RESUMEN
We characterized a full length L1 mRNA in a rheumatoid arthritis (RA) synovial tissue and determined the degree of methylation of its 5'-UTR. We asked whether not only intact but also altered L1s can exert biological activities by transfecting RA synovial fibroblasts (SF) with either retrotransposition-competent or incompetent L1s and examined their capacity to induce p38delta. Total RNA was isolated from the synovial tissue of a 35-year-old woman with highly destructive RA. A complete L1 sequence was obtained by 3'/5'-RACE. Methylation of the genomic 5'-UTR was determined by the sodium-disulfide/PCR method. RA-SF were transfected by lipofection with either a functional L1 or an ORF2-mutated L1 element. The expression of p38delta was measured by RT-PCR and Western blot. The full length L1 mRNA included a 5'-UTR, an ORF1 and an ORF2. Three of five CpG islands (60%) of the genomic L1 5'-UTR were hypomethylated and the ORF2 was deactivated by the insertion of stop codons. Both, intact and ORF2-mutated L1 vectors, induced the expression of p38delta. Thus, even an ORF2-mutated L1 element, as expressed in RA, is biologically active and both L1 ORF1 and p38delta transcripts may appear as a consequence of genomic hypomethylation. The induction of p38delta appears to be mediated by an ORF1/p40-dependent process. This is the first indication of a p40 mediated transactivation.
Asunto(s)
Proteínas de Unión al ADN/metabolismo , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Retroelementos/genética , Artritis Reumatoide/genética , Artritis Reumatoide/metabolismo , Western Blotting , Proteínas de Unión al ADN/genética , Humanos , Inmunohistoquímica , Hibridación in Situ , Proteína Quinasa 13 Activada por Mitógenos , Proteínas Quinasas Activadas por Mitógenos/genética , Filogenia , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
We compared the resistance to delamination and to adhesive/abrasive wear of conventional and highly cross-linked polyethylene tibial inserts of a cruciate-retaining total knee design using a knee simulator. Both groups were tested after aggressive, accelerated aging, and 1 set of conventional inserts was studied without aging. Aging oxidized the conventional, but not the highly cross-linked, inserts. The simulated normal gait testing lasted for 5 and 10 million cycles for the conventional and highly cross-linked inserts, respectively. Aged conventional inserts showed delaminations, whereas none were observed in the unaged conventional and aged cross-linked inserts. Wear rates measured by the gravimetric method were 9 +/- 2 mm3, 10 +/- 4 mm3, and 1 +/- 0 mm3 per million cycles; by the metrologic method, they were 8 +/- 1 mm3, 9 +/- 2 mm3, and 3 +/- 0 mm3 for the unaged conventional, aged conventional, and aged highly crosslinked inserts, respectively. In the test model used, oxidation led to delamination, whereas increased cross-link density resulted in reduced adhesive/abrasive wear of tibial inserts.