot (WB). MT1-MMP expression was rapidly and irreversibly up-regulated in MCF-7 breast cancer cells under conditions of stress (hypoxia and autophagy) compared to normal conditions suggesting an important role of the culture environment on cells behaviour and protein expression. We employed isobaric tags for relative and absolute quantitation (iTRAQ) technology to correlate MT1-MMP increase with proteomic profiles in MCF-7 breast cancer cell grown under hypoxic, serum-free and 3D MCTS conditions. More than 3500 proteins were identified, which were clustered into groups based on response to unique or shared microenvironment changes. Hypoxic monolayer and spheroid cells exhibited changes in anaerobic metabolism and lipid synthesis, respectively, whereas autophagy resulted in up-regulation of cellular component disassembly. The result indicated multiple drivers of MT1-MMP expression in MCF-7 cells. / Al-Mstansiriya University, Iraq

No description available.

Bone Cancer Research Trust / The full text will be available at the end of the embargo: 6th October 2027

No / Improved understanding of the involvement of matrix metalloproteinases (MMPs), including membrane-type MMPs (MT-MMPs), in human tumours has potential diagnostic, prognostic and therapeutic implications. We assessed the relationship between MT-MMP expression and clinicopathological parameters in human non-small cell lung cancer (NSCLC) and histologically normal lung tissue by quantitative Real Time PCR (qRT-PCR). All MT-MMPs (MMPs 14-17, 24 and 25) were detected by qRT-PCR with significantly higher MMP-14, -15 and -17 expression observed in tumour relative to normal lung specimens. MMP-16 was undetectable in normal lung but expressed in 8% tumours. MMP-15 demonstrated significant overexpression in adenocarcinomas relative to squamous cell carcinomas and normal lung tissue. MMP-14 mRNA expression strongly correlated to MMP-14 proteolytic activity in preclinical tumour models, indicating that qRT-PCR may predict MMP-14 activity levels in NSCLC. These data suggest that MMP-14, -15 and -17 may be good markers of disease, or therapeutic targets for treatment of human NSCLC.

No / Vascular disrupting agents (VDA) offer a strategy to starve solid tumors of nutrients and oxygen concomitant with tumor shrinkage. Several VDAs have progressed into early clinical trials, but their therapeutic value seems to be compromised by systemic toxicity. In this report, we describe the design and characterization of a novel VDA, ICT2588, that is nontoxic until activated specifically in the tumor by membrane-type 1 matrix metalloproteinase (MT1-MMP). HT1080 cancer cells expressing MT1-MMP were selectively chemosensitive to ICT2588, whereas MCF7 cells that did not express MT1-MMP were nonresponsive. Preferential hydrolysis of ICT2588 to its active metabolite (ICT2552) was observed in tumor homogenates of HT1080 relative to MCF7 homogenates, mouse plasma, and liver homogenate. ICT2588 activation was inhibited by the MMP inhibitor ilomastat. In HT1080 tumor-bearing mice, ICT2588 administration resulted in the formation of the active metabolite, diminution of tumor vasculature, and hemorrhagic necrosis of the tumor. The antitumor activity of ICT2588 was superior to its active metabolite, exhibiting reduced toxicity, improved therapeutic index, enhanced pharmacodynamic effect, and greater efficacy. Coadministration of ICT2588 with doxorubicin resulted in a significant antitumor response (22.6 d growth delay), which was superior to the administration of ICT2588 or doxorubicin as a single agent, including complete tumor regressions. Our findings support the clinical development of ICT2588, which achieves selective VDA targeting based on MT-MMP activation in the tumor microenvironment.

Les phosphates de calcium sont des biomatériaux couramment utilisés dans de nombreuses spécialités médicales. L'amélioration de ces biomatériaux vise à augmenter leur ostéointégration et leur bioactivité. Le strontium possédant d'intéressantes capacités de modification de la physiologie osseuse, l'incorporation de ce dernier au sein de phosphates de calcium par substitution ionique pourrait permettre un déplacement de la balance osseuse vers la formation osseuse.Notre travail a permis de démontrer la capacité des particules de phosphates de calcium substitués en strontium à augmenter la prolifération des ostéoblastes en culture et à modifier l'expression et la synthèse des principales protéines impliquées dans la physiologie osseuse (Collagène de type I, Serpine H1, métalloprotéinases matricielles 1 et 2, inhibiteurs tissulaires des MMPs). Par ailleurs, la poudre de phosphates de calcium ne contenant pas de strontium a entrainé une sécrétion accrue de chimiokines pro-inflammatoires (MCP-1 et GRO-?) qui n'a pas été observée pour la poudre substituée. Enfin, des études in-vivo réalisées dans un modèle de défaut osseux murin a permis de démontrer une plus grande résorbabilité de la poudre contenant du strontium et sa plus grande capacité à stimuler la réparation osseuse. / Calcium phosphate are widely used in medicine. Their upgrade tend to enhance their biocompatibility and their bioactivity. Strontium has interesting capability to modify the bone physiologie. Its incorporation in calcium phosphates could lead to modify the bone balance toward osteogenesis.The present work reveal the capacities of such biomaterials to enhance the replication of osteoblasts ant to modify the expression and the synthesis of proteins implicated in the bone balance (type I collagen, serpinH1, Matrix metalloproteinases 1 and 2, tissular inhibitors of MMPs). Moreover, non substituted calcium phosphate powders enhance the expression and synthesis of inflammatory cytokines (MCP-1 and Gro-a). This fact was not observed with the non substituted powder. In-vivo studies on a mouse model permit us to demonstrate the higher resorbability and the higher bone healing capability of the substituted powder.