Immunoendocrine responses over a three week international rugby union series.

AIM: This study evaluated changes in immunoendocrine makers over an international series in professional rugby union players (N.=8). METHODS: Venous bloods were taken on camp-entry, as well as before and after (0, 14 and 38 h) two games spaced over 21-days. Samples were analysed for changes in serum C-reactive protein (CRP), cortisol (C), testosterone (T), blood leukocytes, interleukin 6 (IL-6) and creatine kinase (CK). RESULTS: Significant reductions in CK activity and CRP concentrations were evident on day 5 (pre-game 1) when compared to camp-entry (day 1); P<0.05. A large acute-phase response was observed following both games. Differences in the magnitude of this response appeared dependant on the number of collisions players experienced during play. Compared to camp-entry, sharp increases in C (40%) and decreases (37%) in T were evident after both games; P<0.05. A gradual increase in T/C ratio was observed throughout the tournament; values 35% and 45% higher on days 19 and 21 than those observed at camp-entry (P<0.05). CONCLUSION: Current data suggests that improved physiological recovery can be achieved during an international rugby union series. Monitoring of previous club activity is essential to ensure optimal player readiness prior to participation in international rugby union games.

Molecular analysis of functional and nonfunctional genes for human ferrochelatase: isolation and characterization of a FECH pseudogene and its sublocalization on chromosome 3.

A pseudogene related to the functional gene (FECH) for the heme biosynthetic enzyme ferrochelatase (ferroheme-protolyase; EC 4.99.1.1.) was isolated from a human genomic library using a ferrochelatase cDNA hybridization probe. The pseudogene shows > 80% overall nucleotide sequence identity to the functional gene (including the 3' untranslated region and polyadenylation signals) but contains no intronic sequences in the region corresponding to the open reading frame of expressed ferrochelatase. Furthermore, the pseudogene sequence contains small deletions and insertions creating frameshifts and numerous termination codons, indicating that it does not encode a functional polypeptide. Northern blot analysis using pseudogene-specific probes failed to demonstrate transcripts in samples of human erythroid cell RNA in which ferrochelatase mRNA was readily detected. Southern blot experiments using restriction endonuclease-digested human genomic DNA probed either with ferrochelatase-specific cDNA fragments or pseudogene-specific genomic sequences confirmed the presence of distinct loci for the expressed and nonfunctional genes, respectively. Localization of the human ferrochelatase pseudogene to 3p22-p23 was determined by fluorescent metaphase chromosomal hybridization in situ using three genomic clones in lambda EMBL3 spanning a contiguous region of approximately 30 kb. This newly identified locus, distinct from the expressed FECH gene, on 18q22, is characteristic of a processed human pseudogene. The existence of the ferrochelatase pseudogene has practical implications for the molecular analysis of mutations responsible for erythropoietic protoporphyria in man.

Molecular characterization of a ferrochelatase gene defect causing anomalous RNA splicing in erythropoietic protoporphyria.

Erythropoietic protoporphyria is an inherited disorder caused by deficient activity of the enzyme ferrochelatase. We have examined the ferrochelatase gene in an 11-year-old female with protoporphyria and have found that she is heterozygous for a mutation at a conserved residue in the exon 3 donor splice site consensus sequence (T(+2)-->G). This is inherited from her father, who also has deficient ferrochelatase activity. As a consequence of the mutation, ferrochelatase transcripts are aberrantly spliced and give rise to mRNA molecules in which sequences corresponding to exon 3 are absent. This leads to the expression of a ferrochelatase protein lacking a central region of 40 amino acids.

Assignment of the human ferrochelatase gene (FECH) and a locus for protoporphyria to chromosome 18q22.

We have mapped the human gene for ferrochelatase (FECH; ferroheme-protolyase, EC 4.99.1.1) to chromosome 18 by hybridization of cDNA to sorted chromosomes. The probe was obtained by PCR-directed amplification of a human marrow cDNA library in lambda gt 10. Subchromosomal localization of ferrochelatase to 18q22 was determined by chromosomal hybridization in situ using a human ferrochelatase genomic clone in lambda EMBL 3 that contained a 20-kb insert. Since ferrochelatase activity is deficient in patients with the inherited disease erythropoietic protoporphyria, a locus for this disease may be assigned to 18q22, one of few monogenic defects that have been mapped to this chromosome.