Monoclonal antibody with hemagglutination, immobilization, and neutralization activities defines an immunodominant, 47,000 mol wt, surface-exposed immunogen of Treponema pallidum (Nichols).

Radioimmunoprecipitation (RIP) analyses performed on 125I-surface-labeled Treponema pallidum cells using various immune sera revealed the presence of six major surface antigens (immunogens) with apparent molecular weights of 47 K, 36 K, 34 K, 32 K, 29 K, and 13 K. Among these, the 47 K surface antigen was most abundant. Radioimmunoprecipitation assays using 125I-labeled T. phagedenis biotype Reiter or immunoblot analyses using the same strain, failed to reveal the presence of the 47 K mol wt antigen in the representative nonpathogenic treponeme. Preabsorption of anti-T. pallidum immune rabbit serum (IRS) with the Reiter organism did not remove anti-T. pallidum antibodies from immune serum that reacted with the 47 K mol wt immunogen or other immunogens of T. pallidum present in the characteristic antigenic profile. Monoclonal antibodies (mAb) directed specifically against the 47 K mol wt immunogen of T. pallidum also failed to react with an analogous 47 K mol wt component in Treponema phagedenis biotype Reiter, further suggesting the unique presence of this antigen in pathogenic treponemes. The presence of the 47 K mol wt surface immunogen in pathogenic treponemes other than T. pallidum subspecies pallidum was also observed (43). Anti-47 K immunogen mAb was nonreactive against rabbit IgG or IgM. mAb directed specifically against the 47 K mol wt immunogen of T. pallidum was examined for strategic functional activities. It was found to be reactive in the microhemagglutination assay for T. pallidum antibodies, the T. pallidum immobilization test, and was found to be capable of significant blockage of attachment of virulent T. pallidum to host cells in tissue culture. Additional significant biological activity for the anti-47 K mol wt immunogen mAb was revealed through results of the in vitro-in vivo neutralization test of Bishop and Miller, in which a 99% or 100% neutralizing activity was demonstrated. The combined data of this study suggest that the 47 K mol wt immunogen of T. pallidum represents an abundant, immunodominant, surface-exposed immunogen possessing potential biological importance in the pathogenesis and immunology of T. pallidum infection. These studies serve to establish the first functionally defined immunogen for T. pallidum, which may represent the major immunogen of the organism.

Plasmid DNA in Treponema pallidum (Nichols): potential for antibiotic resistance by syphilis bacteria.

A plasmid DNA structure (approximate molecular weight = 7.5 X 10(6)) was identified in the human pathogen Treponema pallidum (Nichols). The inability to isolate this plasmid from rabbit host tissue and the total lack of DNA homology of the plasmid with rabbit DNA has confirmed its Treponema pallidum origin. The observation documents a newly recognized and potentially significant genetic capability for Treponema pallidum.

Acquired resistance to Borrelia burgdorferi infection in the rabbit. Comparison between outer surface protein A vaccine- and infection-derived immunity.

Intradermal inoculation of the rabbit with Borrelia burgdorferi, sensu lato, results in the consistent development of erythema migrans (EM), dermal infection, and visceral dissemination of the spirochete. Within 5 mo, EM as well as dermal and visceral infection are cleared and the animals exhibit immunity to reinfection. This study compares infection-derived immunity with acquired resistance resulting from the administration of a lipidated recombinant outer surface protein A (OspA) vaccine presently undergoing human trial. 4 of 11 OspA vaccinated rabbits, challenged intradermally at each of 10 sites with 10(5) low passage B. burgdorferi, developed EM as well as dermal and disseminated infection. After identical challenge, 2 of the 11 infection-immune rabbits developed a dermal infection, but not EM or disseminated infection. Further, ELISA anti-OspA titers did not correlate with the status of immunity for either OspA vaccinated or infection-immune rabbits. Prechallenge ELISA anti-OspA titers were relatively low in the infection-immune group. This study demonstrates that a state of partial immunity to experimental Lyme disease may result that could potentially mask infection. Further, our data strongly suggest that immunogen(s) other than OspA is/are responsible for stimulating acquired resistance in the infection-immune rabbit.

Virulent strain associated outer membrane proteins of Borrelia burgdorferi.

We have isolated and purified outer membrane vesicles (OMV) from Borrelia burgdorferi strain B31 based on methods developed for isolation of Treponema pallidum OMV. Purified OMV exhibited distinct porin activities with conductances of 0.6 and 12.6 nano-Siemen and had no detectable beta-NADH oxidase activity indicating their outer membrane origin and their lack of inner membrane contamination, respectively. Hydrophobic proteins were identified by phase partitioning with Triton X-114. Most of these hydrophobic membrane proteins were not acylated, suggesting that they are outer membrane-spanning proteins. Identification of palmitate-labeled lipoproteins revealed that several were enriched in the OMV, several were enriched in the protoplasmic cylinder inner membrane fraction, and others were found exclusively associated with the inner membrane. The protein composition of OMV changed significantly with successive in vitro cultivation of strain B31. Using antiserum with specificity for virulent strain B31, we identified OMV antigens on the surface of the spirochete and identified proteins whose presence in OMV could be correlated with virulence and protective immunity in the rabbit Lyme disease model. These virulent strain associated outer membrane-spanning proteins may provide new insight into the pathogenesis of Lyme disease.

Rabbit model of Lyme borreliosis: erythema migrans, infection-derived immunity, and identification of Borrelia burgdorferi proteins associated with virulence and protective immunity.

Erythema migrans (EM), persistent skin infection, and visceral dissemination can be induced reproducibly in the adult male New Zealand White rabbit by intradermal injection of as few as 10(3) Borrelia burgdorferi. EM was found to persist for 7 +/- 3 d. Skin culture positivity (infection) cleared within a mean of 6.7 +/- 1.4 wk after infection and similarly visceral infection was not demonstrated after 8 wk; infection-derived immunity to intradermal challenge was evident 5 mo after initial infection. The extent of the protection against EM and dermal infection induced by untreated infection was directly related to the extent of prior in vitro passage of the B31 strain. Initial infection with as few as 4 x 10(3) B31 passage 4 induced complete protection against EM and skin infection upon subsequent challenge with 4 x 10(7) B31, passage 4. Initial infection with B31 passage 27 led to partial protection against EM along with complete protection against skin infection. Initial infection with passage 47 led to partial protection against EM, but conferred no protection against skin infection. Using serum from rabbits fully immune to reinfection, we defined a set of B. burgdorferi proteins present in virulent B31, but absent in the avirulent American Type Culture Collection B31 strain, termed "va" for virulent strain associated. The va proteins of B31 passages 1, 27, and 47 differed strikingly, thus raising the possibility that these changes may relate in a causal way to the differences in induction of protective immunity observed.

The luminescence properties of concanavalin A.

1. The luminescence properties of native concanavalin A, both at room temperature and at 77 degrees K, are similar to those of other proteins containing tyrosine and tryptophan. 2. Binding of methyl alpha-D-glucopyranoside to concanavalin A causes a slight reduction of its fluorescence at room temperature. 3. Removal of Mn2+ and Ca2+ ions from concanavalin A causes a small increase in its fluoresence. The fluorescence: phosphorescence ratio and phosphorescence lifetime of apo-concanavalin A are similar to those of tryptophan. 4. Denaturation of concanavalin A by urea and by guanidine hydrochloride apparently takes place in two stages. Apo-concanavalin A is more easily denatured than the native molecule, but concavalin A combined with methyl alpha-D-glucopyranoside is more resistant to denaturation. 5. The luminescence properties of concanavalin A are pH-dependent. 6. The results have been interpreted in terms of the known structure and properties of concanavalin A.

The low-temperature luminescence properties of bovine alpha-lactalbumin.

The luminescence of bovine alpha-lactalbumin at 77 K has been studied and compared with that of lysozyme. Alpha-Lactalbumin has several unusual properties, including a fluorescence spectrum showing vibrational fine structure, an abnormal phosphorescence spectrum, a high fluorescence: phosphorescence ratio and an abnormal phosphorescence decay. These properties are largely due to the proximity of tryptophan residues to disulphide bonds. Reduction of all these bonds causes considerable changes in alpha-lactalbumin luminescence, as does denaturation in acid solution. Reduction of a single labile disulphide bond has little effect, and the properties of alpha-lactalbumin III, a variant lacking one disulphide bond and one trypotophan residue, are similar to those of the normal protein. Several differences between alpha-lactalbumin and lysozyme are reported. The results support the suggestion that the two tryptophan residues found in the active site cleft of alpha-lactalbumin may be largely responsible for its luminescence.

Factors affecting the luminescence of tryptophan at 77 K.

The effects of solvent polarity, heavy atoms and sulphur-containing compounds on the fluorescence and phosphorescence of indole and tryptophan have been investigated. The low-temperature luminescence of a group of dipeptides containing tryptophan is also reported. The phosphorescence to fluorescence quantum yield ratio and the phosphorescence lifetime are shown to be particularly sensitive to environmental factors. The relevance of these results to the interpretation of protein luminescence spectra is briefly discussed.

Host response to Treponema pallidum in intradermally-infected rabbits: evidence for persistence of infection at local and distant sites.

Intradermal infection of rabbits with Treponema pallidum imitiates rapid and active cellular response at the site of injection. During the first 2 weeks following infection, there is a marked increase in the numbers of organisms at the site of infection. Systemic dissemination of treponemes occurs during the early stage of infection, presumably before the immune response is fully mobilized. The mononuclear infiltration, which is apparent at the lesion site one week postinfection, becomes more pronounced at 2 weeks. The infiltrating cells are predominantly T lymphocytes and macrophages. By 4 weeks postinfection, most of the organisms have been cleared from the primary site; however, low numbers of treponemes survive locally and in distant tissues. Thus, whereas infection with T. pallidum appears to activate immune mechanisms which are capable of clearing most of the organisms from the primary lesion, some organisms are able to evade these mechanisms and persist in vivo.

Monaghan 225 ventilator use under hyperbaric conditions.

The Monaghan 225 ventilator was tested to ambient pressures of 6 atmospheres absolute (ATA) in a hyperbaric chamber. The ventilator would function with delivered tidal volume which was independent of ambient pressure. Ventilatory rate declined in an exponential fashion. At 6 ATA, the ventilatory rate was 45 percent of the preset rate at 1 ATA. By decreasing the circuit resistance and increasing the inspiratory flow rate, the 6 ATA rate could be increased to 72 percent of the 1 ATA value. The maximum minute ventilation of the ventilator at 1 ATA was approximately 48 L/min; at 6 ATA, its maximum was 18 L/min. Synchronized intermittent mandatory ventilation, assist/control, and PEEP functions were satisfactory at 6 ATA. While using 100 percent O2 to power the ventilator at 2.82 ATA, the oxygen leakage was 57.7 L/min (converted to 1 ATA pressure, 20 degrees C), of which 33.7 L/min was successfully scavenged using simple techniques. A minor modification was made to the ventilator, allowing it to be driven by compressed air while maintaining complete flexibility in setting the FIo2. The ventilator has proven stable and reliable in clinical use at ambient pressures up to 6 ATA.

The external validity of Asperger disorder: lack of evidence from the domain of neuropsychology.

The present study compared individuals with high-functioning autism (HFA) and Asperger disorder (AD) in intellectual, motor, visuospatial, and executive function domains. Participants with AD demonstrated significantly higher Verbal and Full Scale IQ scores, significantly larger Verbal-Performance IQ discrepancies, and significantly better visual-perceptual skills than those with HFA. Once the superior intellectual abilities of the AD group were controlled (both statistically through analysis of covariance and by examining IQ-matched subgroups of HFA and AD participants), no significant group differences in motor, visuospatial, or executive functions were evident, save a marginally significant trend toward poorer fine motor performance in the AD group. This suggests that AD may simply be "high-IQ autism" and that separate names for the disorders may not be warranted. The relation of these findings to theories of autism and AD are discussed.

Simultaneous mass spectrometry and thermoconductivity measurements of end-tidal xenon concentrations: a comparison.

Studies in which the rate of local cerebral blood flow is determined by xenon-enhanced computerized tomography require measurement of end-tidal gas. A mass spectrometer and a thermoconductivity detector were compared in measuring the concentration of xenon in end-tidal gas both in humans and in a clinically simulated ("breathing bag") system. The experiments showed that the two instruments provided virtually identical results up to a rate of 18 breaths/min, at which rate recorded concentrations showed marginal degradation with the thermoconductivity detector. Given the absence of a significant difference in the ability of the mass spectrometer and the thermoconductivity detector to measure end-tidal xenon concentration, the thermoconductivity detector appears to be a satisfactory alternative in studies measuring the rate of local cerebral blood flow by enhanced computerized tomography.

Teaching theory of mind: a new approach to social skills training for individuals with autism.

This study examined the effectiveness of a social skills training program for normal-IQ adolescents with autism. Five boys participated in the 4 1/2-month treatment condition; four boys matched on age, IQ, and severity of autism constituted the no-treatment control group. In addition to teaching specific interactional and conversational skills, the training program provided explicit and systematic instruction in the underlying social-cognitive principles necessary to infer the mental states of others (i.e., theory of mind). Pre- and post-intervention assessment demonstrated meaningful change in the treatment group's performance on several false belief tasks, but no improvement in the control sample. No changes, however, were demonstrated on general parent and teacher ratings of social competence for either group.

Autism and autistic behavior in Joubert syndrome.

To determine whether individuals with Joubert syndrome exhibit features of autism as defined by the Diagnostic and Statistical Manual of Mental Disorders-IV (DSM-IV), we examined 11 children with Joubert syndrome using the Autism Diagnostic Interview-Revised and the Autism Diagnostic Observation Schedule-Generic. Three children met DSM-IV criteria for autistic disorder and one for pervasive developmental disorder not otherwise specified. The other seven all demonstrated at least one DSM-IV symptom of autism, but did not meet criteria for a pervasive developmental disorder. Both total number of DSM-IV symptoms and number of social symptoms distinguished the autism and nonautism subgroups. In contrast, the two subgroups displayed similar levels of communication impairments and repetitive or stereotyped behavior. The key to diagnosing autism in Joubert syndrome is to focus on social behaviors, particularly milestones typically achieved very early in life (eg, attending to human voices, showing objects of interest, enjoyment of social interactions). Implications for the role of the cerebellum in nonmotor behavior and for clinical management of Joubert syndrome also are discussed.

New fluorigenic reagents and their fluorescent reactions for amino acid measurements.

Naphthalene-2,3-dicarboxaldehyde, 1-phenylnaphthalene-2,3-dicarboxaldehyde and anthracene-2,3-dicarboxaldehyde have been characterized for use as fluorigenic reagents for the determination of primary amines and amino acids at the microgram level. The reaction conditions, spectral properties and stability of the derivatives (isoindoles) have been investigated with standard amino acids. The results have been compared with other fluorigenic reagents such as ortho-phthalaldehyde.

Phosphorescence of adsorbed molecules at room temperature-the first observations?

The room-temperature phosphorescence of molecules adsorbed on cellulose and other supports seems to have been "discovered" no less than four times.

Determination of pH with acid-base indicators: Implications for optical fibre probes.

Results are presented which clearly illustrate the possibilities and limitations of the use of indicators immobilized on optical fibres, in the determination of pH.

The luminescence properties of some anti-inflammatory and antipyretic drugs.

Twenty anti-inflammatory and antipyretic drugs have been examined for their fluorescence and phosphorescence characteristics, and conditions established for detection of fourteen of them at trace levels.

Luminescence characteristics of several classes of drugs affecting the central nervous system.

The excitation, fluorescence and phosphorescence characteristics of 29 compounds of psychopharmacological interest have been studied in ethanol at 77 k. Phosphorescence lifetimes are reported and the low-temperature luminescence spectra discussed. Luminescence characteristics of several important 1,4-benzodiazepines in acidic, basic and neutral solution are reported.

Photoluminescence and chemiluminescence methods of drug analysis.

The analysis of drugs in biological fluids requires sensitive, selective and convenient methods. Luminescence techniques provide extreme sensitivity and, alone or in conjunction with other methods, excellent selectivity. This paper assesses several recent developments in luminescence analysis, including the use of derivative and synchronous spectroscopy, luminescence immunoassays, room temperature phosphorimetry, and fluorescence excited by chemiluminescence energy transfer.