GDF-5 deficiency in mice delays Achilles tendon healing.

The aim of this study was to examine the role of one of the growth/differentiation factors, GDF-5, in the process of tendon healing. Specifically, we tested the hypothesis that GDF-5 deficiency in mice would result in delayed Achilles tendon repair. Using histologic, biochemical, and ultrastructural analyses, we demonstrate that Achilles tendons from 8-week-old male GDF-5 -/- mice exhibit a short-term delay of 1-2 weeks in the healing process compared to phenotypically normal control littermates. Mutant animals took longer to achieve peak cell density, glycosaminoglycan content, and collagen content in the repair tissue, and the time course of changes in collagen fibril size was also delayed. Revascularization was delayed in the mutant mice by 1 week. GDF-5 deficient Achilles tendons also contained significantly more fat within the repair tissue at all time points examined, and was significantly weaker than control tissue at 5 weeks after surgery, but strength differences were no longer detectable by 12-weeks. Together, these data support the hypothesis that GDF-5 may play an important role in modulating tendon repair, and are consistent with previously posited roles for GDF-5 in cell recruitment, migration/adhesion, differentiation, proliferation, and angiogenesis.

Advances in the chemical synthesis and purification of RNA.

Significantly developments that improve the chemical synthesis and purification of oligoribonucleotides have been attained. Introduction of a 2'-O-acetyl function on the nucleoside bound to the polystyrene support enhanced the yield of the oligoribonucleotide after cleavage. Higher coupling efficiency was achieved by the activation of phosphoramidites with a 0.75M solution of 5-ethylthio-1 H-tetrazole. Removal of the 2'-O-t-butyldimethylsilyl groups was effected rapidly at elevated temperatures with triethylamine trihydrofluoride in DMF. Fully deprotected RNA was then directly desalted and precipitated by the addition of 1-butanol. Purified RNA was isolated by the addition of 1-propanol to the collected HPLC purified fractions thus avoiding laborious desalting and solvent evaporation processes. These advancements have been extended to synthesis of ribozymes and tRNA.

Vertical distortion in distal extension ridges and palatal area of casts made by different techniques.

A coordinate measurement machine with laser probe was used to measure the vertical distortion of the casts produced by use of three types of impression materials (irreversible hydrocolloid, condensation silicone, and addition silicone) and two types of trays (stock and custom trays). Results indicated that all impression groups showed positive vertical distortion (ranging from 0.00566 to 0.30299 mm) at the edentulous ridges and palatal area. The amount of the vertical distortion was greatest at the palatal area and was followed by the high edentulous ridge and the low edentulous ridge. Addition silicone, with either custom tray or stock tray, was the most accurate impression material. Condensation silicone was more accurate than irreversible hydrocolloid in custom tray impression. However, in stock tray impression the irreversible hydrocolloid was more accurate than the condensation silicone. The results suggest that, with careful manipulation, irreversible hydrocolloid with stock tray impression may provide a satisfactory cast for fabricating the framework of a distal extension removable partial denture.

Structural RNA mimetics: N3'-->P5' phosphoramidate DNA analogs of HIV-1 RRE and TAR RNA form A-type helices that bind specifically to Rev and Tat-related peptides.

An attractive strategy for the development of anti-retroviral drugs is the exploration of compounds that mimic RNA control regions of the viral genome and act as "decoys" to sequester viral gene regulatory proteins. Decoys consisting of RNA, however, are chemically unstable and readily degraded by cellular nucleases. DNA decoys, which are slightly more stable, also might not be appropriate because of possible structural differences between RNA and DNA helices and the complexes they form with proteins. It was recently reported, however, that DNA analogs with modified N3'-->P5' phosphoramidate sugar-phosphate backbones are stable and nuclease-resistant and exist predominately as A-form helices in solution [Gryaznov, S., et al. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 5798-5802]. We now report that oligonucleotide N3'-->P5' phosphoramidates DNA analogs of HIV-1 RRE IIB and TAR RNA form stable duplexes that exist in the A form as judged by circular dichroism (CD). Moreover, gel shift assays demonstrate that these phosphoramidates can specifically bind to peptides derived from HIV-1 Rev and Tat proteins. Isosequential phosphodiester DNA duplexes, existing in the B form by CD, do not bind to the respective peptides under the experimental conditions used. These results suggest the possibility that nuclease-resistant oligonucleotide N3'-->P5' phosphoramidates might serve as RNA-like decoys and disrupt specific viral RNA/protein interactions such as RRE/Rev and TAR/Tat in HIV-1.