Comparison of single-step and two-step radiofrequency rewarming of vitrified rabbit kidney and human kidney specimens

Cryobiology ◽  
2020 ◽  
Vol 97 ◽  
pp. 270
Author(s):  
Prem K. Solanki ◽  
Yoed Rabin
1960 ◽  
Vol 199 (5) ◽  
pp. 788-792 ◽  
Author(s):  
Carlos A. Schaffenburg ◽  
Erwin Haas ◽  
Harry Goldblatt

In a study of the renin-angiotensinogen interaction, under various conditions of time, temperature and concentration of renin and angiotensinogen, an indirect method for the quantitative determination of renin in the kidneys and of angiotensinogen in the serum of nine species was developed. This method involves the bioassay of the angiotensin formed as a result of the incubation in vitro of purified renal extracts with homologous serum or plasma. Renin and angiotensinogen, at the concentrations of the latter existing in the serum of the various species, were both found to be rate-determining in the process of angiotensin formation. There were striking differences in the concentration of renin in normal kidneys of the various species, with rabbit kidney having more than 70 times as much renin as human kidney. A pronounced increase of renin was found in the ischemic kidney of dogs with experimental renal hypertension. An exceptionally high concentration of angiotensinogen was present in normal human serum compared to that of the eight other species investigated. The results of the previous and present determinations of the renin concentration in the kidneys of various species differ considerably because the former method depended upon a heterologous renin-angiotensinogen reaction, in vivo, while the present evaluation is based on a homologous renin-angiotensinogen reaction in vitro.


1973 ◽  
Vol 136 (3) ◽  
pp. 659-668 ◽  
Author(s):  
Anne M. S. Grant ◽  
Albert Neuberger

1. The turnover rate of urinary Tamm–Horsfall glycoprotein in rabbits was determined by two different methods. The first involved measurement of the pool size of the glycoprotein in rabbit kidney and the daily urinary excretion rate by a radioimmunoassay from which the turnover rate was calculated. 2. The second method made use of the incorporation in vivo of Na214CO3 and sodium [14C]acetate. After a single intramuscular injection of one of these compounds, urine collections were made every 24h and the glycoprotein was isolated and its specific radioactivity was determined. 3. Incorporation of the label into urinary HCO3−, urea and plasma fibrinogen was also examined. The specific radio-activities of the O-acetyl, sialic acid, aspartic acid and glutamic acid residues isolated from the Tamm–Horsfall glycoprotein were compared and their half-lives were compared with that of the intact glycoprotein. The two methods gave results in quite close agreement and indicated a half-life for the glycoprotein of approx. 9h. 4. An attempt was made to localize the glycoprotein within the kidney and within the cell. It is present throughout the kidney, but was not detected in the brush-border fraction isolated from the proximal tubules. From differential cell-centrifugation studies, the glycoprotein seemed to be predominantly present in the soluble fraction (100000g supernatant). This suggests that it is either largely a soluble cytoplasmic component or is very loosely bound to a membrane, being readily released under the gentlest homogenization procedure. 5. The half-life of Tamm–Horsfall glycoprotein in human kidney was found by the radioimmunoassay method to be approx. 16h. The similarity between the composition of Tamm–Horsfall glycoprotein and human erythropoietin is discussed.


2017 ◽  
Vol 140 (1) ◽  
Author(s):  
Lili E. Ehrlich ◽  
Gregory M. Fahy ◽  
Brian G. Wowk ◽  
Jonathan A. Malen ◽  
Yoed Rabin

This study focuses on thermal analysis of the problem of scaling up from the vitrification of rabbit kidneys to the vitrification of human kidneys, where vitrification is the preservation of biological material in the glassy state. The basis for this study is a successful cryopreservation protocol for a rabbit kidney model, based on using a proprietary vitrification solution known as M22. Using the finite element analysis (FEA) commercial code ANSYS, heat transfer simulations suggest that indeed the rabbit kidney unquestionably cools rapidly enough to be vitrified based on known intrarenal concentrations of M22. Scaling up 21-fold, computer simulations suggest less favorable conditions for human kidney vitrification. In this case, cooling rates below −100 °C are sometimes slower than 1 °C/min, a rate that provides a clear-cut margin of safety at all temperatures based on the stability of rabbit kidneys in past studies. Nevertheless, it is concluded in this study that vitrifying human kidneys is possible without significant ice damage, assuming that human kidneys can be perfused with M22 as effectively as rabbit kidneys. The thermal analysis suggests that cooling rates can be further increased by a careful design of the cryogenic protocol and by tailoring the container to the shape of the kidney, in contrast to the present cylindrical container. This study demonstrates the critical need for the thermal analysis of experimental cryopreservation and highlights the unmet need for measuring the thermophysical properties of cryoprotective solutions under conditions relevant to realistic thermal histories.


2001 ◽  
Vol 12 (6) ◽  
pp. 1107-1113
Author(s):  
SATINDER S. SARANG ◽  
MATTHEW D. PLOTKIN ◽  
STEVEN R. GULLANS ◽  
BRIAN S. CUMMINGS ◽  
DAVID F. GRANT ◽  
...  

Abstract. The properties and functions of γ-aminobutyric acid (GABAA) receptors in the mammalian central nervous system are well studied. However, the presence and significance of GABAA receptors in nonneural tissue is less clear. The goal of this study was to examine the expression and localization of the GABAA receptor β2 and β3 subunits in the kidney. Reverse transcriptase products from RNA isolated from rat and rabbit kidney cortex and cerebellum and rabbit S2 segments were amplified by use of PCR and GABAA β2 and β3 subunit—specific primers. Sequencing of the kidney PCR products revealed that the rat kidney cortex and rat neuronal GABAA receptor β2 subunit were identical in nucleotide composition. The rabbit kidney and rabbit neuronal GABAA receptor β2 subunit were 99% identical in nucleotide composition. Sequencing of the kidney PCR products revealed that the rat kidney cortex and rat neuronal GABAA receptor β3 subunits were 93% and 95% identical in nucleotide and amino acid composition, and rabbit kidney cortex and rabbit neuronal GABAA receptor β3 subunits were 95% and 98% identical in nucleotide and amino acid composition, respectively. PCR screening of a human kidney cDNA library and sequencing revealed that the human kidney cortex and neuronal β3 subunits were identical in nucleotide composition. Immunoblot analysis of rat kidney cortex and brain identified immunoreactive proteins in the 55 to 57 kD region, corresponding to the GABAA receptor β2 and β3 subunits. Immunohistochemistry revealed cytosolic and basolateral staining of the proximal convoluted and straight tubule. These results provide compelling evidence for the expression of the GABAA receptor β2 and β3 subunits in the kidney of multiple species and the localization of the β2/β3 subunits to the renal proximal tubule.


Author(s):  
A. LeFurgey ◽  
P. Ingram ◽  
L.J. Mandel

For quantitative determination of subcellular Ca distribution by electron probe x-ray microanalysis, decreasing (and/or eliminating) the K content of the cell maximizes the ability to accurately separate the overlapping K Kß and Ca Kα peaks in the x-ray spectra. For example, rubidium has been effectively substituted for potassium in smooth muscle cells, thus giving an improvement in calcium measurements. Ouabain, a cardiac glycoside widely used in experimental and clinical applications, inhibits Na-K ATPase at the cell membrane and thus alters the cytoplasmic ion (Na,K) content of target cells. In epithelial cells primarily involved in active transport, such as the proximal tubule of the rabbit kidney, ouabain rapidly (t1/2= 2 mins) causes a decrease2 in intracellular K, but does not change intracellular total or free Ca for up to 30 mins. In the present study we have taken advantage of this effect of ouabain to determine the mitochondrial and cytoplasmic Ca content in freeze-dried cryosections of kidney proximal tubule by electron probe x-ray microanalysis.


2005 ◽  
Vol 173 (4S) ◽  
pp. 300-300
Author(s):  
Sreedhar Sagi ◽  
Lutz Trojan ◽  
Peter Aiken ◽  
Maurice S. Michel ◽  
Thomas Knoll

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