Publications

The protease ADAMTS13 (a disintegrin and metalloprotease with thrombospondin type 1 repeat) cleaves multimers of von Willebrand factor, thus regulating platelet aggregation. ADAMTS13 deficiency leads to the fatal disorder thrombotic thrombocytopenic purpura (TTP). It has been observed that cyclosporin A (CsA) treatment, particularly in transplant patients, may sometimes be linked to the development of TTP. Until now, the reason for such a link was unclear. Here we provide evidence demonstrating that cyclophilin B (CypB) activity plays an important role in the secretion of active ADAMTS13. We found that CsA, an inhibitor of CypB, reduces the secretion of ADAMTS13 and leads to conformational changes in the protein resulting in diminished ADAMTS13 proteolytic activity. A direct, functional interaction between CypB (which possesses peptidyl-prolyl cis-trans isomerase (PPIase) and chaperone functions) and ADAMTS13 is demonstrated using immunoprecipitation and siRNA knockdown of CypB. Finally, CypB knock-out mice were found to have reduced ADAMTS13 levels. Taken together, our findings indicate that cyclophilin-mediated activity is an important factor affecting secretion and activity of ADAMTS13. The large number of proline residues in ADAMTS13 is consistent with the important role of cis-trans isomerization in the proper folding of this protein. These results altogether provide a novel mechanistic explanation for CsA-induced TTP in transplant patients.

PURPOSE

To determine whether radiology reports describe clinically significant carotid arterial stenosis in a consistent format that is actionable by ordering clinicians.

MATERIALS AND METHODS

This study was HIPAA compliant. Informed consent was waived. Institutional review board approval was obtained for this retrospective chart review, which included radiology reports of carotid artery imaging for patients hospitalized with ischemic stroke at 127 Veterans Affairs medical centers in 2006-2007. "Clinically significant results" were defined as results with at least 50% stenosis or at least moderate stenosis, excluding complete occlusion. How often clinically significant results were reported as an exact percentage stenosis (such as 60%), range (such as 50%-69%), or category (such as moderate) was determined. Among results reported as a range, how often the range bracketed clinical thresholds of 50% and 70% (typically used to determine appropriateness of carotid arterial revascularization) was determined.

RESULTS

Among 2675 patients, there were 6618 carotid imaging results, of which 1015 (15%) were considered clinically significant. Among 695 clinically significant results at ultrasonography (US), 348 (50%) were described as a range, and another 314 (45%) were reported as an exact percentage stenosis. Among the 348 clinically significant US results reported as a range, 259 (74%) bracketed the thresholds of 50% or 70%. For magnetic resonance angiographic results, 48% (106 of 221) qualitatively described clinically significant results as a category, 38% (84 of 221) as an exact percentage stenosis, and 14% (31 of 221) as a range.

CONCLUSION

In this national health care system, the manner in which clinically significant carotid arterial stenosis was reported varied widely.

RATIONALE

The function of the P2X(7) nucleotide receptor protects against exacerbation in people with mild-intermittent asthma during viral illnesses, but the impact of disease severity and maintenance therapy has not been studied.

OBJECTIVES

To evaluate the association between P2X(7), asthma exacerbations, and incomplete symptom control in a more diverse population.

METHODS

A matched P2RX7 genetic case-control was performed with samples from Asthma Clinical Research Network trial participants enrolled before July 2006, and P2X(7) pore activity was determined in whole blood samples as an ancillary study to two trials completed subsequently.

MEASUREMENTS AND MAIN RESULTS

A total of 187 exacerbations were studied in 742 subjects, and the change in asthma symptom burden was studied in an additional 110 subjects during a trial of inhaled corticosteroids (ICS) dose optimization. African American carriers of the minor G allele of the rs2230911 loss-of-function single nucleotide polymorphism were more likely to have a history of prednisone use in the previous 12 months, with adjustment for ICS and long-acting β(2)-agonists use (odds ratio, 2.7; 95% confidence interval, 1.2-6.2; P = 0.018). Despite medium-dose ICS, attenuated pore function predicted earlier exacerbations in incompletely controlled patients with moderate asthma (hazard ratio, 3.2; confidence interval, 1.1-9.3; P = 0.033). After establishing control with low-dose ICS in patients with mild asthma, those with attenuated pore function had more asthma symptoms, rescue albuterol use, and FEV(1) reversal (P < 0.001, 0.03, and 0.03, respectively) during the ICS adjustment phase.

CONCLUSIONS

P2X(7) pore function protects against exacerbations of asthma and loss of control, independent of baseline severity and the maintenance therapy.

MicroRNAs (miRNAs) are essential components of gene regulation, but identification of miRNA targets remains a major challenge. Most target prediction and discovery relies on perfect complementarity of the miRNA seed to the 3' untranslated region (UTR). However, it is unclear to what extent miRNAs target sites without seed matches. Here, we performed a transcriptome-wide identification of the endogenous targets of a single miRNA-miR-155-in a genetically controlled manner. We found that approximately 40% of miR-155-dependent Argonaute binding occurs at sites without perfect seed matches. The majority of these noncanonical sites feature extensive complementarity to the miRNA seed with one mismatch. These noncanonical sites confer regulation of gene expression, albeit less potently than canonical sites. Thus, noncanonical miRNA binding sites are widespread, often contain seed-like motifs, and can regulate gene expression, generating a continuum of targeting and regulation.