Assays for Measurement of MMPs and TIMPs in Body Fluids

Although the first MMP was described in remodeled amphibian tissue in 1962 (G12), it was not until 1986 that MMPs were identified in circulating blood. MMPs were initially recognized in plasma inadvertently, as a contaminant during the chemical isolation of fibronectin (J1). MMP-2 and MMP-9 were subsequently identified as normal components of plasma using gelatin zymography, which depicted these elements as negative staining bands of gelatinolytic activity in gelatin-impregnated SDS...

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No correlation was found between pathological TNM staging histological grade and plasma MMP-9 levels in patients with renal cancer (L3). Tumor tissue levels of MMPs were consistent with the concept that plasma MMP-9 was primarily released from the kidney. In contrast, plasma MMP-2 and TIMP-2 concentrations were actually higher in healthy controls than in renal cancer patients. Zucker et al. reported results of plasma MMP-7 measurements using an ELISA. A bimodal...

General Considerations and Future Directions

There is increasing evidence to suggest that the presence of micrometas-tases has prognostic relevance. However, the impact of micrometastasis on the prognosis varies depending on the body compartment (blood, lymph nodes, and peritoneal lavage fluids), tumor origin, and methods used to detect micrometastasis. As for the body compartment and tumor origin, the evidence to date seems more convincing for peritoneal lavage fluids in gastric cancer than for peripheral blood and lymph nodes in...

Monitoring the Papai System

The following section provides a brief description of analytical systems typically used in the qualitative and quantitative measurement of PAs and PAIs. The advantages and limitations of each technique are discussed. It should be noted, however, that the usefulness of any of these techniques is dependent on the specific application in the biological system of interest. 2.1. Enzyme-Linked Immunosorbent Assay Although enzyme-linked immunosorbent assays (ELISAs) are commercially available and...

Biomarkers for Increased Risk of Developing Estrogen Initiated Cancer

Many types of markers are being evaluated to estimate risk of developing breast cancer. These include breast density body mass index expression of BRCA1 and or BRCA2 gene cytological changes in breast epithelial cells collected by ductal lavage and levels of estrogens and androgens in serum, breast tissue, or nipple aspirate fluid. Some studies suggest that levels of selected estrogen metabolites, estrogen conjugates, and or depurinating estrogen-DNA adducts in breast ductal fluid, collected...

Oxidation Products of Lipids and Proteins and Measurement Methods

Lipid Peroxidation Cannabinoid

Oxidation of polyunsaturated fatty acids (PUFA) in lipoproteins may be mediated by reactive species such as radicals, transition metals, other elec-trophiles, and by enzymes. Once initiated, oxidation of lipids may proceed by a chain reaction, illustrated in Fig. 4 (R5). In step 1, an oxidant captures an electron from a PUFA to produce a lipid radical. In step 2, after rearrangement, the conjugated diene radical reacts rapidly with singlet oxygen to produce a lipid peroxide radical, which is...

Conclusions

MMPs and TIMPs are involved in numerous physiologic and pathologic processes. In spite of considerable understanding of the chemistry and biology of MMPs, their role in disease processes is incompletely understood. Increased levels of MMPs and TIMPs have been demonstrated in the blood and urine of patients with many different types of disease. A compendium of descriptive reports has documented increased plasma levels of MMP-9 in patients with heart disease, stroke, cancer, skin disease,...

MMPs Identified in Urine of Patients with Cancer

Gelatin zymography has been used to demonstrate the presence of several forms of MMPs in the urine of patients with cancer. The potential for using this test for early diagnosis or staging of cancer has been proposed. Based on the molecular weight of MMP-2 and MMP-9 (72 and 92 kDa, respectively) and the glomerular filtration limit of 45 kDa, it was not anticipated that MMPs would be detected in urine. Nonetheless, employing gelatin substrate zymography and immunoassays, Marguilies et al. (M6)...

Detailed Procedures for Some Methods that Measure Oxidation Products in Plasma

Although none of the methods described here are simply mix-and-read, they are methods that are straightforward enough to be performed in a clinical laboratory using equipment that is generally available. A difficulty encountered in the past was ex vivo oxidation. This problem has been eliminated by adding BHT to specimens and reagents, including those for OxLDL and MDA-LDL. Except for studying susceptibility to oxidation, we collect specimens in EDTA purple type tubes and bring plasma or serum...

Prophylaxis

The term prophylaxis as used in this chapter is limited to medical counter-measures applied relatively shortly before penetration of a toxic agent into the organism. There is a question what will happen after the administration of the prophylactic drug. When the treatment is unnecessary, it can be described as prophylaxis. However, though successful prophylaxis can be observed for some OP, full protection of the organism without post-exposure treatment, especially for soman poisoning, remains...

Quantitative Detection of Micrometastases and Its Prognostic Significance

Many reports have been published on the quantitative detection of micro-metastasis as determined by real-time RT-PCR in the peripheral blood, bone marrow, lymph nodes, and peritoneal washes, although the reports on its prognostic relevance are few. Overviews of currently available results are summarized in Table 1 to Table 3. The evidence to date seems to be more convincing for peritoneal lavage fluids than for peripheral blood and lymph nodes in gastrointestinal malignancies. Thus, this...

MRNA Extraction from Living Cells

Most cells can change their patterns of gene expression in response to extracellular stimuli. For example, fibroblast cells at G0 state change their gene expression pattern in response to various growth factors for cell proliferation. Relative mRNA levels of more than 500 genes change in response to stimulation I3 . The expression pattern of each gene shows different profiles. For example, the mRNAs for immediate early genes are induced within 15 min after stimulation and are terminated in 20...

Modification of AFM Tips

Epoxy Resin Synthesis

Our method using an AFM can be applied to extracting other biomole-cules as well as mRNA from living cells. When we try to identify specific biomolecules on the cell surface, we can use modified AFM tips that interact with our interest. We introduced here the method for AFM tip modification Fig. 9 . 5.1. Modification of AFM Tips with Amino Groups 1. The silicon or silicon nitride tips were cleaned with ethanol for 5 min, followed by rinsing with Milli-Q water for 5 min. 2. They were treated...