Although there is a great deal of interest in women's health, research on the health and well being of women with disabilities has not increased. In this article we present internal and structural barriers to wellness activities experienced by women with disabilities. We also discuss women's actual and recommended strategies to address these barriers. Data were collected in six focus groups in urban and rural Ontario, Canada. The participants represented a diversity of disability, age, and ethnoracial backgrounds. Our findings suggest that individual and structural barriers exist for the women, with structural barriers (physical, informational, and systemic access) being predominant. Barriers prevented women from engaging in desired wellness activities. Women discussed actual strategies to address these barriers, such as collective efforts to buy nutritious foods and recommendations to create greater access (e.g., increase health professionals' training in disability issues).

The hormone leptin is implicated in the regulation of appetite and body weight in rodents, primates and humans. We reported that the leptin gene (ob) is expressed in the brain, but the factors which control ob expression in the central nervous system are not known. We previously showed that brain-derived rat C6 glioblastoma cells express ob mRNA and protein. In the present study we examined the regulation of ob expression in C6 cells. Leptin and leptin receptor immunoreactivity was detected in C6 cells, suggesting a possible autocrine role for leptin. The identity of the leptin immunoreactivity (OB-ir) in C6 cells was confirmed by immunoprecipitation and Western blotting using two leptin specific polyclonal antibodies. Using RT-PCR analysis a product of the expected size for the short, but not the long, leptin receptor isoform was detected in C6 cells. Cells were maintained in serum-free (SF) media for 0–24 h in the presence of various regulators of leptin expression. Leptin mRNA levels were significantly higher in cells treated with dbcAMP (1 mM), IGF 1 (100 ng/ml) or insulin (5 μg/ml) compared to SF controls. In contrast, corticosterone (10−7M) reduced leptin mRNA. In the presence of dbcAMP, C6 cells undergo a dramatic alteration in morphology which is coincident with an apparent increase in the number of leptin-ir nuclei and an increase in leptin immunoreactivity. In contrast to C6 cells, glucocorticoids are reported to increase leptin levels in adipocytes/adipose tissue, while increases in intracellular cAMP levels are reported to reduce leptin levels. Overall, our in vitro data suggest that the regulation of leptin gene expression in C6 glioblastoma cells is different from that in adipocytes.

460 Direct antigen presentation by porcine lymphocytes and endothelial cells to human T cells requires the appropriate interaction of porcine costimulatory molecules with their human counterreceptors on the T cell. We have isolated the gene for the porcine costimulatory molecule CD40 in order to further characterize its interaction with human T cells. We designed degenerate CD40 primers based on homology between the published mouse and human sequences. RNA was isolated from porcine splenocytes stimulated with TPA and forskolin, reverse transcribed and amplified by polymerase chain reaction(PCR) using the degenerate primers. The 5' and 3' ends of the porcine CD40 gene were isolated using rapid amplification of cDNA ends (RACE) and were sequenced using standard dideoxy sequencing. Porcine CD40 has an open reading frame of 836 base pairs corresponding to a protein of 279 amino acids. Comparison of the pig and human CD40 sequences revealed 82% homology at the amino acid level. A search of all deposited genbank sequences revealed that porcine CD40 was closest in homology to the human CD40. Semiquantitative RTPCR was used to analyze the level of CD40 mRNA in different tissues. CD40 mRNA was detected in TPA stimulated splenocytes and faintly in unstimulated splenocytes. Porcine aortic endothelial cells (PAEC) passaged twice expressed significant levels of CD40 mRNA. We have isolated the porcine CD40 gene. The porcine gene is highly homologous to human CD40 gene and is expressed in endothelial cells. This will allow us to study the interactions between swine CD40 and human CD40L in direct costimulation of T cells and endothelial activation.

A family of 16 genes encoding the mouse ribosomal protein S24 was identified, and four members from this family were cloned. A single expressed intron-containing S24 gene (termed mrpS24) and one pseudogene (mrpS24p) were completely sequenced and characterized. The mrpS24 gene has seven exons and six introns spanning over 5.1 x 10(3) nucleotides (nt). The cap site of S24 was mapped to a G residue four nt upstream of a polypyrimidine tract and 15 nt downstream of a TATA-like (TATGA) element. The 5' region (-325 to +33) of the mrpS24 gene has a functional promoter that was able to express the fused chloramphenicol acetyltransferase (CAT) reporter gene. Two different forms of mouse S24 cDNA clones were previously isolated. Sequence analysis showed that one of these cDNA clones (termed S24a) lacks the entire exon V sequence (18 nt), and the deduced amino acid sequence is missing a C-terminal lysine residue encoded by the other cDNA (S24b). The pseudogene mrpS24p is flanked by an 11-bp direct repeat, and its sequence is almost identical to the S24 cDNA sequence, but it lacks two mini-exons, V and VI (20 nt), as in the cases of the human and rat S24 cDNAs. RT-PCR experiments demonstrated the existence of a third form (S24c) that similarly lacks both of the mini-exons, and suggested that different species of S24 mRNA might arise from alternative splicing of the mini-exons V and VI. Northern blot analysis showed that S24 expression is down- and up-regulated during adipocyte differentiation and in cellular transformation, respectively. RNase protection assays and RT-PCR experiments suggested that these cell-specific changes of S24 mRNA levels are mainly due to fluctuations in S24c mRNA level. Our results provide the first indication that a ribosomal protein gene is regulated by alternative usage of two mini-exons in a cell-specific manner.