DOI
https://doi.org/10.25772/3B3T-6A15
Defense Date
1996
Document Type
Thesis
Degree Name
Master of Science
Department
Biochemistry
First Advisor
Richard Brandt
Abstract
Determining the fragility of the red blood cell (RBC) is important for the diagnosis of and evaluation for treatment of several RBC diseases. In part RBC production is controlled through the hormone erythropoietin secreted by the kidney. In a previous study from this laboratory, it was found that RBC were more stable in uremic male rats compared to controls. In this experiment, uremia was induced in four groups of female rats through a two stage nephrectomy. The nephrectomy involved the removal of two-thirds of the left kidney, followed by the removal of the entire right kidney one week later. The animals were divided into three groups; NX-(5/6 nephrectomy), SH-(sham surgery), and PF-(sham surgery, but were fed the same food weight as the NX animals). The samples obtained in Trial I and Trial II were divided into two categories; initial and final. The initial samples were collected 14 days after the sham and five- sixth nephrectomy surgeries. The final sample were collected at the time of sacrifice. The samples obtained in Trial 11 consisted only of initial samples, taken fourteen days after the five-sixth nephrectomy and sham surgeries were completed. The samples in Trial IV were final samples, obtained at the time of sacrifice. Decreasing hypotonic %NaCl solutions were used to determine the hemolysis of RBC from the rats. RBC hemolysis was determined spectrophotometrically by monitoring hemoglobin absorbance at 540nm in the supernatant fluid. Analytic precision experiments using multiple assays of the same blood sample for 50% RBC hemolysis showed a coefficient of variation of only 1.1%. Analysis of the %NaCl at 50% RBC hemolysis did not differ significantly between the three groups of animals suggesting that although the NX animals were uremic, the RBC did not differ in stability to hypoosmotic shock. Future direction for this type of research will be extended to human studies where kidney failure patients (dialysis patients) can have both the age of the RBC and their fragility determined under therapy.
The erythrocyte hemolysis peroxide test (HPT) was also performed on rat blood samples from Trial IV and on five human blood samples, in order to determine hemolysis in the RBC. A 2% H2O2 solution was used to determine RBC stability and %Hemolysis was calculated by dividing the value for hemolysis due to H2O2 by the 100% hemolysis value and multiplying by 100. Analysis of the %Hemolysis by HPT for each animal sample also showed no significant difference between the three subgroups of animals. Future direction for this type of research will be extended to include all four trials of animals as well as human studies involving patients with kidney failure (dialysis patients).
Rights
© The Author
Is Part Of
VCU University Archives
Is Part Of
VCU Theses and Dissertations
Date of Submission
11-29-2017
Comments
Scanned, with permission from the author, from the original print version, which resides in University Archives.