Citations

Abstract Whole genome amplification (WGA) offers a means to enrich DNA quantities for epidemiologic studies. We used an ovarian cancer study of 1,536 single nucleotide polymorphisms (SNPs) and 2,368 samples to assess performance of multiple displacement amplification (MDA) WGA using an Illumina GoldenGate BeadArray. Initial screening revealed successful genotyping for 93.4% of WGA samples and 99.3% of genomic samples, and 93.2% of SNPs for WGA samples and 96.3% of SNPs for genomic samples. SNP failure was predicted by Illumina-provided designability rank, %GC (P ≤ 0.002), and for WGA only, distance to telomere and Illumina-provided SNP score (P ≤ 0.002). Distance to telomere and %GC were highly correlated; adjustment for %GC removed the association between distance to telomere and SNP failure. Although universally high, per-SNP call rates were related to designability rank, SNP score, %GC, minor allele frequency, distance to telomere (P ≤ 0.01), and, for WGA only, Illumina-provided validation class (P < 0.001). We found excellent concordance generally (>99.0%) among 124 WGA:genomic replicates, 15 WGA replicates, 88 replicate aliquots of the same WGA preparation, and 25 genomic replicates. Where there was discordance, it was across WGA:genomic replicates but limited to only a few samples among other replicates suggesting the introduction of error. Designability rank and SNP score correlated with WGA:genomic concordance (P < 0.001). In summary, use of MDA WGA DNA is feasible; however, caution is warranted regarding SNP selection and analysis. We recommend that biological SNP characteristics, notably distance to telomere and GC content (<50% GC recommended), as well as Illumina-provided metrics be considered in the creation of GoldenGate assays using MDA WGA DNA. (Cancer Epidemiol Biomarkers Prev 2008;17(7):1781–9)

The American Heart Association’s (AHA’s) mission is “to build healthier lives, free of cardiovascular diseases and stroke.” This first article in a 2-part series will serve to present an overview of the work the AHA has undertaken to translate evidence into practice for healthcare professionals. It describes the extensive work of the AHA to support and further the delivery of evidence-based medicine, which includes the following: (1) supporting scientific discovery and the next generation of healthcare professionals and researchers; (2) disseminating scientific information; (3) developing evidence-based guidelines and statements; (4) creating and advocating for the implementation of performance indicators/measures; (5) developing clinical decision support and quality improvement tools; and (6) developing directed-cause campaigns, all of which can lead to improved patient care. This article also discusses the need for novel approaches and some of the AHA’s evolving strategies to help address gaps in care. The second article, which will be published shortly after this one, will examine the AHA’s efforts to engage and empower healthcare consumers to become more involved with their own health and health care.

Abstract Background: The accurate and precise measurement of urinary albumin is critical, since even minor increases are diagnostically sensitive indicators of renal disease, cardiovascular events, and risk for death. To gain insights into potential measurement biases, we systematically compared urine albumin measurements performed by LC-MS, a clinically available immunoturbidimetric assay, and size-exclusion HPLC. Methods: We obtained unused clinical urine samples from 150 patients who were stratified by degrees of albuminuria (<20 mg/L, 20–250 mg/L, >250 mg/L) as determined by the immunoturbidimetric assay used in our clinical laboratory (Roche Hitachi 912). Urine albumin was then remeasured via LC-MS and HPLC (Accumin™) assays. Results: The immunoturbidimetric assay, calibrated using manufacturer-supplied serum-derived calibrators (Diasorin), underestimated albumin compared with LC-MS. After calibration with purified HSA, this immunoturbidimetric assay correlated well with LC-MS. HPLC overestimated albumin compared with both LC-MS and immunoturbidimetry. The current LC-MS and HPLC assays both performed poorly at concentrations <20 mg/L. Conclusions: Efforts are needed to establish gold-standard traceable calibrators for clinical assays. LC-MS is a specific method to quantify albumin in native urine when concentrations exceed 20 mg/L, and therefore could be employed for standardization among assays.

Natural and synthetic hydrogels have attracted much attention for nerve regeneration. Previous studies have shown that electrical charge has significant effect on stimulation of neurite outgrowth. In this work, incorporation of a positively charged monomer into the photocrosslinkable oligo(polyethylene glycol) fumarate (OPF) hydrogel has been investigated. We have evaluated the effect of localized positive charge on neurite outgrowth in culture with an objective that positively charged hydrogels ultimately can be used for stimulating in vivo nerve regeneration. The effect of charged modification has been also studied on mechanical properties and swelling ratio of these hydrogels. Our data indicated that with increasing charge density hydrogels swelling ratio increased in water, however it remained constant in PBS. We also demonstrated that compressive modulus and tensile strength of the hydrogels improved with incorporation of electrical charge into the hydrogels. Biodegradation of modified hydrogels was investigated in a series of biomimetic solutions. OPF hydrogels appeared to be more susceptible to oxidative degradation as opposed to the hydrolytic degradation in enzymes and acidic solution, and the degradation rate was correlated to the PEG molecular weight and charge density of the hydrogels. To investigate the effect of charge modification on nerve cell attachment and differentiation, dissociated dorsal root ganglion (DRG) cells were plated onto the modified and unmodified hydrogels surfaces. DRG cells attached and extended their neurites more readily on the surface of positively charged hydrogels as opposed to the unmodified hydrogels.

The number of osteoporotic hip fractures is projected to increase two to three fold in the next 30years. This will lead to increased mortality and nursing home placement with significant costs to our society. Although in clinical trials drug and hormone therapies have shown an important decrease in fracture risk there is a significant role of surgical implantation treatments to further reduce the risk in specific subpopulations. Strength testing was completed on four pairs of osteoporotic, two pairs of osteopenic, and two pairs of normal cadaver femurs. The fracturing was recorded with a video camera at 4000 frames per second. The specially designed test fixture was instrumented with three load cells to measure the femoral forces. Force and moment data were recorded simultaneously and correlated with the frames. The fracture force at the greater trochanter was used to characterize the strength of implanted∕nonimplanted osteoporotic femurs. All femurs began fracturing within about 60ms at a test speed of 100mm∕s. During the first 20ms, the relationship between force and displacement was nonlinear, but linear thereafter until the fracture occurs. The fracture events resolved completely within 10–20ms. The average strength increase of the implanted femurs was 79% compared to the nonimplanted controls. The simultaneous acquisition of fracture images and load data is a powerful tool to understand and characterize numerically the femoral fracture. This methodology allows the researcher to define the best metrics to compare competing reinforcement materials, implant designs and surgical procedures.

Temporomandibular joint disorder afflicts 10 million Americans, many of whom have osteoarthritis of the temporomandibular joint (TMJ). This condition can inflict severe pain and disrupt the lives of sufferers in many ways. Partial or total replacement of the temporomandibular joint is a last resort treatment option. Surgeons at Mayo Clinic believe a new hemijoint implant design coupled with unique surgical technique can improve joint kinematics and reduce pain. They are currently investigating a patent-pending implant design in a series of patient trials. The Division of Engineering at Mayo Clinic has developed a novel process for fabricating TMJ implants for this study. Computed Tomagraphy (CT) images of the surgical site are first converted into a 3D computer model of the mandibular fossa and condyle area. A fused deposition modeling process is used to create a plastic model of the anatomy, and the surgeons use that model to create a wax mold of the implant. The wax mold is laser scanned to create a 3D CAD model that can be machined with a standard four axis milling machine out of implant grade CoCrMo material. Because of the unique shape of the implant, the machining takes place in two phases, with the implant being refixtured between machining phases using a high strength industrial adhesive. Finally, the implant is polished, inspected, passivated and sterilized for surgery. This fabrication process has allowed Mayo Clinic surgeons to quickly and accurately test their unique implant design.