Springer protocols feed by Molecular Medicine 
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This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.The Isolation and Quantitation of Fetuin-A-Containing Calciprotein Particles from Biological Fluids
Multiple overlapping systemic and local inhibitory networks have evolved to prevent the unwanted deposition of mineral at ectopic sites. Fetuin-A is a liver-derived glycoprotein abundant in plasma that binds and stabilizes nascent mineral ion nuclei to form soluble colloidal high molecular weight complexes, called calciprotein particles (CPP). The binding of fetuin-A to mineral retards crystal ripening and precipitation from the aqueous phase, thereby facilitating the regulated clearance of mineral debris from the extracellular fluid. However, persistent disturbances in this humoral homeostatic system, as frequently seen i...
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
An In Vitro Murine Model of Vascular Smooth Muscle Cell Mineralization
Vascular calcification (VC) is seen ubiquitously in aging blood vessels and prematurely in disease states like renal failure. It is thought to be driven by a number of systemic and local factors that lead to extra-osseous deposition of mineral in the vascular wall and valves as a common endpoint. The response of resident vascular smooth muscle cell to these dystrophic signals appears to be important in this process. Whilst in vivo models allow the observation of global changes in a pro-calcific environment, identifying the specific cells and mechanisms involved has been largely garnered from in vitro experiments, which pro...
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Vascular Calcification in Uremia: New-Age Concepts about an Old-Age Problem
A hallmark of aging, and major contributor to the increased prevalence of cardiovascular disease in patients with chronic kidney disease (CKD), is the progressive structural and functional deterioration of the arteries and concomitant accrual of mineral. Vascular calcification (VC) was long viewed as a degenerative age-related pathology that resulted from the passive deposition of mineral in the extracellular matrix; however, since the discovery of “bone-related” protein expression in calcified atherosclerotic plaques over 20 years ago, a plethora of studies have evoked the now widely accepted view that VC is a...
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Prolonged and Continuous Measurement of Kidney Oxygenation in Conscious Rats
A relative deficiency in kidney oxygenation, i.e., renal hypoxia, may contribute to the initiation and progression of acute and chronic kidney disease. A critical barrier to investigate this is the lack of methods allowing measurement of the partial pressure of oxygen in kidney tissue for long periods in vivo. We have developed, validated, and tested a novel telemetric method that can do this. Here we provide details on the calibration, implantation, implementation for data recording, and reuse of this telemetry-based technology for measurement of medullary tissue oxygen tension in conscious, unrestrained rats. This techni...
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Intravital Multiphoton Imaging of the Kidney: Tubular Structure and Metabolism
Multiphoton microscopy (MPM) allows the visualization of dynamic pathophysiological events in real time in live animals. Intravital imaging can be applied to investigate novel mechanisms and treatments of different forms of kidney disease as well as improve our understanding of normal kidney physiology. Using rodent models, in conjunction with endogenous fluorescence and infused exogenous fluorescent dyes, measurement can be made of renal processes such as glomerular permeability, juxtaglomerular apparatus function, interactions of the tubulointerstitium, tubulovascular interactions, vascular flow rate, and the renin-angio...
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Assessment of Renal Hemodynamics and Oxygenation by Simultaneous Magnetic Resonance Imaging (MRI) and Quantitative Invasive Physiological Measurements
In vivo assessment of renal perfusion and oxygenation under (patho)physiological conditions by means of noninvasive diagnostic imaging is conceptually appealing. Blood oxygen level-dependent (BOLD) magnetic resonance imaging (MRI) and quantitative parametric mapping of the magnetic resonance (MR) relaxation times T
2* and T
2 are thought to provide surrogates of renal tissue oxygenation. The validity and efficacy of this technique for quantitative characterization of local tissue oxygenation and its changes under different functional conditions have not been systematically examined yet and remain to be established. For thi...
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Magnetic Resonance Imaging (MRI) Analysis of Ischemia/Reperfusion in Experimental Acute Renal Injury
Imbalance between renal oxygen delivery and demand in the first hours after reperfusion is suggested to be decisive in the pathophysiological chain of events leading to ischemia-induced acute kidney injury. Here we describe blood oxygenation level-dependent (BOLD) magnetic resonance imaging (MRI) for continuous monitoring of the deoxyhemoglobin-sensitive MR parameter T
2* in the renal cortex, outer medulla, and inner medulla of rats throughout renal ischemia/reperfusion (I/R). Changes during I/R are benchmarked against the effects of variations in the fraction of inspired oxygen (hypoxia, hyperoxia). This method may be use...
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Biopsychronology: A Method Using Live Tissue Staining to Image Cell Function in the Kidney
Methods to monitor the status of a graft prior to transplantation are highly desirable to avoid unnecessary surgical interventions and follow-up treatments and to optimize the clinical outcome as delayed graft function may lead to costly and lengthy follow-up treatments or even organ loss. As a promising step in this direction we present a method which combines the use of fine needle biopsies, the staining of living cells with dyes suitable to monitor mitochondrial status/cellular integrity, and live confocal real-time analysis. (Source: Springer protocols feed by Molecular Medicine)
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Use of Cationized Ferritin Nanoparticles to Measure Renal Glomerular Microstructure with MRI
Magnetic resonance imaging (MRI) is becoming important for whole-kidney assessment of glomerular morphology, both in vivo and ex vivo. MRI-based renal morphological measurements can be made in intact organs and allow direct measurements of every perfused glomerulus. Cationic ferritin (CF) is used as a superparamagnetic contrast agent for MRI. CF binds to the glomerular basement membrane after intravenous injection, allowing direct, whole-kidney measurements of glomerular number, volume, and volume distribution. Here we describe the production, testing, and use of CF as an MRI contrast agent for quantitative glomerular morp...
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Decellularization of Rat Kidneys to Produce Extracellular Matrix Scaffolds
The extracellular matrix (ECM) retains three-dimensional structures for the stimulation of cell growth, with components of the ECM relatively conserved between species. Interest in the use of decellularized scaffold-based strategies for organ regeneration is increasing rapidly. Decellularized scaffolds derived from animal organs are a promising material for organ engineering, with a number of prominent advances having been reported in the past few years. (Source: Springer protocols feed by Molecular Medicine)
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Renal Sympathetic Denervation in Rats
Experimental renal sympathetic denervation is a well-established technique. Classically, renal sympathetic denervation is achieved by dorsal rhizotomy. While more recently, direct renal sympathetic denervation is typically applied by stripping all visible renal nerve bundles followed by painting with a solution of 10 % phenol in ethanol to remove the remaining nerves. In clinical settings, a reliable marker of renal sympathetic denervation or renal sympathetic overactivity has not been established. However, in experimental models, successful renal sympathetic denervation is validated by a decrease in renal norepinephrine c...
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Rat Models of Acute and/or Chronic Peritoneal Injuries Including Peritoneal Fibrosis and Peritoneal Dialysis Complications
We describe herein two rat models of peritoneal injury that we have recently proposed. (Source: Springer protocols feed by Molecular Medicine)
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Isolation and Propagation of Rat Peritoneal Mesothelial Cells
With the development of peritoneal dialysis in many countries, there has been much interest in the cell biology of peritoneal mesothelial cells. In this chapter we describe a reliable and reproducible method for the culture of rat primary mesothelial cells (RPMCs). This chapter outlines how to isolate mesothelial cells from rat peritoneum. The subculture of primary peritoneal mesothelial cells and the characterization by immunofluorescence is also described in detail. (Source: Springer protocols feed by Molecular Medicine)
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Propagation and Culture of Human Renal Fibroblasts
The renal fibroblast and phenotypically related myofibroblast are universally present in all forms of progressive kidney disease. The in vitro study of the fibroblast, its behavior, and factors affecting its activity is therefore key to understanding both its role and significance. In this protocol, we describe a reproducible method for selective propagation and culture of primary human renal fibroblasts from the human kidney cortex. Techniques for their isolation, subculture, characterization, and cryogenic storage and retrieval are described in detail. (Source: Springer protocols feed by Molecular Medicine)
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
Isolation and Primary Culture of Murine Podocytes with Proven Origin
Genetic studies on hereditary kidney diseases and in vivo experimental model studies have revealed a critical role for the podocyte in glomerular function and disease. Primary podocyte cultures as well as immortalized podocyte cell lines have been used extensively to study podocyte function. Although, primary cells often more closely resemble the in vivo cells, they may have only a finite replicative life span before they reach senescence. Therefore, the success of studies using primary cell cultures depends on standardized isolation and culture protocols that allow reproducible generation of stable primary cultures. (Sour...
Source: Springer protocols feed by Molecular Medicine - December 22, 2015 Category: Molecular Biology Source Type: news
