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Studies of gene expression by different islet endocrine cell populations can provide useful information about signal transduction cascades regulating α-, β- and δ-cell function. Experiments on expression of β-cell gene products are relatively easy to perform in rodent islets as these ...

In vitro culture has been well defined as a useful tool to improve survival and functionality of isolated islets of Langerhans. Evaluation of islet morphology and function is essential prior to use for experimental investigations. Novel techniques such as co-culture and the use of matrices ...

Human β-cell gene profiling is a powerful tool for understanding β-cell biology in normal and pathological conditions. The assessment is complicated when isolated islets are studied because of contamination by non-β-cells and the exposure to the trauma of isolation that causes changes ...

Transdifferentiation is defined as an irreversible switch in postnatal life of one differentiated cell to another. Transdifferentiation from different cellular origins into pancreatic-like β-cells is of clinical significance since this approach may offer a potential cure f ...

As a research tool, medical imaging has considerably increased our understanding of joint inflammation in vivo. Although conventional radiography remains important for diagnosis, other imaging modalities such as magnetic resonance imaging (MRI) and ultrasonography show co ...

The development in the techniques for obtaining synovial tissue biopsy, especially through arthroscopy, have resulted in greater access to high-quality synovial tissue. The use of immunohistochemistry in arthritis research has greatly furthered our understanding of the vari ...

Arthroscopy continues to experience a growth in interest from the rheumatology community reflecting a common desire to gain better understanding of the underlying processes in inflammatory and degenerative joint diseases. Arthroscopy provides the ability to assess the inter ...

The chapter focuses on the detection of specific mRNA by in situ hybridization (ISH) in synovial tissue specimens. This technique is widely applied, reliable, specific, and sensitive, because even small quantities of mRNA can be detected. Presented here contemporary protocols for ISH us ...

Most current approaches used to analyze gene expression in tissue samples are based on RNA isolated either from cultured synovial cells or from synovial biopsies. However, this strategy does not distinguish between specific gene expression profiles of cells originating from discre ...

Many studies in arthritis research require an evaluation of the cellular responses within the joint and the ensuing matrix degradation in articular cartilage. The early histochemical/histological scale of Mankin (1) has been widely used but recently challenged as insufficient (2). ...

Quantitative image analysis is a form of imaging that includes microscopic histological quantification, video microscopy, image analysis, and image processing. Hallmarks are the generation of reliable, reproducible, and efficient measurements via strict calibration and s ...

In rheumatoid arthritis (RA), the synovium represents the predominant site of inflammation and joint destruction and is regarded as the key organ involved in disease pathogenesis. It has been studied in different ways over the last 30 yr, yielding information about the mechanisms involved ...

Necrosis and apoptosis have been demonstrated in articular cartilage in response to trauma and disease. However, cell death in articular cartilage may also be thought of as a scale of cell death culminating in secondary necrosis with the failure to remove apoptotic cells from the tissue. The in s ...

Assay of collagenase activity involves the use of radiolabeled collagen. Stimulation of cartilage with proinflammatory cytokines results in the upregulation of collagenases and the subsequent release of degraded collagen fragments. These enzymes can be localized in both oste ...

Quantitation of glycosaminoglycans (GAGs) in the form of aggrecan fragments released from cartilage in culture is a simple way to determine the efficacy of different cytokines alone or in combination in simulating cartilage catabolism. Two approaches for GAG assay are described, with ...

Two methods for the assessment of the expression of gelatinases A and B, MMP-2 and MMP-9, in articular cartilage are described. Immunohistochemical analysis of tissue sections provides information about the precise localization of the enzymes within the tissue, pinpointing the cells t ...

Membrane-type 1 matrix metalloproteinase (MT1-MMP) is a type I transmembrane protein which exhibits biological activities on the cell surface. One of the characteristic functions of this enzyme is activation of proMMP-2 on the cell surface (1). This process can be monitored using gelatin ...

This chapter provides practical information on the assay of tissue inhibitor of metalloproteinase (TIMP) activity and background information enabling meaningful interpretation of the data. Protocols are given for assessing the presence of TIMPs in biological samples by immun ...

Osteoclasts are bone-resorbing multinucleated cells derived from the monocytemacrophage lineage. The authors have developed a mouse marrow culture system and a coculture system of mouse osteoblasts and hemopoietic cells, in which osteoclasts are formed in response to various o ...

Rodent arthritis models are tools to study joint destruction. This chapter describes methods to prepare and analyze histological sections from inflamed paws. Protocols to create decalcified paraffin-embedded as well as undecalcified plastic-embedded sections are provide ...