PubMed HealthSearch

Biomedical subjects

B J Johnson

Publications and source records attributed to B J Johnson.

At least 19 recordsLinked to original sources

The hamster immune response to tick-transmitted Borrelia burgdorferi differs from the response to needle-inoculated, cultured organisms.

The human immune response to natural infection with Borrelia burgdorferi appears to differ from that seen in small mammals infected by needle inoculation. In humans, antibody to outer surface proteins A and B (OspA and OspB) is not detectable until late in infection, but small mammals inoculated with B. burgdorferi produce early antibody to OspA and OspB. To investigate this disparity we compared the immune response in hamsters to B. burgdorferi after needle inoculation with cultured organisms or infected tick homogenates with the immune response after tick transmitted (natural) infection. We determined that the antibody response to OspA and OspB after natural infection of hamsters is similar to that seen in humans, and differs from the antibody response after hamster infection by needle inoculation. High titers of antibody to OspA and OspB were undetectable even 42 wk after bite by B. burgdorferi-infected ticks. The failure to produce antibody to OspA and OspB was not dependent on challenge dose, because animals inoculated by needle with low doses (1 x 10(5) to 1 x 10(6) cells) of B. burgdorferi produced antibody to OspA and OspB. A rapid but limited anti-41-kDa response was observed. One possible new Ag, 43 kDa (p43), was identified. The antibody response to p43 was independent of the route of inoculation. Our results suggest that the hamster immune response to tick-transmitted Borrelia burgdorferi differs from the response to needle inoculated, cultured organisms.

Animals

D-1 and D-2 receptor mediation of sensorimotor behavior in rats depleted of dopamine during development.

The effects of selective D-1 and D-2 antagonists on sensorimotor behavior were studied in rats treated with 6-OHDA or its vehicle solution on either postnatal day 3, 20, or 35. Blockade of either D-1 or D-2 receptors induced akinesia and somatosensory neglect in adults treated with vehicle at any of the three ages. The behavioral effects of antagonists on rats with 6-OHDA-induced dopamine depletions varied as a function of the age at the time of damage. Adults depleted of DA on Day 35 exhibited behavioral deficits after either D-1 or D-2 blockade and at doses that were ineffective in controls. Adults depleted of DA on Day 20 exhibited deficits after either D-1 or D-2 blockade but were not any more sensitive than were controls. In contrast, adults depleted of DA on Day 3 were insensitive to the behavioral effects of D-1 or D-2 blockers but were impaired after the dual administration of both antagonists. Moreover, simultaneous administration of subthreshold doses of D-1 and D-2 antagonists produced behavioral deficits in controls and rats depleted on Day 3. These data demonstrate that activity within residual DA neurons remains critical for the expression of sensorimotor behavior in rats depleted of DA during development. However, the specific contribution of D-1 and D-2 receptors to these behaviors depends upon the animals' age at the time of depletion.

Afferent Pathways

Bovine abortions attributable to Listeria ivanovii: four cases (1988-1990).

During a 3-year period, 4 cases of bovine abortion attributable to Listeria ivanovii were diagnosed from 243 bovine fetuses submitted for diagnostic evaluation. Listeria monocytogenes was isolated only once from a bovine fetus during this same time period. Pathologic findings were similar to those seen in abortions attributable to L monocytogenes. Consistent management factors were not recognized and breed susceptibility was not apparent. Listeria ivanovii is most often associated with abortions from sheep and is rarely reported from cattle. On the basis of findings in this study, L ivanovii must be included as a potential cause of bovine abortions.

Abortion, Veterinary

An association between complete and incomplete stress fractures of the humerus in racehorses.

Twenty-one horses had a complete unilateral humeral fracture during race training or racing at a California racetrack during the period 24 February 1990 to 10 July 1991. Fractures occurred approximately equally in left and right limbs, and in males and females. Most fractures occurred during training, and in 2- and 3-year-old horses. Only 5 of 16 Thoroughbred horses with known racing records had previously raced more than once, and their mean time between races was less than the time between their last race and fracture (P = 0.07). Ten of 13 humeri studied further had gross evidence of periosteal callus bridging one portion of the fracture line, indicative of a pre-existing stress fracture.

Animals

Detection of Borrelia burgdorferi in ticks by species-specific amplification of the flagellin gene.

We developed a polymerase chain reaction (PCR) that specifically amplifies a fragment of the flagellin gene (fla) of Borrelia burgdorferi, the causative agent of Lyme disease. This fla target, amplified with nested primers, was conserved among all 80 strains of B. burgdorferi tested. Strains examined included cultures from ticks, humans, and rodents from major B. burgdorferi-endemic regions of the United States and parts of Europe and Asia. Templates from B. hermsii, B. parkeri, B. turicatae, and B. coriaceae were not amplified, nor were eukaryotic DNAs from three tick genera. Several host DNAs potentially present in a tick blood meal also were not amplified. Approximately six B. burgdorferi per PCR reaction could be detected by ethidium bromide staining of amplified DNA. Colony-raised Ixodes dammini were used to evaluate the method. One infected nymph in a pool of 40 ticks was routinely detected. The specificity of the assay for detecting B. burgdorferi-infected ticks in pools was 94% (29 of 31). This protocol should prove useful for assessing infection rates in other putative arthropod vectors.

Animals

Variants of Venezuelan equine encephalitis virus that resist neutralization define a domain of the E2 glycoprotein.

Stable neutralization (N) escape variants of Venezuelan equine encephalitis (VEE) virus were selected by anti-E2 glycoprotein monoclonal antibodies (MAbs) that neutralize viral infectivity, block viral hemagglutination, and passively protect mice. The nucleotide sequence of the E1, E2, and E3 genes of four variants revealed a clustering of single mutations in a domain spanning E2-182 to E2-207. The conformation of this short linear sequence affects antigenicity in the N domain because reduction and alkylation of virus disrupted binding of some E2 neutralizing MAbs. Serologic evidence for interaction of E2 epitopes also was obtained. Mutations in the N domain of VEE virus did not alter the kinetics of binding to Vero cells. They did, in some cases, produce attenuation of virulence in mice.

Amino Acid Sequence

Utilization of a radioiodinated bile salt for kinetic studies and hepatic scintigraphy. Studies in nonhuman mammals.

An 125I- or 131I-labeled bile salt derivative, cholylglycyliodohistamine, has been synthesized and purified. The bile salt derivative is rapidly cleared from the circulation when injected intravenously into rats and rabbits. Ten minutes after injection, approximately 50% of the recovered bile salt derivative was in the jejunum and ileum, and 36% was found in the liver. Sixty minutes after injection, 99% of the recovered radioactivity was found in the luminal gastrointestinal tract. The isotope was cleared from the circulation of rabbits with a t1/2 of approximately 2 min. Hepatic scintigraphy using rabbits demonstrated rapid uptake by the liver and excretion into the intestine. Quantitative analysis of scintigraphy showed an uptake rate of 14%/min and a subsequent excretory rate of 4.6/min.

Animals

Mobilization of colonic kallikrein following pelvic nerve stimulation in the atropinized cat.

1. Pelvic nerve stimulation (p.n.s.) in cats induces atropine-resistant colonic vasodilatation and colonic contraction. The effects of this on cat colon are mimicked by synthetic bradykinin infusions. The present study examines the effect of p.n.s. on the activation of kallikrein, the kinin-forming enzyme present in colonic tissue and its effects on the plasma kinin system in the atropinized cat.2. Mean level (+/- S.D.) of mucosal kallikrein was found to be about 37 times higher in unstimulated colonic mucosa (300 +/- 100 ng bradykinin equivalents min(-1)g(-1)) than in the underlying muscle (8.2 +/- 6.3 ng bradykinin equiv min(-1)g(-1)).3. After a p.n.s. of 5 min, mean kallikrein level in colonic muscle was 7.3 +/- 3.5 ng bradykinin equiv min(-1)g(-1), which was not significantly different from the control muscle kallikrein. However, there was an 86% fall in mucosal kallikrein to 41.3 +/- 34.7 ng bradykinin equiv min(-1)g(-1) after 5 min p.n.s., indicating a rapid activation and secretion of mucosal kallikrein.4. Secretion of mucosal kallikrein was paralleled by specific depletion of plasma kininogen, the precursor of active kinin in blood draining the colon. The mean plasma kininogen level fell to 79 and 68% of the prestimulated value (3.1 +/- 1.1 S.D. mug bradykinin equiv per ml. plasma) after 5 and 10 min p.n.s. respectively. Total plasma protein and haematocrit remained unaltered excluding non-specific changes due to protein extravasation or haemodilution and indicating utilization of the plasma kinin precursor.5. Following 2 hr p.n.s., raised levels of kallikrein were detected in both colonic muscle (28 +/- 2.0 bradykinin equiv min(-1)g(-1)) and mucosa 434 +/- 118 ng bradykinin equiv min(-1)g(-1)). Preliminary studies using a kallikrein inhibitor indicated that the increased kallikrein levels originated from plasma.6. Direct stimulation of the parasympathetic pelvic nerve in the atropinized cat thus produced activation of the plasma kinin system in the colon and formation of free kinins may be responsible for the mucosal vasodilatation and strong motor contraction which is not blocked by large doses of atropine. The observation that prolonged stimulation causes extravasation of plasma kallikrein, a potential inflammatory mediator, into the tissues may be of clinical significance.

Animals

Complement fixing site of human IgG1.

A number of chemical modifications were made to purified human IgG1. The effects of these modifications on the complement activity of the immunoglobulin were studied using aggregation on latex and measuring the consumption of the complement by CH50-quantitation. Tryptophan and tyrosine are implicated in the complement fixing site of this human immunoglobulin, and an arginine moiety probably provides a binding site for the complement.

Adsorption

Lipid-protein interactions: detergent binding to L-glutamic acid dehydrogenase.

A nonionic detergent was found to bind to the enzyme L-glutamic acid dehydrogenase [L-glutamate:nicotinamide adenine dinucleotide phosphate oxidoreductase (deaminating) EC 1.4.1.3]. The amount bound was 17 moles of detergent/mole of enzyme, which, however, was not sufficient for the enzyme to be included in a detergent micelle.

Chromatography, Gel

Interrelation between two anticomplement cobra venom factors isolated from crude Naja naja cobra venom.

Two moieties occurring in crude Naja naja cobra venom were found to possess anticomplement activity. Both materials possessed similar molecular weights and specific activities but dissimilar elution profiles upon ion-exchange chromatography. The anticomplement activities of these materials were maintained upon digestion with neuraminidase, and their elution profiles from cation-exchange chromatography became identical after this treatment. It was concluded that the differences between the two anticomplement materials were due to their different sialic acid contents.

Animals

An 125I radioimmunoassay for primary conjugated bile salts.

Cholylglycylhistamine, a derivative of cholic acid, has been synthesized and characterized. This derivative has been iodinated using Na125I and chloramine-T and purified free from unlabeled cholylglycylhistamine. Application of this iodinated bile salt derivative to radioimmunoassay of bile salts in human serum is reported. Antibody titers have uniformly increased over titers used in tritium-based assays; some antibodies are usable in dilutions of 1:80,000. The radioimmunoassay described here was found to measure predominantly the primary conjugated bile salts. Sensitivity has been maintained, with the least detectable amount being 0.5 pmoles per assay tube. Normal values in human serum are 3.47 +/- 2.16 (SD) nmoles per ml.

Animals

Lipid-protein interactions: enhancement of enzyme activity of L-glutamic acid dehydrogenase by nonionic detergents.

Five nonionic detergents enhanced the activity of L-glutamic acid dehydrogenase [L-glutamate:nicotinamide adenine dinucleotide phosphate oxidoreductase (deaminating) (EC 1.4.1.3)]. These detergents activated the enzyme toward alpha-ketoglutaric acid reduction, causing a decrease in the sensitivity of the enzyme to allosteric regulation by guanosine 5-triphosphate. There was also a diminution of the enhancing effect of the modifier adenosine 5-diphosphate on the enzyme's L-glutamic acid dehydrogenase activity. These detergents may cause a conformational change in the enzyme, and this change could lead to an increase in the binding of the substrates for the alpha-ketoglutaric acid reduction. Accompanied with this conformational change would be a decrease in the binding of the modifier guanosine 5'-triphosphate, with no concomitant change in the binding of the adenosine 5'-diphosphate modifier.

Adenosine Diphosphate

Investigations of the complement-fixing sites of immunoglobulins.

Indirect evidence for the complement-fixing site of human IgG1 has been found by using chemical modification of the immunoglobulin. These results implicate the amino acid residues, tryptophan, tyrosine, and arginine. Furthermore, by the use of synthetic peptides it has been shown that the area surrounding the tryptophan and tyrosine at positions 277 and 278 probably constitutes the locus of the complement-fixing site of human IgG1. By analogy, the tryptophan at position 488 of human IgM is implicated in the complement-fixing site of this immunoglobulin. Evidence for this is presented by a peptide which mimics the sequence 487-491 of human IgM and possesses the ability to consume complement. The relative complement-fixing ability of this peptide is 200 times less effective than human IgG1.

Amino Acid Sequence