Alteration of the pharmacokinetics of COL-3, a matrix metalloproteinase inhibitor, due to actue gastrointestinal tosicity of doxorubicin

<b>Purpose </b>Combination of COL-3, a matrix metalloproteinase inhibitor, and doxorubicin (DOX) might be a promising anticancer regimen. The present study was to examine the potential pharmacokinetic interactions and toxicity profile following their coadministration in rats.<br /><b>Methods</b> Normal rats were treated with single agent or different combinations with oral or intravenous COL-3 and DOX, and the bile-duct cannulated (BDC) rats received oral COL-3 plus DOX. In a separate disposition study, the effects of DOX on the biliary, urinary, and fecal excretion of COL-3 were examined. In addition, the effects of DOX on in vitro protein binding, metabolism, and transport of COL-3 across Caco-2 monolayers were investigated.<br /><b>Results </b>COL-3 did not affect the pharmacokinetics of DOX in rats. However, treatment with DOX significantly decreased the oral absorption, and prolonged the elimination, of COL-3 in the normal rats, but not in the BDC rats. DOX did not alter the biliary and urinary excretion of COL-3, but significantly decreased the fecal excretion of COL-3. DOX significantly enhanced the basolateral to apical flux of COL-3 across Caco-2 monolayers, but had no apparent effects on the protein binding and metabolism of COL-3. The combination of DOX with oral COL-3 did not significantly (p &gt; 0.05) increase the acute diarrhea score and intestinal damage compared to rats receiving DOX alone.<br /><b>Conclusions</b> These results indicated that DOX altered the oral absorption and elimination of COL-3, largely resulting from gastrointestinal toxicity caused by biliary excretion of DOX. Further studies are required to explore the efficacy and optimized dosage regimen of this promising combination.<br /><br />

Matrix Metalloproteinase-9 reduces islet amyloid formation by degrading islet amyloid polypeptide

Deposition of islet amyloid polypeptide (IAPP) as amyloid is a pathological hallmark of the islet in type 2 diabetes, which is toxic to &beta;-cells. We previously showed that the enzyme neprilysin reduces islet amyloid deposition and thereby reduces &beta;-cell apoptosis, by inhibiting fibril formation. Two other enzymes, matrix metalloproteinase (MMP)-2 and MMP-9, are extracellular gelatinases capable of degrading another amyloidogenic peptide, A&beta;, the constituent of amyloid deposits in Alzheimer disease. We therefore investigated whether MMP-2 and MMP-9 play a role in reducing islet amyloid deposition. MMP-2 and MMP-9 mRNA were present in mouse islets but only MMP-9 activity was detectable. In an islet culture model where human IAPP (hIAPP) transgenic mouse islets develop amyloid but nontransgenic islets do not, a broad spectrum MMP inhibitor (GM6001) and an MMP-2/9 inhibitor increased amyloid formation and the resultant &beta;-cell apoptosis. In contrast, a specific MMP-2 inhibitor had no effect on either amyloid deposition or &beta;-cell apoptosis. Mass spectrometry demonstrated that MMP-9 degraded amyloidogenic hIAPP but not nonamyloidogenic mouse IAPP. Thus, MMP-9 constitutes an endogenous islet protease that limits islet amyloid deposition and its toxic effects via degradation of hIAPP. Because islet MMP-9 mRNA levels are decreased in type 2 diabetic subjects, islet MMP-9 activity may also be decreased in human type 2 diabetes, thereby contributing to increased islet amyloid deposition and &beta;-cell loss. Approaches to increase islet MMP-9 activity could reduce or prevent amyloid deposition and its toxic effects in type 2 diabetes.

Inhibition of matrix metalloproteinase activity in the chick sclera and its effect on myopia development

To investigate the contribution of matrix degradation in the two-layer avian sclera to the development of myopia. Tissue inhibitor of metalloproteinase-2 (TIMP-2) was used to inhibit chick scleral collagen degradation with (3)H-proline, a marker for this effect. Ex vivo scleral culture experiments confirmed TIMP-2 doses for in vivo experimentation. Ocular growth and refractive response to exogenous TIMP-2 (11.25, 2.25, and 0.45 picomoles, plus vehicle only) were monitored in 7-day-old chicks during the induction of myopia over 4 days with a translucent occluder. Collagen degradation was assessed, in whole sclera and in separated scleral layers by using the same paradigm (11.25 picomoles TIMP-2; vehicle only).Approximately 60% of collagen degradation was inhibited with low (2 nM) doses of TIMP-2 in the ex vivo sclera. Degradative activity in the in vivo chick sclera increased significantly (46%) during myopia development, with all the altered activity confined to the fibrous layer. Addition of TIMP-2 significantly reduced (by 46%) this accelerated scleral collagen degradation, also by acting in the fibrous layer. TIMP-2 had no significant effect on (3)H-proline incorporated in the cartilaginous scleral layer and cornea. Despite inhibiting collagen degradation TIMP-2 had no significant effect on myopia development. Increased collagen degradation is a feature of scleral remodeling in chick myopia development, but is confined to the fibrous scleral layer. Significant inhibition of this collagenolytic activity with TIMP-2 has little effect on refractive error development, suggesting that collagen degradation in the sclera contributes little to the development of myopia in the chick.

Characterization of the zebrafish matrix metalloproteinase 9 gene and its developmental expression pattern

Members of the matrix metalloproteinase (MMP) family are important for the remodeling of the extracellular matrix in a number of biological processes including a variety of immune responses. Two members of the family, MMP2 and MMP9, are highly expressed in specific myeloid cell populations in which they play a role in the innate immune response. To further expand the repertoire of molecular reagents available to study zebrafish myeloid cell development, the matrix metalloproteinase 9 (mmp9) gene from this organism has been identified and characterized. The encoded protein is 680 amino acids with high homology to known MMP9 proteins, particularly those of other teleost fish. Maternal transcripts of mmp9 are deposited in the oocyte and dispersed throughout the early embryo. These are replaced by specific zygotic transcripts in the notochord from 12 h post fertilization (hpf) and also transiently in the anterior mesoderm from 14 to 16 h post fertilization. From 24 h post fertilization, mmp9 expression was detected in a population of circulating white blood cells that are distinct from macrophages, and which migrate to the site of trauma, and so likely represent zebrafish heterophils. In the adult, mmp9 expression was most prominent in the splenic cords, a site occupied by mature myeloid cells in other species. These results suggest that mmp9 will serve as a useful marker of mature myeloid cells in the zebrafish.

Measurement of a MMP-2 degraded Titin fragment in serum reflects changes in muscle turnover induced by atrophy.

In this study we sought to determine whether a Titin peptide fragment can serve as a clinical biomarker for changes in muscle mass.

Species-specific cell-matrix interactions are essential for differentiation of alveoli like structures and milk gene expression in primary mammary cells of the Cape fur seal (Arctocephalus pusillus pusillus)

Few models are in place for analysis of extreme lactation patterns such as that of the fur seals which are capable of extended down regulation of milk production in the absence of involution. During a 10&ndash;12 month lactation period, female fur seals suckle pups on shore for 2&ndash;3 days, and then undertake long foraging trips at sea for up to 28 days, resulting in the longest intersuckling bouts recorded. During this time the mammary gland down regulates milk production. We have induced Cape fur seal (<i>Arctocephalus pusillus pusillus</i>) mammary cells in vitro to form mammospheres up to 900 &mu;m in diameter, larger than any of their mammalian counterparts. Mammosphere lumens were shown to form via apoptosis and cells comprising the cellular boundary stained vimentin positive. The Cape fur seal GAPDH gene was cloned and used in RT-PCR as a normalization tool to examine comparative expression of milk protein genes (&alpha;S2-casein, &beta;-lactoglobulin and lysozyme C) which were prolactin responsive. Cape fur seal mammary cells were found to be unique; they did not require Matrigel for rapid mammosphere formation and instead deposited their own matrix within 2 days of culture. When grown on Matrigel, cells exhibited branching/stellate morphogenesis highlighting the species-specific nature of cell&ndash;matrix interactions during morphological differentiation. Matrix produced in vitro by cells did not support formation of human breast cancer cell line, PMC42 mammospheres. This novel model system will help define the molecular pathways controlling the regulation of milk protein expression and species specific requirements of the extracellular matrix in the cape fur seal.<br />

Anti-metastatic and differential effects on protein expression of epigallocatechin-3-gallate in HCCLM6 hepatocellular carcinoma cells

Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and the third highest cause of cancer-related mortality in humans. Epigallocatechin-3-gallate (EGCG) has been shown to inhibit the metastatic activity of certain cancer cells. The aim of this study was to determine the effects and molecular mechanism(s) of action of EGCG in human HCC cells. A migration and invasion assay for the metastatic behavior of HCCLM6 cells was performed. The anti-metastatic effects of EGCG were investigated by RT-PCR and gelatin zymography. A total cellular protein profile was obtained using 2-dimensional gel electrophoresis (2-DE), followed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) analyses of proteins with significant differences in expression following treatment with EGCG. The results revealed that EGCG induced apoptosis and inhibited the metastasis of HCCLM6 cells. The anti-metastatic effects of EGCG were associated with the inhibition of matrix metalloproteinase (MMP)-2 and MMP-9 activity. The expression levels of far upstream element (FUSE) binding protein 1 (FUBP1), heat shock protein beta 1 (HSPB1), heat shock 60 kDa protein 1 (chaperonin) (CH60) and nucleophosmin (NPM) proteins, which are associated with metastasis, were significantly altered in the EGCG-treated HCCLM6 cells. The data from the present study suggest that EGCG has potential as a therapeutic agent for the treatment of HCC.

Conditions to ensure the 2-D characteristic polynomial of uncertain matrix is very strict Hurwitz

This article derives some new conditions for the bivariate characteristic<br />polynomial of an uncertain matrix to be very strict Hurwitz. The uncertainties are assumed of the structured and unstructured type. Using the two-dimensional inverse Laplace transform, we derive the bounds on the uncertainties, which will ensure that the bivariate characteristic polynomial is very strict Hurwitz. Two numerical examples are given to illustrate the results.<br />

Effect of the manufacturing process on the interlaminar fracture toughness of 2/2 twill weave fabric carbon/epoxy composites

A 2/2 twill weave fabric carbon fibre reinforced epoxy matrix composite MTM56/CF0300 was used to investigate the effect of different manufacturing processes on the interlaminar fracture toughness. Double cantilever beam tests were performed on composites manufactured by hot press, autoclave and 'Quickstep' processes. The 'Quickstep' process was recently developed in Perth, Western Australia for the manufacture of advanced composite components. The values of the mode I critical strain energy release rate (G1d were compared and the results showed that the composite specimens manufactured by the autoclave and the 'Quickstep' process had much higher interlaminar fracture toughness than the specimen produced by the hot press. When compared to specimens manufactured by the hot press, the interlaminar fracture toughness values of the Quickstep and autoclave samples were 38% and 49% higher respectively. The 'Quickstep' process produced composite specimens that had comparable interlaminar fracture toughness to autoclave manufactured composites. Scanning electron microscopy (SEM) was employed to study the topography of the mode I interlaminar fracture surface and dynamic mechanical analysis (DMA) was performed to investigate the fibre/matrix interphase. SEM micrography and DMA spectra indicated that autoclave and 'Quickstep' produced composites with stronger fibre/matrix adhesion than hot press.<br />

An experimental study of low velocity impact response in 2/2 twill weave composite laminates manufactured by a novel fabrication process

A novel fabrication process for advanced composite components&mdash;the QuicktepTM process was described. 2/2 twill weave MTM56/CF0300 carbon epoxy composite laminates were manufactured by the Quickstep and the autoclave processes. The response of these laminates to drop-weight low velocity impact at energy levels ranging from 5 to 30 J was investigated. It was found that the laminates fabricated by the Quickstep had better impact damage tolerance than those fabricated by the autoclave. Optical microscopy revealed extensive matrix fracture in the center of the backside of the autoclave laminates indicating the more brittle property of the epoxy matrix cured by the autoclave process. Interfacial shear strength (IFSS) for two composite systems were measured by micro&ndash;debond experiments. The MTM56/CF0300 material cured by the Quickstep showed stronger fibre matrix adhesion. Since the thickness and density of the impact targets produced by two processes were different, finite element analysis (FEA) was performed to study the effect of these factors on the impact response. The simulation results showed that the difference in thickness and density affects the stress distribution under impact loading. Higher thickness and lower density caused by processing lead to less endurance to drop weight impact loading. Therefore the better performance of Quickstep laminates under impact loading was not due to the thickness and density change, but resulted from stronger mechanical properties.<br />

Equal channel angular pressing of metal matrix composites: effect on particle distribution and fracture toughness

An Al6061-20%Al₂O₃ powder metallurgy (PM) metal matrix composite (MMC) with a strongly clustered particle distribution is subjected to equal channel angular pressing (ECAP) at a temperature of 370 &deg;C. The evolution of the homogeneity of the particle distribution in the material during ECAP is investigated by the quadrat method. The model proposed by Tan and Zhang [Mater Sci Eng 1998;244:80] for estimating the critical particle size which is required for a homogeneous particle distribution in PM MMCs is extended to the case of a combination of extrusion and ECAP. The applicability of the model to predict a homogeneity of the particle distribution after extrusion and ECAP is discussed. It is shown that ECAP leads to an increase of the&nbsp; uniformity of the particle distribution and the fracture toughness.<br />

Manufacturing influence on the delamination fracture behavior of the T800H/3900-2 carbon fiber reinforced polymer composites

'Torayca' T800H/3900-2 is the first material qualified on Boeing Material Specification (BMS 8-276) which utilizes the thermoplastic-particulate interlayer toughening technology. Two manufacturing processes, the autoclave process and the fast heating rated Quickstep&trade; process, were employed to cure this material. The Quickstep process is a unique composite production technology which utilizes the fast heat transfer rate of fluid to heat and cure polymer composite components. The manufacturing influence on the mode I delamination fracture toughness of laminates was investigated by performing double cantilever beam tests. The composite specimens fabricated by two processes exhibited dissimilar delamination resistance curves (R-curves) under mode I loading. The initial value of fracture toughness GIC-INIT was 564 J/m2 for the autoclave specimens and 527 J/m2 for the Quickstep specimens. However, the average propagation fracture toughness GIC-PROP was 783 J/m2 for the Quickstep specimens, which was 2.6 times of that for the autoclave specimens. The mechanism of fracture occurred during delamination was studied under scanning electron microscope (SEM). Three types of fracture were observed: the interlayer fracture, the interface fracture, and the intralaminar fracture. These three types of fracture played different roles in affecting the delamination resistance curves during the crack growth. More fiber bridging was found in the process of delamination for the Quickstep specimens. Better fiber/matrix adhesion was found in the Quickstep specimens by conducting indentation-debond tests.<br />

Stability of 2-D characteristic polynomials

This paper derives some new conditions for the bivariate characteristic polynomial of an uncertain matrix to be very strict Hurwitz. The uncertainties are assumed of the structured and unstructured type. By using the two-dimensional (2-D) inverse Laplace transform, the bounds on the uncertainties are derived which will ensure that the bivariate characteristic polynomial to be very strict Hurwitz. Two numerical examples are given to illustrate the results.<br />