Of the various conservative treatment modalities available for temporomandibular disorders, we believe that therapeutic exercise has a good prognosis, especially for anterior disc displacement without reduction. Since its effectiveness has not been extensively evaluated, we conducted a comparative study to verify the hypothesis that treatment efficacy would not differ for exercise and occlusal splints. Fifty-two individuals with anterior disc displacement without reduction were randomly assigned to a splint or a joint mobilization self-exercise treatment group. Four outcome variables were evaluated: (i) maximum mouth-opening range without and (ii) with pain, (iii) current maximum daily pain intensity, and (iv) limitation of daily functions. All outcome variables significantly improved after 8 weeks of treatment in both groups. In particular, the mouth opening range increased more in the exercise group than in the splint group. This result demonstrates that therapeutic exercise brings earlier recovery of jaw function compared with splints.

Recombinant pro‐Der p 1 expressed in yeast Pichia pastoris was convertible into the prosequence‐removed mature Der p 1 with full activities of cysteine protease and IgE‐binding with or without N‐glycosylation of the mature sequence as well as pro‐Der f 1. The active recombinant variants will be the basis for various future studies. The major N‐terminus of pro‐Der p 1 with low proteolytic activity was the putative signal‐cleavage site, while that of pro‐Der f 1 contained not only the equivalent site but also 21 residues downstream, and pro‐Der f 1 retained significant activity. Contribution of the N‐terminal region of the Der p 1 prosequence including an N‐glycosylation motif on effective inhibition of proteolytic activity of pro‐Der p 1 was suggested.

There are various antigenic variants of Orientia tsutsugamushi which are distinguished by immunological and molecular genetic methods targeted at the antigenic diversity of 56-kDa type-specific antigen proteins. The present study was performed to analyze 15 strains successfully isolated from rodents in Saitama Prefecture, Japan, by 56-kDa gene sequence homologies, reactivities with type-specific monoclonal antibodies and polymerase chain reaction (PCR) using type-specific primer-pairs. We demonstratedthe presence of a new type of O. tsutsugamushi among the isolates. This new type, designated as the Saitama type, was located in the branch of Karp type in the phylogenetic tree based on 56-kDa gene sequences, but distant from the known Karp types, such as Karp, JP-l and JP-2, showing less than 90% homology. Strains of this type could not be distinguished by immunological methods from Karp type strains, but a new primer-pair for PCR which specifically amplifies the DNA of this new type strain was designed. This primerpair may serve to find this strain type in future studies.

Abstract

Background: Structurally refolded recombinant forms of major house dust mite group 2 allergens, Der f 2 and Der p 2, expressed in Escherichia coli, were prepared by solubilizing the insoluble products with urea and subsequently dialyzing against buffer. In this study, we determined conditions for refolding the urea-denatured recombinant Der f 2 and Der p 2 by one-step dilution as an alternative to dialysis, which requires several steps of handling and much time and cost. Methods: The insoluble bacterial product containing recombinant Der f 2 was solubilized with a buffer containing 8 M urea, and the solution was diluted to various urea concentrations. The refolding efficiency in each dilution was estimated from the height of the peak corresponding to the folded recombinant Der f 2 and that containing the aggregated form on anion exchange chromatography. The structure and allergenicity of the purified recombinant Der f 2 and Der p 2 refolded using the dilution method were analyzed based on circular dichroism and a basophil histamine-releasing assay, respectively. Results: Although the refolding efficiency decreased as the urea concentration in the dilution increased, experimental conditions whereby the protein and urea concentrations in the dilution were less than 0.5 mg/ml and 0.8 M, respectively, achieved maximum refolding efficiency. The recombinant allergens prepared by the dilution method exhibited the secondary structure and histamine-releasing activity of natural allergens purified from mite culture. Conclusions: The dilution method established in this study is more convenient in terms of handling, time, and cost than the dialysis method and will be useful for large-scale production and for the preparation of numbers of mutants to analyze IgE epitopes.