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Overexpression of SGTP and/or MT may contribute to various carcinogenic processes and to resistance to anticancer treatment. The importance of these proteins, although clearly established in solid tumours, has not been fully understood in haematopoietic neoplasm. The aim of this study was to determine the expression of MT and SGTP in the bone marrow of patients with MPD. Twenty paraffin-embedded bone marrow core biopsy specimens from newly diagnosed patients with MPD were evaluated — osteomyelofibrosis (OMF), n = 9 and chronic myelocytic leukaemia (CML), n = 11. We demonstrate increased SGTP and MT expression in the bone marrow of MPD patients. In our study levels of MT in OMF patients were higher than in CML. This suggests that MT expression may correlate with bone marrow fibrosis. These data, although based on a relatively small number of patients, raise the possibility that SGTP and MT may play a role in the pathogenesis of MPD. The clinical significance of this phenomenon needs further investigation.
Ubc9 is a homologue of the E2 ubiquitin conjugating enzyme and participates in the covalent linking of SUMO-1 molecule to the target protein. In this report we describe a simple and efficient method for obtaining pure human recombinant Ubc9 protein. The purified Ubc9 retained its native structure and was fully active in an in vitro sumoylation assay with the promyelocytic leukaemia (PML) peptide as a substrate. In order to better understand the physiology of Ubc9 protein we examined its levels in several rat tissues. Immunoblot analyses performed on tissue extracts revealed quantitative and qualitative differences in the expression pattern of Ubc9. The Ubc9 protein was present at a high level in spleen and lung. Moderate level of Ubc9 was detected in kidney and liver. Low amount of Ubc9 was observed in brain, whereas the 18 kDaband of Ubc9 was barely visible or absent in heart and skeletal muscle. In heart and muscle extracts the Ubc9 antibodies recognized a 38 kDa protein band. This band was not visible in extracts of other rat tissues. A comparison of the relative levels of Ubc9 mRNA and protein indicated that the overall expression level of Ubc9 was the highest in spleen and lung. In spleen, lung, kidney, brain, liver and heart there was a good correlation between the 18 kDa protein and Ubc9 mRNA levels. In skeletal muscle the Ubc9 mRNA level was unproportionally high comparing to the level of the 18 kDa protein. The presented data indicate that in the rat the expression of the Ubc9 protein ap­pears to have some degree of tissue specificity.
Changes in blood leucocyte levels were investigated in Spraque-Dowley rats vaccinated with cDNA or protein of glutathione S-transferase (GST) of F. hepatica and subsequently challenged with metacercariae of the liver fluke. The analysis of the leucocyte responses measured in vaccinated rats suggests that the form of antigen used for vaccination intluenced dynamics of white blood cell response to the fluke infection. The most clear differences were observed in neutrophil and eosinophil levels. The weakest reaction of these cells to the challenge infection was observed in rats vaccinated twice with cDNA. In contrast, in rats which received the first antigen dose as cDNA and the second vaccination with GST protein, both neutrophil and eosinophil responses were much higher, especially at 5 and 9 WAI.
The role of glutathione-S-transferase (GST, EC 2.5.1.18) in biochemical host defence in experimental trichinellosis was evaluated. The activity of GST in mouse skeletal muscles was measured during the muscular phase of trichinellosis, starting from the 3rd week post infection (w.p.i.) to the 11th w.p.i. Activity was determined spectrophotometrically by monitoring the formation of thioether (S-2,4-dinitrophenylglutathione) from the reduced form of glutathione and 1-chloro-2,4-dinitrobenzene used as a substrate and as an example of xenobiotics. The changes in the activity of GST were as follows: an increase in activity starts in the 4th w.p.i., peaks (up to 310% of the normal value) in the 6th w.p.i., decreases in the 8th week and a final, weak rise was observed in week 11. The statistically significant changes in GST activity in this phase of experimental trichinellosis suggest that this enzyme participates in the biochemical defence of the host against Trichinella infection.
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