Fourteen quick tips for crowdsourcing geographically linked data for public health advocacy.

This article presents 14 quick tips to build a team to crowdsource data for public health advocacy. It includes tips around team building and logistics, infrastructure setup, media and industry outreach, and project wrap-up and archival for posterity.

Fluorometric coupled enzyme assay for N-sulfotransferase activity of N-deacetylase/N-sulfotransferase (NDST).

N-Deacetylase/N-sulfotransferases (NDSTs) are critical enzymes in heparan sulfate (HS) biosynthesis. Radioactive labeling assays are the preferred methods to determine the N-sulfotransferase activity of NDST. In this study, we developed a fluorometric coupled enzyme assay that is suitable for the study of enzyme kinetics and inhibitory properties of drug candidates derived from a large-scale in silico screening targeting the sulfotransferase moiety of NDST1. The assay measures recombinant mouse NDST1 (mNDST1) sulfotransferase activity by employing its natural substrate adenosine 3'-phophoadenosine-5'-phosphosulfate (PAPS), a bacterial analog of desulphated human HS, Escherichia coli K5 capsular polysaccharide (K5), the fluorogenic substrate 4-methylumbelliferylsulfate and a double mutant of rat phenol sulfotransferase SULT1A1 K56ER68G. Enzyme kinetic analysis of mNDST1 performed with the coupled assay under steady state conditions at pH 6.8 and 37°C revealed Km (K5) 34.8 µM, Km (PAPS) 10.7 µM, Vmax (K5) 0.53 ± 0.13 nmol/min/µg enzyme, Vmax (PAPS) 0.69 ± 0.05 nmol/min/µg enzyme and the specific enzyme activity of 394 pmol/min/µg enzyme. The pH optimum of mNDST1 is pH 8.2. Our data indicate that mNDST1 is specific for K5 substrate. Finally, we showed that the mNDST1 coupled assay can be utilized to assess potential enzyme inhibitors for drug development.

Mutations in ANAPC1, Encoding a Scaffold Subunit of the Anaphase-Promoting Complex, Cause Rothmund-Thomson Syndrome Type 1.

Rothmund-Thomson syndrome (RTS) is an autosomal-recessive disorder characterized by poikiloderma, sparse hair, short stature, and skeletal anomalies. Type 2 RTS, which is defined by the presence of bi-allelic mutations in RECQL4, is characterized by increased cancer susceptibility and skeletal anomalies, whereas the genetic basis of RTS type 1, which is associated with juvenile cataracts, is unknown. We studied ten individuals, from seven families, who had RTS type 1 and identified a deep intronic splicing mutation of the ANAPC1 gene, a component of the anaphase-promoting complex/cyclosome (APC/C), in all affected individuals, either in the homozygous state or in trans with another mutation. Fibroblast studies showed that the intronic mutation causes the activation of a 95 bp pseudoexon, leading to mRNAs with premature termination codons and nonsense-mediated decay, decreased ANAPC1 protein levels, and prolongation of interphase. Interestingly, mice that were heterozygous for a knockout mutation have an increased incidence of cataracts. Our results demonstrate that deficiency in the APC/C is a cause of RTS type 1 and suggest a possible link between the APC/C and RECQL4 helicase because both proteins are involved in DNA repair and replication.

Standardization and alignment of data capture practices to clinical processes in the evaluation of living kidney donor candidates.

The Living Kidney Donation Program at the Toronto General Hospital, University Health Network sought to develop a comprehensive, secure, accurate, and up-to-date information system for the purposes of quality improvement, research, and performance evaluation. The Comprehensive Living Kidney Donor Database (CLiKeD) houses comprehensive demographic, medical, psychosocial, and evaluation data on living kidney donor candidates abstracted from multiple health information sources. Data are routinely audited to ensure high data quality. Over 3,500 living kidney donor candidates are currently included in CLiKeD. The development of this data system will allow for regular performance assessments of the program, implementation of quality improvement initiatives, and the completion of high-impact, clinically relevant research. In addition, the conception and development of CLiKeD has been instrumental in improving documentation of personal health information at the point of care.