Hepatocyte gene therapy in a large animal: A neonatal bovine model of citrullinemia

The development of gene-replacement therapy for inborn errors of metabolism has been hindered by the limited number of suitable large-animal models of these diseases and by inadequate methods of assessing the efficacy of treatment. Such methods should provide sensitive detection of expression in vivo and should be unaffected by concurrent pharmacologic and dietary regimens. We present the results of studies in a neonatal bovine model of citrullinemia, an inborn error of urea-cycle metabolism characterized by deficiency of argininosuccinate synthetase and consequent life-threatening hyperammonemia. Measurements of the flux of nitrogen from orally administered 15NH4 to [15N]urea were used to determine urea-cycle activity in vivo. In control animals, these isotopic measurements proved to be unaffected by pharmacologic treatments. Systemic administration of a first-generation E1-deleted adenoviral vector expressing human argininosuccinate synthetase resulted in transduction of hepatocytes and partial correction of the enzyme defect. The isotopic method showed significant restoration of urea synthesis. Moreover, the calves showed clinical improvement and normalization of plasma glutamine levels after treatment. The results show the clinical efficacy of treating a large-animal model of an inborn error of hepatocyte metabolism in conjunction with a method for sensitively measuring correction in vivo. These studies will be applicable to human trials of the treatment of this disorder and other related urea-cycle disorders.

Potential roles of osteopontin and αVβ3 integrin in the development of coronary artery restenosis after angioplasty

Angioplasty procedures are increasingly used to reestablish blood flow in blocked atherosclerotic coronary arteries. A serious complication of these procedures is reocclusion (restenosis), which occurs in 30–50% of patients. Migration of coronary artery smooth muscle cells (CASMCs) to the site of injury caused by angioplasty and subsequent proliferation are suggested mechanisms of reocclusion. Using both cultured human CASMCs and coronary atherectomy tissues, we studied the roles of osteopontin (OPN) and one of its receptors, αvβ3 integrin, in the pathogenesis of coronary restenosis. We also measured the plasma levels of OPN before and after angioplasty and determined the effect of exogenous OPN on CASMC migration, extracellular matrix invasion, and proliferation. We found that cultured CASMCs during log phase of growth and smooth muscle cell layer of the coronary atherosclerotic tissues of patients express both OPN mRNA and protein at a significantly elevated level compared with controls. Interestingly, whereas the baseline plasma OPN levels in control samples were virtually undetectable, those in patient plasma were remarkably high. We also found that interaction of OPN with αvβ3 integrin, expressed on CASMCs, causes migration, extracellular matrix invasion, and proliferation. These effects were abolished when OPN or αvβ3 integrin gene expression in CASMCs was inhibited by specific antisense S-oligonucleotide treatment or OPN-αvβ3 interaction was blocked by treatment of CASMCs with antibodies against OPN or αvβ3 integrin. Our results demonstrate that OPN and αvβ3 integrin play critical roles in regulating cellular functions deemed essential for restenosis. In addition, these results raise the possibility that transient inhibition of OPN gene expression or blocking of OPN-αvβ3 interaction may provide a therapeutic approach to preventing restenosis.

CD4-independent, CCR5-dependent infection of brain capillary endothelial cells by a neurovirulent simian immunodeficiency virus strain

Brain capillary endothelial cells (BCECs) are targets of CD4-independent infection by HIV-1 and simian immunodeficiency virus (SIV) strains in vitro and in vivo. Infection of BCECs may provide a portal of entry for the virus into the central nervous system and could disrupt blood–brain barrier function, contributing to the development of AIDS dementia. We found that rhesus macaque BCECs express chemokine receptors involved in HIV and SIV entry including CCR5, CCR3, CXCR4, and STRL33, but not CCR2b, GPR1, or GPR15. Infection of BCECs by the neurovirulent strain SIV/17E-Fr was completely inhibited by aminooxypentane regulation upon activation, normal T cell expression and secretion in the presence or absence of ligands, but not by eotaxin or antibodies to CD4. We found that the envelope (env) proteins from SIV/17E-Fr and several additional SIV strains mediated cell–cell fusion and virus infection with CD4-negative, CCR5-positive cells. In contrast, fusion with cells expressing the coreceptors STRL33, GPR1, and GPR15 was CD4-dependent. These results show that CCR5 can serve as a primary receptor for SIV in BCECs and suggest a possible CD4-independent mechanism for blood–brain barrier disruption and viral entry into the central nervous system.

A influência do pensamento liberal de Benjamin Constant na formação do Estado Imperial Brasileiro / Cleber Francisco Alves. --

Inclui notas explicativas, bibliográficas e bibliografia.

Manager Training and Development Strategy: A Methodology to Link Strategic Initiatives at Washington and Lee University

Washington and Lee University has developed and implemented a strategic plan and performance development initiative for 2007-2017. In concert with the strategic plan and performance development initiative, supportive manager core competencies have been identified. The deliverable for this capstone project is a documented methodology that supports the strategy for the design and implementation of a manager training and development program that ensures needed competencies are available. The author uses survey data to determine training needs and priorities and, through a review of literature, investigates effective strategies to arrive at a successful implementation methodology. The author presents findings and conclusions regarding design implementation methodology for manager training and development.

Benjamin, BitTorrent, Bootlegs: auratic piracy cultures?

Seventy-five years ago, Walter Benjamin showed us that the line between "production" and "reproduction" had begun to blur. Reproduction was no longer optional, consequential and degrading (the shredding of the original’s aura), but was instead being transformed into a principle of production itself: something was produced bearing in mind how it was to be reproduced. No longer did the original exist (in photography, film, music recordings), but instead diffusion, exhibition. The work existed precisely at the time and place of its enjoyment. Today, the cultural pirates of the new digital era take this principle to the extreme, with a certain characteristic also foreseen by Benjamin: a yearning to participate, to post-produce something captured in order to later return it to the Internet, modified in some way and made available to others. This postproduction is what is now often mixed up with reception, just as production and reproduction used to be in Benjamin’s day. Postproduction on the receiver’s side, which somehow augments and extends the received work, in other words creates an etymologically rigorous author-ization (auctor as the root of both author and augmentation). The cultural pirate only deserves redemption thanks to this creative augmentation.

Accounts with Benjamin Huntington, 1785-1787

This folder contains two handwritten copies of the accounts between Reverend Andrew Croswell of Boston, and Croswell's executor William Croswell, and Benjamin Huntington, for money collected from Col. Gallup between 1785 and 1787.

Letter from Benjamin Croswell to William Croswell, 1810 March 30

One-page letter from William Croswell's cousin with information about the ailments of Benjamin Croswell's wife.

Letter from William Croswell to Benjamin Webb, 1820 January 3

Draft of a one-page letter with information on Croswell's activities.

Letter from William Croswell to Benjamin Webb, 1822 September 4

Draft of a one-page letter primarily concerning Croswell's Mercator maps.