Identification of potent and selective RNA antagonists of the IFN-gamma-inducible CXCL10 chemokine.

CXCL10 (also known as IP-10 in humans and CRG-2 in mice) is a nonglycosylated chemokine and a member of the non-ELR CXC chemokine subfamily implicated in a variety of inflammatory conditions. The role of CXCL10 in different disease states still requires clarification, and new approaches are necessary to better understand its biological function. We report here the isolation of a series of nuclease-resistant RNA aptamers that act to antagonize human CXCL10 function in a number of in vitro and cell-based assays. The two most potent aptamers identified were highly selective for human CXCL10. A further aptamer was identified that antagonized both the human and the mouse CXCL10. A combination of a molecular-biology-based truncation and solid-phase synthesis enabled the truncation of one of the aptamers from 71 to 34 nucleotides. This was followed by PEGylation, 3' capping, and further stabilization of the RNA aptamer, while its high potency was maintained. These aptamers could be utilized as powerful target validation tools and may also have therapeutic potential. To our knowledge, the CXCL10 aptamers generated are the most potent antagonists of CXCL10/CXCR3 signaling reported to date.

A comparative reliability analysis of computer-generated bitemark overlays.

This study compared the reliability of two methods used to produce computer-generated bitemark overlays with Adobe Photoshop (Adobe Systems Inc., San Jose, CA). Scanned images of twelve dental casts were sent to 30 examiners with different experience levels. Examiners were instructed to produce an overlay for each cast image based on the instructions provided for the two techniques. Measurements of the area and the x-y coordinate position of the biting edges of the anterior teeth were obtained using Scion Image software program (Scion Corporation, Frederick, MD) for each overlay. The inter- and intra-reliability assessment of the measurements was performed using an analysis of variance and calculation of reliability coefficients. The assessment of the area measurements showed significant variances seen in the examiner variable for both techniques resulting in low reliability coefficients. Conversely, the results for the positional measurements showed no significant differences in the variances between examiners with exceptionally high reliability coefficients. It was concluded that both techniques were reliable methods to produce bitemark overlays in assessing tooth position.

11-Dehydro-thromboxane B2, a stable thromboxane metabolite, is a full agonist of chemoattractant receptor-homologous molecule expressed on TH2 cells (CRTH2) in human eosinophils and basophils.

Thromboxane (TX) A(2), a cyclooxygenase-derived mediator involved in allergic responses, is rapidly converted in vivo to a stable metabolite, 11-dehydro-TXB(2), which is considered to be biologically inactive. In this study, we found that 11-dehydro-TXB(2), but not the TXA(2) analogue U46,619 or TXB(2), activated eosinophils and basophils, as assayed by flow cytometric shape change. 11-Dehydro-TXB(2) was also chemotactic for eosinophils but did not induce, nor inhibit, platelet aggregation. Chemoattractant receptor-homologous molecule expressed on TH2 cells (CRTH2) is an important chemoattractant receptor expressed by eosinophils, basophils, and TH2 lymphocytes, and prostaglandin (PG)D(2) has been shown to be its principal ligand. 11-Dehydro-TXB(2) induced calcium flux mainly from intracellular stores in eosinophils, and this response was desensitized after stimulation with PGD(2) but not other eosinophil chemoattractants. Shape change responses of eosinophils and basophils to 11-dehydro-TXB(2) were inhibited by the thromboxane (TP)/CRTH2 receptor antagonist ramatroban, but not the selective TP antagonist SQ29,548, and were insensitive to pertussis toxin. The phospholipase C inhibitor U73,122 attenuated both 11-dehydro-TXB(2)- and PGD(2)-induced shape change. 11-Dehydro-TXB(2) also induced the chemotaxis of BaF/3 cells transfected with hCRTH2 but not naive BaF/3 cells. At a threshold concentration, 11-dehydro-TXB(2) had no antagonistic effect on CRTH2-mediated responses as induced by PGD2. These data show that 11-dehydro-TXB(2) is a full agonist of the CRTH2 receptor and hence might cause CRTH2 activation in cellular contexts where PGD-synthase is not present. Given its production in the allergic lung, antagonism of the 11-dehydro-TXB(2)/CRTH2axis may be of therapeutic relevance.

Adherence of forensic odontologists to the ABFO guidelines for victim evidence collection.

Certifying boards for different professions have the duty to help establish standards and guidelines for methodologies routinely performed within the discipline. For forensic dentists, this responsibility is placed upon the American Board of Forensic Odontology (ABFO). The purpose of this study was to examine whether board certified and noncertified forensic odontologists adhere to the ABFO Guidelines outlined in the collection of victim bitemark evidence. A questionnaire was developed to assess the compliance and attitudes towards the typical evidence collected, the photographic documentation, and the handling of the bite site injury. The results indicate the majority of the respondents in both representative groups routinely follow the guidelines set forth by the ABFO. The lack of personally photographing the bite injury on a consistent basis is an area of concern for all examiners. The photographic evidence is an instrumental part of the investigation and often cannot be utilized due to improper procedures being followed. The film type utilized, bite site impression techniques, and excision of any tissue samples remain an individual choice and vary significantly among each forensic odontologist.