The OKT3 Antibody Response Study: a multicentre study of human anti-mouse antibody (HAMA) production following OKT3 use in solid organ transplantation

Human anti-murine antibody titres following patient exposure to the monoclonal antibody Orthoclone OKT3 (muromonab-CD3) are determined by laboratories using diverse analytical methods which are not standardized and whose concordance is not established. A multicentre study group therefore compared testing for IgG anti-OKT3 antibody among seven laboratories. A set of 270 sera was obtained from 30 heart, 30 kidney and 30 liver transplant recipients with no previous exposure to OKT3 who were receiving OKT3 for induction immunosuppression. Sera were collected from each patient prior to and at 24 +/- 2 days and 31 +/- 2 days following initial OKT3 exposure. Identical aliquots of all 270 sera were tested for IgG anti-OKT3 antibody by each laboratory. In addition, the limit of detection of each laboratory's method was estimated by titration of an affinity-purified IgG anti-OKT3 reference material of known concentration. Anti-OKT3 antibody formation differed greatly among the three organ groups. Cardiac patients demonstrated the least sensitization and almost exclusively lower titres, while kidney recipients had more frequent and higher titre antibody formation. Liver recipients yielded the highest sensitization rate and the most frequent high titre sera. Importantly, the seven laboratories differed widely in the number of pretreatment sera reported as positive (ranging from 0% to 41% among laboratories), the number of post-OKT3 sera reported as positive (17-63%), the number of post-OKT3 samples with titre > or = 1000 (2-31%), and the number of patients sensitized 19-69%). Concordance among laboratories was highly variable, with interlaboratory agreement ranging from 38% to 83% on the sample titres assigned to 180 post-OKT3 sera. Many of the discordant results were consistent with differences in the limit of detection of the analytical methods, which ranged from 0.19 microgram/ml to > or = 15 micrograms/ml, a nearly 100-fold difference among laboratories. This study demonstrated the presence of both good concordance and significant discordance among laboratories in determining human anti-mouse antibody titres, and demonstrated that common titre categories (100, 1000, 10,000) were not equivalent among laboratories. The level of concordance among methods should be considered when comparing anti-OKT3 antibody results from different centres and their correlation with clinical events. Universal comparative testing, patterned after proficiency testing programmes, is needed to assess differences among laboratories and to bring uniformity and a sound interpretative basis to this field of testing.     

Deoxyribonucleic acid vaccines encoding antigens with rapid proteasome-dependent degradation are highly efficient inducers of cytolytic T lymphocytes

We generated plasmid expression vectors encoding ubiquitin and beta-galactosidase (beta-gal) with different intervening amino acids, allowing for the production of processed protein products that have either stabilizing or destabilizing residues at their N-termini. P815 cells transfected with plasmids encoding beta-gal with a destabilizing N-terminus did not have detectable expression beta-gal unless they were treated with inhibitors specific for the proteasome. Inhibitors of other proteolysis pathways had no such effect. Nevertheless, transfectants expressing beta-gal with different amino acid residues were equally sensitive to cytolysis by a CTL clone specific for a beta-gal peptide presented in the context of H-2Ld. In contrast to vectors encoding native beta-gal, plasmid vectors encoding beta-gal with a destabilizing residue did not induce detectable anti-beta-gal Abs when injected into skeletal muscle of BALB/c mice. However, such vectors were significantly more effective than vectors encoding native beta-gal or beta-gal with a stabilizing residue in stimulating CTL specific for P13.2, a lacZ transfectant of P815. We conclude that incorporation of strategies that enhance proteasome-dependent degradation may generate DNA vaccines that are more effective in inducing cellular immunity against targeted Ags.     

A new tool to measure pressure under burn garments

This article introduces a new tool to measure the pressure that is under pressure garments. The Iscan (Tekscan, Inc.) system uses a patented ultra-thin (0.007 inch) sensor with multiple sensing locations that sample continuously at 100 times per second. It is noninvasive, convenient, and quick. The study had two parts. First, we established the validity and reliability of the device. Next, garment/scar interface pressures were measured on new garments with use of the Iscan system. Four garment types were studied, with 10 measurements made in each group: Isotoner gloves (Smith & Nephew Roylan, Inc.); custom-fit pressure gloves; Tubigrip forearm sleeves (Seton Health Care Group); and custom-fit pressure forearm sleeves. Mean garment/scar interface pressures were 18 +/- 2 mm Hg for the Isotoner glove, 34 +/- 5 mm Hg for the custom-fit pressure glove, 20 +/- 7 mm Hg for the Tubigrip sleeve, and 35 +/- 6 mm Hg for the custom-fit sleeve. We concluded that the Iscan system can be used to measure pressure under pressure garments accurately and reliably, and that custom-fit hand and forearm garments provide more pressure than Isotoner gloves or Tubigrip sleeves.     

Porphyromonas (Bacteroides) endodontalis: its role in endodontal infections

Porphyromonas endodontalis (formerly Bacteroides endodontalis) is a black-pigmented anaerobic Gram-negative rod which is associated with endodontal infections. It has been isolated from infected dental root canals and submucous abscesses of endodontal origin. The presence of P. endodontalis in infected dental root canals has been correlated with symptoms of an acute infection. It is occasionally found on oral mucous membranes and periodontal pockets. P. endodontalis has shown relatively low virulence in experimental monoinfections. In anaerobic mixed infections it can play an essential role. Differences in virulence between strains have been related to capsular material. On the basis of different types of capsules, three serotypes have been described. P. endodontalis is sensitive to a wide range of antibiotics, including the penicillins, the tetracyclines, and metronidazole.     

A set of electrophoretic molecular markers for strain typing and population genetic studies of Histoplasma capsulatum

A set of eleven biallelic and three multiallelic molecular markers have been developed to analyze populations of Histoplasma capsulatum. All markers are amplified by polymerase chain reaction (PCR) and can be readily scored using minimal amounts of template DNA. The 11 biallelic loci have polymorphic restriction endonuclease sites or small insertions or deletions which may be assessed by agarose gel electrophoresis. These markers are inherited in an unambiguous manner and are ideal for assessing structure and gene flow within US populations of H. capsulatum, but are monomorphic in non-US populations. Both length and sequence variation are present in the multiallelic loci, which can be scored by direct sequencing, polyacrylamide gel electrophoresis, or single-strand conformation polymorphism (SSCP): As they are hypervariable, the multiallelic loci can be used to type isolates and to assess the level of genetic variation within populations. Preliminary results indicate that the three multiallelic markers presented are sufficient to distinguish isolates at the individual level and are polymorphic in both US and non-US populations. This collection of molecular markers will be a useful tool in population and epidemiology studies of H. capsulatum.     

Economics of liquid hydrogen from water electrolysis

Using the data from vendors and open literature, we have constructed a preliminary cost model to predict the cost of liquid hydrogen (LH₂) from water electrolysis. The model has been applied to two LH₂ plants. Both have a production rate of 10 886 kg day⁻¹. The first plant (Plant 1) consists of a tank-type electrolyzer and a liquefier and runs all day at the base-load (or equivalent base-load) power rate. The second (Plant 2) consists of a tank-type electrolyzer, a liquefier, a compressor, and storage vessels. The electrolyzer and the compressor shall run only during the off-peak hours at the off-peak power rate. However, the liquefier shall run all day at the base-load (or equivalent base-load) power rate. The costs of LH₂ from both plants are presented. Unless the base-load power rate is high and the off-peak power rate is substantially low, Plant 1 appears to be economically more feasible than Plant 2.     

Mechanical properties of poly(ether-ether-ketone) for engineering applications

An outline of the characteristics of PEEK and the versatility of its compositional forms (micro and macro composites) are given to illustrate its wide potential for success in engineering applications. Although it is necessary to have particular tabulations of mechanical properties for engineering design, these are seldom available and consequently it is argued that an understanding of stiffness, toughness and strength properties are required to fully exploit available manufacturer's data and thus develop the full potential of PEEK and its composites. Stiffness characteristics are considered in terms of a modulus function which is dependent on time under load and temperature. In its composite forms, whether reinforced with short or continuous fibres, stiffness anistropy can be both considerable and complex, but some empirical ground-rules are apparent. For continuous fibre composites even in the form of complex lay-ups, it is also possible to attempt some stiffness prediction from certain pseudo-elastic constants. Toughness of PEEK and its composites is described in terms of both comparative and intrinsic properties. Instrumented falling weight impact data, particularly as a function of temperature enable some insight into ductile-brittle transitions for the unreinforced material, but crack initiation and crack propagation processes for the various fibre reinforced forms. Intrinsic toughness is described in terms of linear elastic fracture mechanics theory. Strength properties are described for static and dynamic loading configurations. In particular, PEEK and its composites are evaluated for increasing test severities for strength characteristics; stress concentration, loading form and test temperature are considered.