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Transport of triglycerides (TGs) and cholesteryl esters (CEs) in plasma can be viewed as taking place via two major groups of lipoproteins, the TG-rich lipoproteins (chylomicrons and VLDL), on one hand, and the cholesterol-rich lipoproteins (LDL and HDL), on the other. The metabolism of these g ...

A number of epidemiological and clinical studies have linked elevated plasma homocysteine (Hcy) to atherosclerotic vascular disease affecting coronary, carotid, and peripheral vessels. Plasma Hcy can be considered a marker of methionine metabolic efficiency, mainly affect ...

Modified low density lipoprotein (LDL) is considered to be a risk factor for the development and progression of atherosclerosis (1-3). Glycated LDL and oxidized LDL are the only modified lipoproteins that exist naturally in the human body, but oxidized LDL does not exist in the circulation bec ...

Immortalization of chondrocytes increases life span and proliferative capacity but does not necessarily stabilize the differentiated phenotype. Expansion of chondrocyte cell lines in continuous monolayer culture may result in the loss of phenotype, particularly if high cell ...

Immortalized chondrocytes of human origin have been developed to serve as reproducible models for studying chondrocyte function. In this chapter, methods for immortalization of primary human chondrocytes with SV40-TAg, HPV-16 E6/E7, and telomerase by retrovirally mediated tr ...

It is well documented that adult cartilage has minimal self-repair ability. Current methods for treatment of cartilage injury focus on the relief of pain and inflammation and have met with limited long-term success. In the forefront of new therapeutic approaches, autologous chondrocy ...

Reverse transcriptase-polymerase chain reaction (RT-PCR) is a powerful, sensitive, and rapid method to monitor small amounts of nucleic acids. This is of particular interest for small amounts of cells, as in cartilage. We present here two protocols to isolate total RNA and a protocol to study ma ...

Unlike any other technology in molecular biology, the polymerase chain reaction (PCR) has changed the technological armamentarium of molecular scientists working on cartilage, in terms of outstanding sensitivity and accuracy. Four approaches to determine mRNA expression lev ...

The direct isolation of RNA from cartilage has often proved difficult owing to a number of factors. Cartilage has a low cell content and contains an extracellular matrix rich in proteoglycans, which copurify with the RNA as they are large and negatively charged macromolecules. In our laborato ...

In situ hybridization allows detection and localization of specific nucleic acid sequences directly within a cell or tissue. We present an in situ hybridization protocol using double-stranded DNA or single-stranded RNA probes labeled with to localize and visualize the temporal and s ...

Global gene expression profiling through the use of microarray technology is among the most powerful molecular biological techniques available to diabetes researchers today. In this chapter, we outline how to appropriately perform a microarray experiment using pancreatic isl ...

Microarray analysis has become a core part of biomedical research and its value can be seen in thousands of research papers. A successful microarray experiment needs to be augmented by specialized data mining techniques if the data are to be fully exploited. Here, tools that concentrate on three ...

This is not a “Methods” chapter in the traditional sense. Rather, it is an essay designed to help address one of the most frequently asked questions by investigators about to embark on a study requiring an animal model of diabetes - what is the “right” model for the reader’s specific research application. ...

In order to better understand the events which precede and precipitate the onset of type 2 diabetes (T2DM) several nutritional animal models have been developed. These models are generated by manipulating the diet of either the animal itself or its mother during her pregnancy and, in comparis ...

This chapter describes the detailed protocol for the isolation and purification of islets of Langerhans from rodent pancreas using collagenase digestion. The first step of the process is to separate and isolate the insulin-producing islets of Langerhans from the rest of the pancreas. The ...

This chapter describes a method to measure the viability of isolated intact islets of Langerhans from rat pancreas and considers the use of isolated islets and of pancreatic beta-cell lines to study cell viability following culture. The islet isolation method is based on the use of preparatio ...

Insulin secretion plays an essential part in the modulation of glucose homeostasis. Pancreatic β cells are extremely sensitive to small changes in the concentration of glucose, peptides, hormones and fatty acids and insulin secretion is stimulated in response to these factors. The meas ...

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 ...