996-11 Direct Gene Transfer and Expression with Arterial Iontophoretic Catheter Delivery

Iontophoresis is a technique of molecular delivery which uses electric current to enhance movement of charged molecules into tissues. A porous balloon catheter was tested with a central silver chloride electrode capable of generating a potential gradient across the arterial wall using an adhesive patch placed on the skin to serve as the anode. We hypothesized that this catheter delivery system might effectively transfer negatively charged plasmid DNA into arterial cells in vivo.MethodsTo localize plasmid DNA arterial delivery, a 7 Fr iontophoretic porous balloon catheterwast,’aced into porcine carotid arteries underflouroscopic guidance. 15μg of 35S-Iabeled plasmid DNA (1.4×106 cpm/μg) expressing the heat stable human alkaline phosphatase (hAP) gene with an RSV promoter was infused through the balloon at 6 atm pressure. A constant current density of 2.5 mA/cm2 was maintained for 10 minutes. The ;35S-labeled plasmid DNA delivery was repeated on the contralateral carotid artery under identical conditions with the absence of electric current. 20 minutes after gene transfer, the arteries were fixed in situ and processed for autoradiography. To analyze gene transfer and expression, 8 porcine carotid arterial segments were subject to iontophoretic gene delivery for 10 minutes at 6 atm with a current density of 2.5 mA/cm2 using the RSV hAP plasmid (n=6) or control plasmid (n=2). Animals were sacrificed 5 days after gene delivery and the transfected arteries analyzed by PCR and heat stable alkaline phosphatase histochemistry.ResultsAutoradiography of the arteries which underwent ;35S-labeled plasmid delivery revealed minimal radiolabel in the luminal cells of the control artery in which current was not delivered. In contrast, significant amounts of radiolabel were present in the media and adventitia of the artery subject to current delivery. PCR analysis of the arterial segments studied 5 days after delivery confirmed gene transfer in all hAP segments and was negative in control arteries. Staining for heat stable recombinant alkaline phosphatase activity demonstrated recombinant protein expression in 5% of medial cells and 10% of adventitial cells in arteries which underwent hAP gene transfer. Control arteries were negative for hAP staining.ConclusionsIontophoretic catheter gone delivery can be used to perform direct plasmid DNA delivery with expression of recombinant protein in medial and adventitial cells

Longitudinal direct medical costs associated with constipation in women: Constipation direct costs in women

Although direct medical costs for constipation related medical visits are thought to be high, to date there have been no studies examining longitudinal resource utilization in adults with constipation

Vascular dysfunction in aged mice contributes to persistent lung fibrosis

Idiopathic pulmonary fibrosis (IPF) is a progressive disease thought to result from impaired lung repair following injury and is strongly associated with aging. While vascular alterations have been associated with IPF previously, the contribution of lung vasculature during injury resolution and fibrosis is not well understood. To compare the role of endothelial cells (ECs) in resolving and non‐resolving models of lung fibrosis, we applied bleomycin intratracheally to young and aged mice. We found that injury in aged mice elicited capillary rarefaction, while injury in young mice resulted in increased capillary density. ECs from the lungs of injured aged mice relative to young mice demonstrated elevated pro‐fibrotic and reduced vascular homeostasis gene expression. Among the latter, Nos3 (encoding the enzyme endothelial nitric oxide synthase, eNOS) was transiently upregulated in lung ECs from young but not aged mice following injury. Young mice deficient in eNOS recapitulated the non‐resolving lung fibrosis observed in aged animals following injury, suggesting that eNOS directly participates in lung fibrosis resolution. Activation of the NO receptor soluble guanylate cyclase in human lung fibroblasts reduced TGFβ‐induced pro‐fibrotic gene and protein expression. Additionally, loss of eNOS in human lung ECs reduced the suppression of TGFβ‐induced lung fibroblast activation in 2D and 3D co‐cultures. Altogether, our results demonstrate that persistent lung fibrosis in aged mice is accompanied by capillary rarefaction, loss of EC identity, and impaired eNOS expression. Targeting vascular function may thus be critical to promote lung repair and fibrosis resolution in aging and IPF.Bleomycin‐induced lung injury promotes transient fibrosis accompanied by increased capillary density in young mice. In contrast, persistent fibrosis, capillary rarefaction, loss of endothelial cell identity, and reduction of Nos3 are observed in aged mice. eNOS/NO signal is an important driver of fibroblast quiescence and fibrosis resolution, that is lost with aging. Lung vascular bed plays a critical role during lung repair and fibrosis resolution.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/156458/2/acel13196_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/156458/1/acel13196.pd

Primary care incidence and treatment of four neuropathic pain conditions: A descriptive study, 2002–2005

<p>Abstract</p> <p>Background</p> <p>Between 1992 and 2001 the UK general practice incidence of post-herpetic neuralgia and trigeminal neuralgia declined, whilst the incidence of painful diabetic neuropathy increased. The most common first line treatments were compound analgesics. As therapeutic options have subsequently changed, this study presents updated data on incidence and prescribing patterns in neuropathic pain.</p> <p>Methods</p> <p>A descriptive analysis of the epidemiology and prescription treatment at diagnosis of incident post-herpetic neuralgia (n = 1,923); trigeminal neuralgia (1,862); phantom limb pain (57) and painful diabetic neuropathy (1,444) using computerised UK general practice records (THIN): May 2002 to July 2005.</p> <p>Results</p> <p>Primary care incidences per 100,000 person years observation of 28 (95% confidence interval (CI) 27–30) for post-herpetic neuralgia, 27 (95%CI 26–29) for trigeminal neuralgia, 0.8 (95%CI 0.6–1.1) for phantom limb pain and 21 (95%CI 20–22) for painful diabetic neuropathy are reported. The most common initial treatments were tricyclic antidepressants (post-herpetic neuralgia) or antiepileptics (trigeminal neuralgia and painful diabetic neuropathy) and opioid analgesics (phantom limb pain). The mean number of changes before a stable drug regimen was 1.2 to 1.5 for trigeminal neuralgia, painful diabetic neuropathy and post-herpetic neuralgia, and 2.4 for phantom limb pain.</p> <p>Conclusion</p> <p>The incidence of phantom limb pain and post-herpetic neuralgia are decreasing whilst painful diabetic neuropathy plateaued and trigeminal neuralgia remained constant. Despite more frequent use of antidepressants and antiepileptics for first line treatment, as opposed to conventional non-opioid analgesics, changes to therapy are common before a stable regimen is reached.</p