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Study of protein changes in patients with nephrotic syndrome and Anderson-Fabry disease
Vojtová, Lucie ; Zima, Tomáš (advisor) ; Teplan, Vladimír (referee) ; Průša, Richard (referee)
Background: Heavy proteinuria may be caused by either increased glomerulal basement membrane permeability or membrane or podocyte structural damage, and also by impairment of secretion-reabsorption tubular processes. In this study, 60 patients with nephrotic proteinuria and other diagnoses (lupus nephritis, membranous nephropathy, IgA nephropathy, Wegener's granulomatosis) and 20 patients with Anderson-Fabry disease (AFD),which is an X-linked genetic disorder with deficient a-galactosidase A activity, were analysed by the 2D electrophoresis method. The main aim of this work was to investigate possible differences in urine proteins in nephropaties, between healthy controls and AFD patients and to identify abnormal proteins as potential biomarkers of disease. Methods: The urine proteins were devided by isoelectric focusing method using polyacrylamide strips (pH 3-10 linear). The second dimensional SDS electrophoresis was performed in 12 % polyacrylamide gel. The proteins were visualized by silver method and selected proteins were identified by MALDI-TOF MS. The gels were evaluated by Phoretix 2D expression software 2005. Results: We found out that without adding protease inhibitors we can detect proteolysis, with increased quantity of proteins manifested in the area about 10 kDa and decreased quantity...
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Study of protein changes in patients with nephrotic syndrome and Anderson-Fabry disease
Vojtová, Lucie ; Zima, Tomáš (advisor) ; Teplan, Vladimír (referee) ; Průša, Richard (referee)
Background: Heavy proteinuria may be caused by either increased glomerulal basement membrane permeability or membrane or podocyte structural damage, and also by impairment of secretion-reabsorption tubular processes. In this study, 60 patients with nephrotic proteinuria and other diagnoses (lupus nephritis, membranous nephropathy, IgA nephropathy, Wegener's granulomatosis) and 20 patients with Anderson-Fabry disease (AFD),which is an X-linked genetic disorder with deficient a-galactosidase A activity, were analysed by the 2D electrophoresis method. The main aim of this work was to investigate possible differences in urine proteins in nephropaties, between healthy controls and AFD patients and to identify abnormal proteins as potential biomarkers of disease. Methods: The urine proteins were devided by isoelectric focusing method using polyacrylamide strips (pH 3-10 linear). The second dimensional SDS electrophoresis was performed in 12 % polyacrylamide gel. The proteins were visualized by silver method and selected proteins were identified by MALDI-TOF MS. The gels were evaluated by Phoretix 2D expression software 2005. Results: We found out that without adding protease inhibitors we can detect proteolysis, with increased quantity of proteins manifested in the area about 10 kDa and decreased quantity...
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Development, growth and variability of semiaquatic bug Microvelia reticulata (Heteroptera: Gerromorpha: Veliidae)
VOJTOVÁ, Lucie
I investigated the effect of environmental factors (temperature and food availability) on the development and growth of semiaquatic bug Microvelia reticulata Burmeister, 1835. I focused on the developmental rate isomorphy. In a cross-designed experiment, M. reticulata was individually kept over whole juvenile period (eggs to reaching maturity) and subjected to treatments of three temperature (17, 21, 25 °C) and two food availability (full - daily feeding and reduced - 2 days interval) and their interaction. Development and growth was strongly influenced by temperature. Longest postembryonic development measured in individuals kept in the lowest temperature (54 days) contrasted to those kept in the highest temperature (16 days). Mortality was also significantly influenced by temperature; highest mortality rate was found in the 17 °C treatment. Effect of food availability was significant only in low temperature, resulting in the better survival of individuals on the high food level. In other temperature treatments, I found no effect of food on survival rate of M. reticulata. However, the food level influenced development significantly. In the 21 °C and 25 °C treatment, individuals on a full food developed faster than individuals on a reduced food. In 17 °C the food availability had no effect on time required to reach maturity. The best fitness (expressed as combination of low mortality rate and short development) was observed in 21 °C, both other temperature treatment were found stressing. Food availability had stronger effect on female individuals compared to males. In conclusion, I found no evidence of developmental rate isomorphy in M. reticulata. The research was supported by project of the Grant Agency of the Czech Republic GAČR P505/10/0096.
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