http://repub.eur.nl/res/aut/15311/ List of Publications en http://repub.eur.nl/static-eur/img/logo.png http://repub.eur.nl http://repub.eur.nl/res/pub/17057/ 2009-06-01T00:00:00Z A 43-yr-old male presented with a 6-month history of episodes of coughing, shortness of breath and fever. He suffered from dyspnoea on minor exertion. The patient worked in a cattle feed factory and noticed that he had more complaints after his working hours. His symptoms could be ascribed to hypersensitivity pneumonitis due to contact with phytase, an enzyme added to cattle feed to strengthen bone and diminish phosphorus excretion. The diagnosis was supported by bibasal lung crackles on physical examination, restrictive ventilatory defect (with decreased diffusion capacity for carbon monoxide), typical radiographical findings, lymphocytosis in bronchoalveolar lavage fluid and a positive exposure test performed at the workplace. Blood examination showed high immunoglobulin G levels to phytase. After treatment and cessation of phytase contact the patient became symptom free and lung function normalised. Phytase should be considered as a cause of occupational hypersensitivity pneumonitis in the animal feed industry. http://repub.eur.nl/res/pub/13437/ 2004-10-15T00:00:00Z Phosphorylation of the human AR (androgen receptor) is directly correlated with the appearance of at least three AR isoforms on an SDS/polyacrylamide gel. However, it is still not clear to what extent phosphorylation is involved in the occurrence of isoforms, which sites are phosphorylated and what are the functions of these phosphosites. The human AR was expressed in COS-1 cells and AR phosphorylation was studied further by mutational analyses and by using reversed-phase HPLC and MS. The reversed-phase HPLC elution pattern of the three isoforms revealed that Ser-650 was phosphorylated constitutively. After de novo synthesis, only Ser-650 was phosphorylated in the smallest isoform of 110 kDa and both Ser-650 and Ser-94 were phosphorylated in the second isoform of 112 kDa. The hormone-induced 114 kDa isoform shows an overall increase in phosphorylation of all the isolated peptides. The activities of the Ser-Ala substitution mutant S650A (Ser-650-->Ala) was found to be identical with wild-type AR activation in four different cell lines and three different functional analyses, e.g. transactivation, N- and C-terminal-domain interaction and co-activation by transcriptional intermediary factor 2. This was also found for mutants S94A and S515A with respect to transactivation. However, the S515A mutation, which should eliminate phosphorylation of the potential mitogen-activated protein kinase site, Ser-515, resulted in an unphosphorylated form of the peptide containing Ser-650. This suggests that Ser-515 can modulate phosphorylation at another site. The present study shows that the AR isoform pattern from AR de novo synthesis is directly linked to differential phosphorylation of a distinct set of sites. After mutagenesis of these sites, no major change in functional activity of the AR was observed.