Implementation of an Online Reporting System to Identify Unprofessional Behaviors and Mistreatment Directed at Trainees at an Academic Medical Center.

Importance: Unprofessional behaviors and mistreatment directed at trainees continue to challenge the learning environment. Academic medical institutions should encourage reports of inappropriate behavior and address such reports directly to create a safe learning environment.

Objective: To determine the feasibility of creating and implementing an online reporting system for receiving and reviewing complaints of unprofessional behavior directed toward or experienced by students, postdoctoral trainees, and residents.

Investigating a Quantitative Measure of Student Self-authorship for Undergraduate Medical Education.

Construct: In this study, the authors investigated the validity of a quantitative measure of self-authorship among medical students. Self-authorship is a cognitive-structural theory incorporating the ability to define one's beliefs, identity, and social relations to operate in a complex, ambiguous environment.

Background: Competency-based medical education (CBME) provides learners with the opportunity to self-direct their education at an appropriate pace to develop and exhibit required behaviors while incorporating functioning relationships with supervisors and trainers.

An update on the etiology of abdominal aortic aneurysms: implications for future diagnostic testing.

Abdominal aortic aneurysm (AAA) disease is multifactorial with both environmental and genetic risk factors. The current research in AAA revolves around genetic profiles and expression studies in both human and animal models. Variants in genes involved in extracellular matrix degradation, inflammation, the renin-angiotensin system, cell growth and proliferation and lipid metabolism have been associated with AAA using a variety of study designs.

An Evolving Understanding of the Genetic Causes of Abdominal Aortic Aneurysm Disease.

Aneurysms of the abdominal aorta (AAA) are relatively common - affecting as many as 8% of men and 1% of women over the age of 65. AAAs are characterized by a 50% increase in the diameter of the aneurysmal aorta compared with the normal vessel. Degeneration of structural components of the aortic wall is believed to be central in the pathogenesis of AAAs.

Use of a Pteridine Moiety to Track DNA Uptake in Cells.

Fluorescence labeled oligonucleotides have a long history of being used to monitor nucleic acid transport and uptake. However, it is not known if the fluorescent moiety itself physically limits the number of pathways that can be used by the cell due to steric, hydrophobic, or other chemical characteristics. Here, we report a method for comparing the uptake kinetics of oligonucleotides labeled either with the fluorescent pteridine, 3-methyl-8-(2-deoxy-β-D-ribofuranosyl) isoxanthopterin (3MI), or the common fluorophore 5-carboxyfluorescein (5-FAM).

Wicking: a rapid method for manually inserting ion channels into planar lipid bilayers.

The planar lipid bilayer technique has a distinguished history in electrophysiology but is arguably the most technically difficult and time-consuming method in the field. Behind this is a lack of experimental consistency between laboratories, the challenges associated with painting unilamellar bilayers, and the reconstitution of ion channels into them. While there has be a trend towards automation of this technique, there remain many instances where manual bilayer formation and subsequent membrane protein insertion is both required and advantageous.

Localization of the nucleic acid channel regulatory subunit, cytosolic malate dehydrogenase.

NACh is a nucleic acid-conducting channel found in apical membrane of rat kidney proximal tubules. It is a heteromultimeric complex consisting of at least two proteins: a 45-kDa pore-forming subunit and a 36-kDa regulatory subunit. The regulatory subunit confers ion selectivity and influences gating kinetics.

Presence of the nucleic acid channel in renal brush-border membranes: allosteric modulation by extracellular calcium.

We have previously described a cell surface channel complex that is highly selective for nucleic acid (6, 7). The channel complex was purified to homogeneity by solubilizing renal brush-border membranes (BBM) with CHAPS and separation by liquid chromatography. It was characterized by reconstitution in planar lipid bilayers.

Applications of gene therapy to kidney disease.

Purpose Of Review: This review summarizes recent applications of somatic cell gene therapy to the treatment of monogenetic renal diseases, renal cell carcinoma, and for the induction of tolerance in solid organ transplantation. In addition, several new gene therapy techniques will be discussed including gene and messenger RNA repair strategies, as well as methods designed to modify the expression of normal genes that may have application in the treatment of multigenetic disorders.

Recent Findings: Animal studies have demonstrated prolonged graft survival after the successful induction of tolerance to alloantigens via hematopoietic molecular chimerism.

Podocan, a novel small leucine-rich repeat protein expressed in the sclerotic glomerular lesion of experimental HIV-associated nephropathy.

Growing evidence suggests that human immunodeficiency virus (HIV)-1 infection of podocytes plays a central role in the glomerular disease of HIV-associated nephropathy (HIVAN). As an approach to identify host genes involved in the pathogenesis of the sclerotic glomerular lesion in HIVAN, representational difference analysis of cDNA was used to identify differentially expressed genes in HIV-1 transgenic and nontransgenic podocytes. We isolated a novel member of the small leucine-rich repeat (SLR) protein family, podocan, that is expressed at high levels in the HIV-1 transgenic podocytes.

Lentiviral gene transduction of kidney.

Gene transfer into kidney holds great potential as a novel therapeutic approach. We have studied the transduction of kidney in vivo after delivery of lentiviral vectors by various routes of administration. A lentiviral vector expressing the bacterial lacZ gene from the cytomegalovirus early promoter was used.

Cytosolic malate dehydrogenase confers selectivity of the nucleic acid-conducting channel.

We have described previously a cell surface channel that is highly selective for nucleic acids. Nucleic acid conductance is 10 pS and the channel is at least 10,000-fold more selective for oligodeoxynucleotides than any anion tested (1). Herein we provide evidence that the nucleic acid-conducting channel (NACh) is a heteromultimeric complex of at least two proteins; a 45-kDa pore-forming subunit (p45) and a 36-kDa regulatory subunit (p36).

Transduction of renal cells in vitro and in vivo by adeno-associated virus gene therapy vectors.

There has been an increasing interest recently in the possibility of treating renal diseases using gene therapy. The ability to pursue gene therapy for renal diseases has been limited by the availability of an adequate system for gene delivery to the kidney. Adeno-associated virus (AAV) is a defective virus of the parvovirus family that has a number of properties attractive for renal gene delivery: recombinant AAV contains no viral genes; expression of genes delivered by these vectors does not activate cell-mediated immunity; the virus is able to transduce nondividing as well as dividing cells; and both wild-type and recombinant AAV integrate into the host chromosome resulting in long-term gene expression.

Adeno-associated virus gene transfer into renal cells: potential for in vivo gene delivery.

The human parvovirus adeno-associated virus (AAV), type 2, has a number of features that make it an attractive choice as a vector for gene delivery to the kidney. AAV vectors permit long-term gene expression in vivo by integration into the host genome, have potential for site-specific integration on chromosome 19, do not express viral genes or generate a cellular immune response, and demonstrate enhancement of gene expression by chemotherapeutic agents that are approved for use in vivo. These properties confer advantages to AAV over other viral and nonviral methods for gene transfer.

Identification and characterization of a cell membrane nucleic acid channel.

We have identified a 45-kDa protein purified from rat renal brush border membrane that binds short single-stranded nucleic acid sequences. This activity was purified, reconstituted in proteoliposomes, and then fused with model planar lipid bilayers. In voltage-clamp experiments, the reconstituted 45-kDa protein functioned as a gated channel that allows the passage of nucleic acids.

Calcium regulation of a cell surface nucleic acid channel.

We have recently purified a 45 kDa protein from rat renal brush border membrane that functions as a macromolecular nucleic acid channel when reconstituted in artificial lipid bilayers. To explore the role of calcium in the regulation of this channel, purified protein was reconstituted in bilayers and calcium concentration was altered while voltage clamp experiments were performed. Open probability of the channel was less than 1% in 0 calcium and increased an average of 19-fold when calcium concentration was increased from 0 to 0.

Molecular therapy for renal diseases.

The introduction of molecular therapy through the delivery of nucleic acids either as oligonucleotides or genetic constructs holds enormous promise for the treatment of renal disease. Significant barriers remain, however, before successful organ-specific molecular therapy can be applied to the kidney. These include the development of methods to target the kidney selectively, the definition of vectors that transduce renal tissue, the identification of appropriate molecular targets, the development of constructs that are regulated and expressed for long periods of time, the demonstration of efficacy in vivo, and the demonstration of safety in humans.

Transport of phosphorothioate oligonucleotides in kidney: implications for molecular therapy.

The systemic administration of phosphorothioated antisense oligonucleotides has been demonstrated to be an effective strategy for the control of gene expression. Because previous studies have suggested both hepatic and renal accumulation of systemically administered oligonucleotides, we explored whether the kidney might be a site of free DNA transport. [32P]-phosphorothioate oligonucleotides (20 mers) were excreted in urine but cleared at only 30% of glomerular filtration rate.

Alterations in glomerular dynamics in congenital, unilateral hydronephrosis.

We have previously shown that rats with congenital, unilateral hydronephrosis exhibit a reduction in GFR that returns to normal when either the renin angiotensin system or thromboxane A2 (TxA2) is blocked. The current study defines the single nephron defect in congenital, unilateral hydronephrosis and evaluates the roles of angiotensin II (Ang II) and TxA2 in this renal derangement. Renal micropuncture experiments were performed on the right kidney of rats from an inbred colony with unilateral right-sided hydronephrosis (HYDRO), or non-affected litter mates (CONTROL).

Upregulation of renin-angiotensin system and downregulation of kallikrein in obstructive nephropathy.

The purpose of this study was to delineate the effects of prolonged (1 and 5 wk) unilateral ureteral obstruction (UUO) on the intrarenal renin-angiotensin and kallikrein-kinin systems in the rat. Systolic blood pressure (SBP) and plasma angiotensin (ANG) II levels were significantly higher at 1 and 5 wk of obstruction than in sham-operated groups. Also, plasma renin activity and ANG I levels were elevated at 1 wk (P < 0.