Multicolor Flow Cytometry and Cytokine Analysis Provides Enhanced Information on Kidney Transplant Biopsies.

Introduction: Current processing of renal biopsy samples provides limited information about immune mechanisms causing kidney injury and disease activity. We used flow cytometry with transplanted kidney biopsy samples to provide more information on the immune status of the kidney.

Methods: To enhance the information available from a biopsy, we developed a technique for reducing a fraction of a renal biopsy sample to single cells for multicolor flow cytometry and quantitation of secreted cytokines present within the biopsy sample.

A Novel Three-Dimensional Human Peritubular Microvascular System.

Human kidney peritubular capillaries are particularly susceptible to injury, resulting in dysregulated angiogenesis, capillary rarefaction and regression, and progressive loss of kidney function. However, little is known about the structure and function of human kidney microvasculature. Here, we isolated, purified, and characterized human kidney peritubular microvascular endothelial cells (HKMECs) and reconstituted a three-dimensional human kidney microvasculature in a flow-directed microphysiologic system.

CMS-required visits adversely impact Seattle area hemodialysis program.

Chronic hemodialysis in Seattle developed with a highly patient-centric approach, providing patients the option of dialyzing in neighborhood units under the administration of a non-profit organization, the Northwest Kidney Center (NKC). This study examined the effects of a requirement in the 2009 Conditions for Coverage that in-center hemodialysis patients be visited quarterly during their treatment session. A retrospective analysis of quality outcome indicators for patients managed at the University of Washington Medical Center and dialyzing at NKC in 2008 and 2010 found that dialysis adequacy (eKt/V), phosphorus, albumin, and hemoglobin were not improved by quarterly dialysis visits.

Proliferative potential of human kidney endothelial cells: bone marrow-derived cells may not be required for high proliferation.

Background: Proliferative potential of a single cell, defined as the number of progeny it gives rise to, has been used to define a hierarchy of endothelial progenitor cells in blood. Cells with high proliferative potential are presumed to have greater capacity for endothelium repair. Based on results with commercially available endothelial cells, it has been proposed that a proliferative hierarchy of endothelial cells also exists within blood vessels.

Reoperative thyroidectomy for benign thyroid disease.

Background: Subtotal thyroidectomy for benign thyroid disease (BTD) may lead to delayed recurrence, thus necessitating reoperative surgery. We describe our experience with reoperative thyroidectomy for BTD and recommendations for definitive primary management.

Methods: Patients undergoing thyroid surgery between 2003 and 2007 by a single surgeon were prospectively assessed.

Findings from a public thyroid screening protocol: ultrasound and disease characteristics.

Objectives: A public thyroid screening protocol incorporating ultrasonography was developed and implemented as a feature of the National Thyroid Cancer Awareness month. Findings and lessons learned are described.

Methods: Prospective analysis of participants in a 1-day thyroid screening protocol and review of findings and referrals generated during the screening process.

Noninvasive measurement of protein aggregation by mutant huntingtin fragments or alpha-synuclein in the lens.

Many diverse human diseases are associated with protein aggregation in ordered fibrillar structures called amyloid. Amyloid formation may mediate aberrant protein interactions that culminate in neurodegeneration in Alzheimer, Huntington, and Parkinson diseases and in prion encephalopathies. Studies of protein aggregation in the brain are hampered by limitations in imaging techniques and often require invasive methods that can only be performed postmortem.

Laryngeal nerve monitoring and minimally invasive thyroid surgery: complementary technologies.

Objective: To determine the feasibility of the combined use of laryngeal nerve monitoring and minimally invasive thyroid surgery.

Design: Prospective, nonrandomized analysis of single-surgeon experience.

Setting: Academic institution.

Senataxin, the yeast Sen1p orthologue: characterization of a unique protein in which recessive mutations cause ataxia and dominant mutations cause motor neuron disease.

A severe recessive cerebellar ataxia, Ataxia-Oculomotor Apraxia 2 (AOA2) and a juvenile onset form of dominant amyotrophic lateral sclerosis (ALS4) result from mutations of the Senataxin (SETX) gene. To begin characterization this disease protein, we developed a specific antibody to the DNA/RNA helicase domain of SETX. In murine brain, SETX concentrates in several regions, including cerebellum, hippocampus and olfactory bulb with a general neuronal expression profile, colocalizing with NeuN.

SIMPLE interacts with NEDD4 and TSG101: evidence for a role in lysosomal sorting and implications for Charcot-Marie-Tooth disease.

Mutation of the SIMPLE gene (small integral membrane protein of the lysosome/late endosome) is the molecular basis of Charcot-Marie-Tooth disease type 1C (CMT1C), a demyelinating peripheral neuropathy. Although the precise function of SIMPLE is unknown, prior reports suggest it localizes to the lysosome/late endosome. Furthermore, murine Simple interacts with Nedd4 (neural precursor cell expressed, developmentally downregulated 4), an E3 ubiquitin ligase that is important for regulating lysosomal degradation of plasma membrane proteins.

Normal human kidney HLA-DR-expressing renal microvascular endothelial cells: characterization, isolation, and regulation of MHC class II expression.

Human, but not murine, renal peritubular and glomerular capillaries constitutively express class II major histocompatibility (MHC) proteins at high levels in normal human kidney. Expression of class II proteins on renal microvascular endothelial cells (RMEC) makes it available to circulating lymphocytes and imparts a surveillance capacity to RMEC for controlling inflammatory responses. In this report, the co-expression of HLA-DR and the endothelial marker CD31 are used to identify RMEC as a distinct population of cells within a standard renal biopsy using flow cytometry.