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.     

A transgenic T cell receptor restores thymocyte differentiation in interleukin-7 receptor alpha chain-deficient mice

Traffic accidents are a well-known public health problem worldwide. In Mexico research into risk factors for motor involving vehicles accidents and their consequences has recently been taken into account. The relevant literature does not normally describe the methodological aspects involved in the collection of primary data, since most studies have used secondary data the good quality and validity of which are assumed. The paper presented seeks to discuss and share with researchers in this field, some of the methodological aspects to be considered in the attempt to recreate the scene of the accident and obtain information approximating to reality. The measurements in situ of, such traffic accident variables as injury, use of seat belt, speed and alcohol intake are discussed.     

Shear and extensional flow of reinforced plastics in injection molding. I. Effects of temperature and shear rate with bulk molding compound

Capillary flow studies on bulk molding compound (BMC) using an instrumented injection-molding machine are reported. The significance of extensional flow effects with fiber-reinforced materials is emphasized. The extensional flow behavior in converging dies is modeled, and a means of evaluating both extensional and shear viscosity from capillary flow data is proposed. Methods of correcting results for the effect of deformation heating are discussed. The shear and extensional flow behavior of BMC in the temperature region 18 to 58°C can be fitted to a simplified Arrhenius Law.     

Cellular mechanisms underlying carbachol-induced oscillations of calcium-dependent membrane current in smooth muscle cells from mouse anococcygeus

1. At a holding potential of -40 mV, carbachol (50 microM) produced a complex pattern of inward currents in single smooth muscle cells freshly isolated from the mouse anococcygeus. Membrane currents were monitored by the whole-cell configuration of the patch-clamp technique. Previous work has identified the first, transient component as a calcium-activated chloride current (ICl(Ca)) and the second sustained component as a store depletion-operated non-selective cation current (I(DOC)). The object of the present study was to examine the cellular mechanisms underlying the third component, a series of inward current oscillations (I(oscil)) superimposed on I(DOC). 2. Carbachol-induced I(oscil) (amplitude 97 +/- 11 pA; frequency 0.26 +/- 0.02 Hz) was inhibited by the chloride channel blocker anthracene-9-carboxylic acid (A-9-C; 1 mM), and by inclusion of 1 mM EGTA in the patch-pipette filling solution. 3. In calcium-free extracellular medium (plus 1 mM EGTA), carbachol produced an initial burst of oscillatory current which lasted 94 s before decaying to zero; I(oscil) could be restored by re-admission of calcium. The frequency, but not the amplitude, of I(oscil) increased with increasing concentrations of extracellular calcium (0.5-10 mM). 4. Inclusion of the inositol triphosphate (IP3) receptor antagonist heparin (5 mg ml(-1) in the patch-pipette filling solution, or pretreatment of cells with the sarcoplasmic reticulum (SR) calcium ATPase inhibitor cyclopiazonic acid (CPA; 10 microM), prevented the activation of I(oscil) by carbachol. Caffeine (10 mM) activated both ICl(Ca) and I(DOC) and prevented the induction of I(oscil) by carbachol. Caffeine and CPA also abolished I(oscil) in the presence of carbachol, as did both a low (3 microM) and a high (30 microM) concentration of ryanodine. 5. Carbachol-induced I(oscil) was abolished by the general calcium entry blocker SKF 96365 (10 MM) and by Cd2+ (100 microM), but was unaffected by La3+ (400 microM). As found previously, I(DOC) was also blocked by SKF 96365 and Cd2+, but not La3+; the inhibition of I(DOC) preceded the abolition of I(oscil) by 27 s with SKF 96365 and by 30 s with Cd2+. Nifedipine (1 microM) produced a partial inhibition of the carbachol-induced I(oscil) frequency at holding potentials of -20 mV and -60 mV and, in addition, reduced I(DOC) at -60 mV by 18%. 6. It is concluded that carbachol-induced inward current oscillations in mouse anococcygeus cells are due to a calcium-activated chloride current, and reflect oscillatory changes in cytoplasmic calcium ion concentration. These calcium oscillations are derived primarily from the SR stores, but entry of calcium into the cell is necessary for store replenishment and maintenance of the oscillations. Capacitative calcium entry (via I(DOC) appears to be important not only for sustained contraction of this tissue, but also as a route for re-filling of the SR and, therefore, represents an important target for the development of novel and selective drugs.     

A constrained joint source/channel coder design

The design of joint source/channel coders in situations where there is residual redundancy at the output of the source coder is examined. It has previously been shown that this residual redundancy can be used to provide error protection without a channel coder. In this paper, this approach is extended to conventional source coder/convolutional coder combinations. A family of nonbinary encoders is developed which more efficiently use the residual redundancy in the source coder output. It is shown through simulation results that the proposed systems outperform conventional source-channel coder pairs with gains of greater than 9 dB in the reconstruction SNR at high probability of error     

Langmuir films from antibodies based on amphiphilic polyelectrolytes

A technique for forming Langmuir films from antibodies based on an amphiphilic polyelectrolyte was developed. The physicochemical and immunochemical properties of the Langmuir films obtained were studied. The interaction of HBsAg with the films was found to be described by a model with one binding site, whereas that of HBsAg with antibodies adsorbed on a polystyrene plate, by a model with a positive cooperativity. The use of the novel Langmuir films from antibodies increases the sensitivity of the immunoenzyme assay.     

The surgical treatment of disorders in the formation and fusion of the lumbar vertebrae in children

OBJECTIVE: To investigate the value of polymerase chain reaction (PCR) for follow-up patients infected by Chlamydia trachomatis. METHODS: Follow-up specimens were collected from 30 patients. Chlamydia trachomatis positive were detected by PCR and direct fluorescence assay test (DFA) in the 30 patients before therapy. 15 patients were treated with minocycline (100 mg twice daily) for 10 days, and 15 patients were treated with 1.0 g of azithromycine as a single oral dose. RESULTS: After 1-2 weeks of antimicrobial therapy, all patients had negative DFA for Chlamydia trachomatis, but 9 had positive Chlamydia trachomatis DNA as detected by PCR. CONCLUSIONS: The 9 specimens were not confirmed to livae viable organisms of Chlamydia trachomatis. The debris of nonviable Chlamydia trachomatis DNA was excluded from urinogenital tract at about one month.     

The application of nonlinear structures to the reconstruction ofbinary signals

The problem of reconstructing digital signals which have been passed through a dispersive channel and corrupted with additive noise is discussed. The problems encountered by linear equalizers under adverse conditions on the signal-to-noise ratio and channel phase are described. By considering the equalization problem as a geometric classification problem the authors demonstrate how these difficulties can be overcome by utilizing nonlinear classifiers as channel equalizers. The manner in which neural networks can be utilized as adaptive channel equalizers is described, and simulation results which suggest that the neural network equalizers offer a performance which exceeds that of the linear structures, particularly in the high-noise environment, are presented     

Surface-mediated effects on porous polymer monolith formation within capillaries

The formation of a porous polymer monolith (PPM) is influenced by the physico-chemical properties of the wall surface of its container. This influence can have a dramatic effect on the resulting monolith morphology depending on the nature and composition of the wall. Indeed, a dense polymer layer or “sheath”, distinct from the bulk porous material, has been observed at the wall surface of capillaries, and thus a study was undertaken to explore the dependence of this layer on the hydrophobicity of the surface. A range of silanizing reagents were used to modify the surface of the fused silica capillary, including aminopropyl, trimethylsilyl, octadecyl and perfluorooctyl functionalities. Crosslinked butyl acrylate-based PPM was formed in the modified capillaries and extruded. SEM images of the monoliths were used to examine the sheath morphology and thickness, which are discussed with respect to surface hydrophobicity.