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Mitochondrial Function

Mitochondrial oxidative phosphorylation generates 20-fold greater amounts of adenosine triphosphate (ATP) through pyruvate oxidation to CO2 than anaerobic glycolysis, which produces ATP by enzymatic reactions from glucose to pyruvate. In the kidney, proximal tubules are a ...

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Products of Arachidonic Acid Metabolism

Segmentation of the nephron relative to transport mechanisms and secretory activity has been recognized for decades, beginning with the pioneering studies of Alfred Newton Richards (1). Nephron segmentation regarding transcellular sodium and water movement has been subject ...

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Detection of Cysteine S-Nitrosylation and Tyrosine 3-Nitration in Kidney Proteins

Nitric oxide (NO) is a diatomic free-radical product of mammalian cells that has diverse and important physiological functions, including the regulation of renal, cardiovascular, pulmonary, gastrointestinal, neuronal, inflammatory, and immune systems. At the cellular level, ...

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Methods for Measurements of Heme Oxygenase (HO) Isoforms-Mediated Synthesis of Carbon Monoxide and HO-1 and HO-2 Proteins

As the key enzyme in heme degradation, heme oxygenase activity governs cellular heme concentration. Heme oxygenase catalyzes the conversion of heme to carbon monoxide and bilirubin with the release of iron, which can drive the synthesis of ferritin for iron sequestration (1,2). This is the ma ...

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The Juxtamedullary Nephron Preparation

Major aspects of renal function such as glomerular filtration, blood flow autoregulation, tubular reabsorption, and medullary concentrating ability rely on an exquisite paracrine-autocrine control of renal microvascular resistances, and on the coordinated network beha ...

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Tubuloglomerular Feedback

As originally described by Golgi, one consistent anatomical feature of all mammalian and many vertebrate kidneys is that a segment of the distal tubule is firmly attached to the vascular pole of its own glomerulum (1). This attachment is formed early in development, and is maintained throughout ...

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Microdissected Perfused Vessels

In vitro microperfusion was developed in order to study the transport of solutes and water across nephrons (1,2). The approach was later extended to renal microvessels for the purpose of measuring vasomotion of glomerular arterioles (3). The purpose of this chapter is to provide a general desc ...

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Renal Microperfusion Techniques

The advent of the technique of the isolated perfused tubule allowed renal physiologists to examine the transport characteristics of all portions of the nephron for the first time. The understanding of renal function prior to the advent of in vitro microperfusion rested on the integrative u ...

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How to Design a Clinical Trial

Clinical research, like all research, stems from curiosity. Although some research questions are harder to answer than others, the application of a well-designed trial coupled with the right question can yield valuable information. Indeed, a poorly designed randomized trial will not g ...

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Mapping the Disease

Bacon’s idealistic description of a research process, as naive and utopian as it sounds today, actually depicts the many functions of a single investigator, from literature search and collection of data to attempts of summing-up the investigation: the merchant, the miner, the depredator and ...

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The Measurement of GLUT4 Translocation in 3T3-L1 Adipocytes

Type 2 diabetes (T2D) is one of the fastest growing threats to human health in westernised and developing countries and is associated with central obesity, atherosclerosis, dyslipidaemia, hyperinsulinaemia and hypertension. Insulin resistance, defined as a diminished response ...

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Pancreatic Remodeling: Beta-Cell Apoptosis, Proliferation and Neogenesis, and the Measurement of Beta-Cell Mass and of Individual Beta-Cell Size

The endocrine pancreas has a significant remodeling capacity that plays a crucial role in the maintenance of glucose homeostasis. Changes in beta-cell apoptosis, replication and size, and islet neogenesis contribute to the remodeling of the endocrine pancreas. The extent of their res ...

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Morphology of Pancreatic Islets: A Time Course of Pre-diabetes in Zucker Fatty Rats

The Zucker fatty rat (fa/fa; ZR) is considered as a model for pre-diabetes, as characterised by a genetic defect in the leptin receptor, which results in hyperphagia, insulin resistance, hyperinsulinaemia, hyperlipoproteinaemia, and obesity. These animals become glucose intolera ...

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Fluorescent Immunohistochemistry and In Situ Hybridization Analysis of Pancreas

To facilitate the immunological reaction of antibodies with antigens in fixed tissues, it is necessary to unmask or retrieve the antigens through pretreatment of the specimens. However, adjustment of heat-induced antigen retrieval is always required for different tissues and anti ...

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The Measurement of Insulin Secretion Using Pancreas Perfusion in the Rodent

Under in vivo conditions, the study of physiological and pharmacological functions of an organ is difficult due to whole-body interactions with the organ. Thus, an in vitro technique for the perfusion of isolated pancreata was developed for physiologic and response studies including the ...

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HyperinsulinemicEuglycemic Clamp to Assess Insulin Sensitivity In Vivo

Insulin resistance, the impaired ability of insulin to stimulate glucose utilization, is a major characteristic of type 2 diabetes. Insulin sensitivity can be measured using a variety of techniques that are commonly employed in diabetes research and care. Of these, hyperinsulinemic- ...

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10 Lipoprotein(a) Quantitation

Lipoprotein(a) (Lp) was first discovered in 1963, by Kare Berg, as a variant of the beta-lipoproteins. Many years later, Lp(a) has been identified as an independent risk factor for the development of coronary heart disease (1). During the last two decades the structure and composition of Lp(a) has been i ...

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Isolation and Characterization of Glycosaminoglycans from Human Atheromatous Vessels

The complex cascade of events leading to the formation of atheromatous plaques depends on the interaction between several cell types, growth factors, cytokines, and molecules of the extracellular matrix (ECM) (1). Among different molecules of the ECM involved in atherogenesis, the gly ...

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Culture of Human Smooth Muscle Cells

The wall of a human artery consists of three distinct tunics. The tunica intima is lined by a layer of endothelial cells facing the lumen. Smooth muscle cells (SMCs) are the predominant cell type in the tunica media of arteries. They are surrounded by a basal lamina containing collagen IV, proteoglyca ...

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Culture of Human Endothelial Cells

Endothelial cells line the luminal surface of all blood vessels in the body. The endothelial surface in adult humans is composed of approximately l-6�l013 cells and covers an area of 1-7 m2. Endothelium serves many functions, including fluid and solute exchange through cell contraction, pro ...

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