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Iron regulatory protein 1 (IRP1) post-transcriptionally regulates the expression of proteins involved in the iron metabolism of mammals. IRP1 is a bifunctional cytosolic protein which can exhibit aconitase activity or bind to iron responsive element (IREs) in the untranslated regions of specific mRNAs. The modulation of IRP1 activities and its consequence for intracellular iron homeostasis is best characterized in rodents and humans. Little is known about IRP1 in farm animals. In this study, we analyzed the two activities of IRP1 in the livers of four farm animal species (cattle, goat, pig and rabbit) and their relationship to hepatic iron content. We found an inverse correlation between spontaneous IRP1 IRE binding activity and non-haem iron content in the liver. Using the electrophoretic mobility shift assay, we showed differential mobility of IRE/IRP1 complexes formed with hepatic cytosolic extracts from various farm animal species. We discuss this observation in relation to a comparative analysis of mammalian IRP1 amino acid sequences.
Piglets were euthanized on 1st, 4th, 7th, 14th, and 28th day of life. Iron dextran was injected intramuscularly (200 mg Fe/head) on the 4th day. The concentration of Fe, Ca, Mg, Mn, Zn, and Cu was estimated by flame atomic absorption spectrometry. The liver Fe concentration was 177 ± 49 mg Fe/kg on the 1st day, and this may be considered to be adequate. After injecting iron dextran, it increased nine-fold to 1594 ± 11 mg Fe/kg fresh liver, which was higher than the toxic level reviewed in literature for adult pigs (400 mg Fe/kg). The liver iron decreased to the first day level only on day 28. It was significantly correlated (r = 0.81) with the liver Mn concentration. Calcium in liver (25.0 ± 6.3 mg/kg) increased on day 7 (55.0 ± 2.7 mg/kg) and remained on the same level until the end of the experiment (55 mg/kg). No significant changes were observed in liver Mg content (about 200 mg/kg in average). Apart from the iron content, the most evident changes were stated in the liver Zn level which rapidly decreased from 99.0 ± 8.9 (1st day) to 19.2 -21.8 mg/kg (7-28 days). The liver Cu content remained unchanged for two weeks after birth (64.9-52.6 mg/kg), and only on day 28 had decreased three times as compared to the first day (21.2 ± 1.7 mg/kg). Generally, the liver Fe concentration increased to the toxic level after intramuscular injection of iron dextran (200 mg/head). Hence, it would be better for piglet health to apply iron dextran in two doses. It seems that there is a close correlation in the metabolism of iron and manganese in the liver. The liver concentration of other elements was not affected by iron injections. The Cu deposit in the liver is sufficient only for the first two weeks of life, which may be another risk factor of anaemia in piglets.
The purpose of the study was to elaborate a molecular procedure to identify the DUMPS mutation which causes hereditary disorder (deficiency of uridine monophosphate synthase) in cattle. The degree of this disorder was determined in a gene pool of a Polish bredcattle population which consisted of 1038 animals: 781 artifical insemination bulls from all centers in Poland and 257 sire dams from the two largest breeding centers of Holstein-Friesian cattle: Osięciny and Osowa Sień. The diagnosis was performed using the PCR-RFLP method and technique based on automated solid-phase sequencing of PCR products. In the examined material no DUMPS carrier was found. This indicates that the population of cattle reared in Poland is free from this economically significant disorder. The results obtained indicate a high effectiveness of monitoring of point mutation DUMPS by the PCR-RFLP method. Moreover, it is worth noting the wide possibilities of examining other biologically significant genome regions using the solid-phase sequencing method.
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