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Biomedical subjects

A A Jacobs

Publications and source records attributed to A A Jacobs.

At least 19 recordsLinked to original sources

Safety and efficacy of an oil-adjuvant vaccine against haemorrhagic septicaemia in buffalo calves: cross-protection between the serotypes B:2,5 and E:2,5.

The safety, efficacy and duration of immunity of an improved oil-adjuvant vaccine against haemorrhagic septicaemia, containing inactivated cells of Pasteurella multocida serotype B:2,5, were tested in young buffalo calves in Pakistan. For safety testing, five buffalo calves were vaccinated intramuscularly with twice the normal dose, and six weeks later with a normal dose. Except for a transient rise in rectal temperature at six hours after the vaccinations, no systemic reactions were observed. The buffaloes remained in good condition and had a normal appetite. No local reactions were observed at the injection site. For efficacy testing two trials were carried out. In the first, buffalo calves were vaccinated intramuscularly either with two doses two-and-a-half months apart, or with a single dose, or left unvaccinated. They were challenged subcutaneously with virulent P multocida after eight, 13 or 15 months. After challenge at eight months the four buffaloes given two doses and the buffalo given one dose were protected, whereas the control animal developed the typical signs of the disease. After the challenges at 13 and 15 months, the vaccinated animals were still protected whereas the control animals died. In the second trial, buffalo calves were vaccinated intramuscularly either with two doses two months apart, or with a single dose at two months or left unvaccinated. The buffaloes were challenged after eight or 14 months. After challenge at eight months the four control animals died, whereas three of the four buffaloes given a single dose were protected. After challenge at 14 months, the three control animals died, whereas four of the five buffaloes given two doses and both the buffaloes given a single dose were protected. To test for cross-protection against the heterologous serotypes E:2,5 and B:3,4, groups of mice were vaccinated once or left unvaccinated. Four weeks later, the vaccinated and control groups were challenged with a dilution series of the different challenge cultures. The vaccine appeared to induce protection against challenge with different strains of serotypes B:2,5 and E:2,5 but not against strains of serotype B:3,4.

Adjuvants, Immunologic↗

Investigations towards an efficacious and safe strangles vaccine: submucosal vaccination with a live attenuated Streptococcus equi.

As part of a search for a safe and efficacious strangles vaccine, several different vaccines and different vaccination routes were tested in foals. The degree of protection was evaluated after an intranasal challenge with virulent Streptococcus equi by clinical, postmortem and bacteriological examinations. Inactivated vaccines containing either native purified M-protein (500 microg per dose) or whole S equi cells (10(10) cells per dose) administered at least twice intramuscularly at intervals of four weeks, did not protect against challenge. Different live attenuated S equi mutants administered at least twice at intervals of four weeks by the intranasal route were either safe but not protective or caused strangles. In contrast, a live attenuated deletion mutant administered intramuscularly, induced complete protection but also induced unacceptable local reactions at the site of vaccination. Submucosal vaccination in the inner side of the upper lip with the live attenuated mutant at > or =10(8) colony-forming units per dose, appeared to be safe and efficacious in foals as young as four months of age. The submucosal vaccinations caused small transient swellings that resolved completely within two weeks, and postmortem no vaccine remnants or other abnormalities were found at the site of vaccination.

Animals↗

DNA rearrangements in the flagellin locus of an flaA mutant of Campylobacter jejuni during colonization of chicken ceca.

Campylobacter jejuni is an enteropathogen for humans but commensal for chickens. In both hosts, the flagella and motility are important colonization factors. The flagellin gene is duplicated in Campylobacter, but only one flagellin gene, flaA, is sufficient for motility. We found that, during colonization of the chicken intestine, a nonmotile flaA mutant of C. jejuni underwent rearrangements within its flagellin locus, thereby regaining its motility and colonization capacity. In contrast, in vitro motile revertants isolated from liquid culture showed different flagellin DNA rearrangements than after reversion in the chicken.

Animals↗

Efficacy of a commercially available coryza vaccine against challenge with recent South African NAD-independent isolates of Haemophilus paragallinarum in chickens.

In South Africa the incidence of NAD-independent Haemnophilus paragallinarum isolation from clinical cases is increasing. This study was carried out to test whether a commercially available coryza vaccine (Nobilis Coryza, Intervet International BV) could protect chickens against challenge with recent NAD-independent isolates. SPF chickens were vaccinated twice at 3 and 7 weeks of age and were challenged at 9 weeks of age with 5 different NAD-independent isolates of serotype A or C-3. The results after challenge show that the coryza vaccine induces good protection against challenge with the different South African NAD-independent isolates of H. paragallinarum, including serotype C-3.

Animals↗

The conserved undecapeptide shared by thiol-activated cytolysins is involved in membrane binding.

Thiol-activated cytolysins share a conserved hydrophobic, Trp-rich undecapeptide that is suggested to be involved in membrane binding and intercalation. The neutralizing monoclonal antibody PLY-5 recognizes all members of this toxin family and peptide mapping assigned its epitope to the undecapeptide motif. This antibody inhibited binding of the toxins to host cell membranes and the epitope was no longer available for binding when a preformed toxin/membrane complex was tested. These results confirm the model of cytolysin binding suggested by structural data.

Amino Acid Sequence↗

Characterisation of the gene encoding suilysin from Streptococcus suis and expression in field strains.

The gene encoding suilysin was cloned from Streptococcus suis serotype 2 strain P1/7. Analysis of the nucleotide and translated amino acid sequence confirmed suilysin to be a member of the thiol activated cytolysin group (TACY). The pneumolysin from Streptococcus pneumoniae is the most closely related orthologous gene known. Suilysin was overexpressed in E. coli in an active haemolytic form. A strong correlation between the presence of the sly gene and haemolytic activity in the supernatant of S. suis field strains was found. Of 158 strains tested, 63% contained the gene. Within the (most prevalent) serotype 2, the sly gene was demonstrated in 95% of the strains isolated in Eurasia, but only in 7% of the strains from North America.

Animals↗

Continuous venovenous hemofiltration and hemodialysis after orthotopic heart transplantation.

Orthotopic heart transplantation (OHT) is an accepted treatment for heart failure refractory to medical management. International registry data reported a 1-year survival after OHT of 79%. The risk factors associated with requiring continuous venovenous hemofiltration or hemodialysis and mortality in patients requiring continuous venovenous hemofiltration (CVVH)/hemodialysis (HD) have not been studied, since improvements have been made in OHT. We tested the hypothesis that requiring CVVH/HD in the immediate posttransplantation period increases mortality after OHT. We studied pretransplantation factors to predict those patients who would need CVVH/HD. Patients undergoing OHT from 1995 to 1996 were studied. Fifty-two patients underwent OHT. Eight patients (15%) needed CVVH/HD. Initial immunosuppression included cyclosporine, azathioprine, and solumedrol. Patients were changed to antilymphocyte therapy if they had oliguric acute renal failure, T or B cell incompatibility, or biopsy-proven rejection. Overall, survival at 1 year was 84.6%. Survival in patients not needing CVVH/HD was 91%, and survival in patients needing CVVH/HD was 36.5%. For each year of age over 55 years, patients had a relative risk of requiring CVVH/HD of 7.

Acute Kidney Injury↗

Protection of experimentally infected pigs by suilysin, the thiol-activated haemolysin of Streptococcus suis.

Three groups of three pigs were vaccinated either with vaccine VAC-SLY, containing purified suilysin derived from Streptococcus suis strain P1/7 (serotype 2), or with vaccine VAC-SCF, containing most of the other extracellular antigens produced by strain P1/7 (but essentially free from suilysin), or with a placebo vaccine. The pigs were vaccinated twice at four weeks and six weeks of age and were challenged intravenously with S suis strain P1/7 at eight weeks of age. On the day of challenge, only the VAC-SLY vaccinated pigs showed an increase in haemolysin neutralisation titre. After challenge the placebo vaccinated pigs developed severe clinical signs characterised by lameness involving several joints, a depressed appearance, high temperatures and/or neurological signs. The VAC-SCF vaccinated pigs showed the same clinical signs but less severely. The VAC-SLY vaccinated pigs were the least affected and showed only mild signs which subsided more quickly than those of the other groups. A post mortem investigation and histology of brain tissue samples confirmed the clinical findings; fibrinous arthritis was less severe and less frequently observed in the VAC-SLY vaccinated pigs than in the VAC-SCF or placebo vaccinated pigs, and none of the VAC-SLY vaccinated pigs had meningitis whereas two of the VAC-SCF and two of the placebo vaccinated pigs did so. All the samples of brain, lung and tarsus taken from the VAC-SLY vaccinated pigs were sterile whereas S suis was reisolated from most of these tissues from the other groups.

Animals↗

TNF-alpha stimulates increased plasma membrane guanine nucleotide binding protein activity in polymorphonuclear leukocytes.

TNF-alpha enhances the response of polymorphonuclear leukocytes (PMN) to chemoattractants: however, the mechanism by which this occurs is unclear. We addressed the hypothesis that TNF-alpha enhances the PMN response to chemoattractants by increasing chemoattractant receptor transmembrane signaling, using fMLP as the model chemoattractant. fMLP-stimulated guanine nucleotide binding (G) protein activation was significantly increased in plasma membranes isolated from PMNs exposed to TNF-alpha 100 U/ml for 10 minutes (TNF-M), compared to membranes from control cells (CM). Formyl peptide receptor number and affinity were not significantly different in CM and TNF-M. Gi and Gs content were increased in TNF-M as measured by pertussis toxin and cholera toxin (CT) catalyzed ADP-ribosylation, respectively. The increased Gi was coupled to the formyl peptide receptor as shown by receptor-specific CT labeling of Gi. Immunoblot analysis showed that both G alpha i2 and G alpha 3 were increased in TNF-M. The functional activity of the increased G protein content was demonstrated by increased NaF-stimulated phospholipase D activity in TNF-alpha-treated PMNs. We conclude that TNF-alpha rapidly stimulates increased PMN plasma membrane expression of G proteins that couple formyl peptide receptors to effector enzymes. Regulation of G protein expression may be a significant mechanism by which TNF regulates PMN function.

Cell Membrane↗

Chemoattractant receptor-specific differences in G protein activation rates regulate effector enzyme and functional responses.

The hypothesis that disparate neutrophil functional responses to various chemoattractants are regulated by receptor-specific rates of G protein activation was examined in HL-60 granulocytes. The initial rates of G protein activation and the affinity of receptor-stimulated G proteins for GTP gamma S in HL-60 membranes stimulated by fMet-Leu-Phe, C5a, and leukotriene B4 (LTB4) differed significantly among the chemoattractants, with a rank order of fMet-Leu-Phe > C5a > LTB4. Equilibrium GTP gamma S binding showed that all three chemoattractants activated a common pool of G proteins. Stimulation of phospholipase D activation, measured as phosphatidylethanol generation, and superoxide release in intact cells also occurred with a rank order of fMet-Leu-Phe > C5a > LTB4. On the other hand, the rank order of receptor affinities for ligand and of the EC50 of chemoattractant stimulation of GTP gamma S binding was C5a > LTB4 > fMet-Leu-Phe. C5a and LTB4 receptor densities were similar but were less than formyl peptide receptor density. Graded pertussis toxin treatment proportionally reduced superoxide release and phospholipase D activation to all three chemoattractants. The results suggest that receptor-specific differences in G protein affinity for guanine nucleotides lead to different rates of guanine nucleotide exchange and, thereby, contribute to disparate effector enzyme and functional responses.

Amino Acid Sequence↗

Tangential migration of neurons in the developing cerebral cortex.

The mammalian cerebral cortex is divided into functionally distinct areas. Although radial patterns of neuronal migration have been thought to be essential for patterning these areas, direct observation of migrating cells in cortical brain slices has revealed that cells follow both radial and nonradial pathways as they travel from their sites of origin in the ventricular zone out to their destinations in the cortical plate (O'Rourke, N.A., Dailey, M.E., Smith, S.J. and McConnell, S.K. (1992) Science 258, 299-302). These findings suggested that neurons may not be confined to radial migratory pathways in vivo. Here, we have examined the patterns of neuronal migration in the intact cortex. Analysis of the orientations of [3H]thymidine-labeled migrating cells suggests that nonradial migration is equally common in brain slices and the intact cortex and that it increases during neurogenesis. Additionally, cells appear to follow nonradial trajectories at all levels of the developing cerebral wall, suggesting that tangential migration may be more prevalent than previously suspected from the imaging studies. Immunostaining with neuron-specific antibodies revealed that many tangentially migrating cells are young neurons. These results suggest that tangential migration in the intact cortex plays a pivotal role in the tangential dispersion of clonally related cells revealed by retroviral lineage studies (Walsh, C. and Cepko, C. L. (1992) Science 255, 434-440). Finally, we examined possible substrata for nonradial migration in dorsal cortical regions where the majority of glia extend radially. Using confocal and electron microscopy, we found that nonradially oriented cells run perpendicular to glial processes and make glancing contacts with them along their leading processes. Thus, if nonradial cells utilize glia as a migratory substratum they must glide across one glial fiber to another. Examination of the relationships between migratory cells and axons revealed axonal contacts with both radial and nonradial cells. These results suggest that nonradial cells use strategies and substrata for migration that differ from those employed by radial cells.

Animals↗

Role of carboxylmethylation in chemoattractant receptor-stimulated G protein activation and functional responses.

The role of G protein gamma subunit carboxylmethylation was examined in HL-60 granulocytes using an inhibitor of S-adenosylmethionine-dependent methylation, periodate-oxidized adenosine (Adox). A 40-60% reduction in gamma subunit carboxyl-methylation was associated with attenuation of fMet-Leu-Phe-stimulated GTP gamma S binding and GTP hydrolysis, while plasma membrane density of formyl peptide receptors, alpha i2, alpha i3, beta, gamma 5, and gamma 7 were not reduced. Reduced pertussis toxin-catalyzed ADP-ribosylation was re-established by in vitro methylation or addition of transducin beta gamma subunits. Superoxide release and inositol phosphate generation stimulated by fMet-Leu-Phe were significantly inhibited by Adox treatment. Carboxylmethylation contributes to transmembrane signalling and functional responses by enhancing association of alpha and beta gamma subunits.

Adenosine↗

Identification, purification, and characterization of a thiol-activated hemolysin (suilysin) of Streptococcus suis.

The present report describes the identification, purification, and characterization of a hemolysin produced by Streptococcus suis type 2. The hemolysin was purified from the culture supernatant by using different filtration steps, Superose-12 column chromatography, and selective (NH4)2SO4 precipitation. The purified hemolysin, designated suilysin, had an apparent molecular mass of 54,000 Da and exhibited a specific activity of 0.7 x 10(6) hemolytic units per mg. Suilysin appeared to belong to a family of toxins known as the thiol-activated toxins, with which it had several characteristics in common: loss of activity upon oxidation, reactivation upon reduction, and inhibition of activity by small amounts of cholesterol. The N-terminal amino acid sequence of suilysin showed many similarities with parts of the deduced N-terminal amino acid sequences of perfringolysin O, streptolysin O, listeriolysin O, alveolysin, and pneumolysin. Mice immunized with a vaccine containing purified suilysin appeared to be completely protected against a lethal S. suis type 2 challenge, indicating that suilysin is an important factor and that the neutralization of this single factor is sufficient to protect mice against the detrimental effects of an S. suis type 2 infection. Most of the different (serotype) strains appeared to secrete hemolytic activity which was biochemically and immunologically indistinguishable from suilysin into the culture supernatant in vitro, indicating that suilysin might be a cross-protection factor.

Amino Acid Sequence↗

Feline bordetellosis: challenge and vaccine studies.

Four eight-week-old cats, shown to be free from feline calicivirus, feline herpesvirus and Chlamydia psittaci were challenged with an aerosol of Bordetella bronchiseptica. Within five days the cats developed signs of respiratory disease, characterised by nasal discharge, sneezing, spontaneous or induced coughing and dry or wet rales at auscultation. These signs were present for about 10 days, after which they began to resolve. To test the protective capacity of an experimental fimbrial antigen-based subunit vaccine, 10 kittens were vaccinated twice, with two weeks between the vaccinations, and five kittens were left unvaccinated. Two weeks after the booster the 15 kittens were challenged with an aerosol of B bronchiseptica as the sole pathogen. On the day of challenge the vaccinated kittens had a mean bordetella antibody titre of 2(9.5) whereas the control cats remained seronegative (titre < 2(2)). The control cats developed signs of respiratory disease after challenge, whereas the vaccinated cats were almost completely protected. The degrees of protection against rhinitis, sneezing, spontaneous or induced coughing, and dry or wet rales at auscultation were 100 per cent, 95 per cent, 95 per cent and 100 per cent, respectively. Furthermore, the vaccinated kittens cleared the challenge bacteria more quickly than the controls, resulting in a reduction of 80 per cent on days 15 and 18 after challenge and a reduction of 99 per cent on days 22 and 29 after challenge. The results show that B bronchiseptica can act as a primary pathogen in cats and that a vaccine containing the fimbrial antigen induces a protective immune response.

Animals↗

Rapid degradation of NAD by retinoic acid-differentiated HL-60 granulocyte membranes prevents ADP ribosylation.

Pertussis and cholera toxins failed to ADP-ribosylate G alpha s and G alpha i in membranes isolated from retinoic acid-differentiated HL-60 granulocytes, although G alpha i subunits were present. NAD was rapidly degraded in the presence of these membranes, primarily to ADP-ribose, to less than 10% of initial activity by 5 min. Metabolism of NAD was heat labile and inhibited by NADP, but not by imidazole or isonicotinic acid hydrazine. Pertussis toxin uncoupled LTB4 receptors from G proteins in intact retinoic acid-differentiated HL-60 cells. Retinoic acid differentiation stimulates expression of a unique NAD-glycohydrolase activity in HL-60 granulocytes which prevents ADP-ribosylation in isolated membranes, but not intact cells.

Adenosine Diphosphate Ribose↗

Desensitization by protein kinase C activation differentially uncouples formyl peptide receptors from effector enzymes in HL-60 granulocytes.

The hypothesis that protein kinase C (PKC) participates in agonist-mediated desensitization of formyl peptide receptors in HL-60 granulocytes was tested. fMet-Leu-Phe and leukotriene B4(LTB4) produced homologous desensitization of agonist-stimulated intracellular calcium transients. Pre-treatment with the PKC activator, phorbol myristate acetate (PMA; 10 nM), abolished both fMet-Leu-Phe and LTB4-stimulated calcium transients. Membranes prepared from control HL-60 granulocytes (NM) or cells treated with 10 nM PMA (PMA-M) demonstrated increased formyl peptide receptor and G protein density, as determined by radioligand binding and pertussis toxin- and cholera toxin-catalysed ADP ribosylation. fMet-Leu-Phe stimulation of guanosine 5'-[gamma-thio]-triphosphate (GTP gamma S) binding and GTP hydrolysis and GDP inhibition of fMet-Leu-Phe binding were not different between NM and PMA-M. Pre-treatment with 10 nM PMA did not inhibit subsequent fMet-Leu-Phe-stimulated superoxide generation or phospholipase D activation. We conclude that PKC desensitizes fMet-Leu-Phe-stimulated phospholipase C, but not phospholipase D, responses and that PKC activation does not mediate agonist-induced desensitization of formyl peptide receptors.

Autoradiography↗

Developmental profile of polyadenylated and non-polyadenylated GABAA receptor subunit mRNAs.

The ratio of mRNA not selected for polyadenylation (non-poly(A)+ selected) to mRNA selected for polyadenylation (poly(A)+) for the beta 1, alpha 1 and gamma 2 subunits of the GABAA receptor complex was examined in rats as a function of age. RNA was extracted from whole brain of rats that were either 0, 1, 3, 5 or over 60 days of postnatal age. Poly(A)+ mRNA was purified by oligo(dT)-cellulose chromatography. Non-poly(A)+ selected mRNA and poly(A)+ mRNA for the GABAA receptor beta 1, alpha 1 and gamma 2 subunits were examined by Northern blot analysis using cDNA probes specific for these subunits. Levels of GABAA receptor beta 1 subunit mRNA were also examined by solution hybridization analysis with a beta 1 riboprobe. Analysis of Northern blots revealed that levels of poly(A)+ beta 1 subunit mRNA were highest at 0 days of age, but decreased and reached adult levels by 5 days of postnatal age. However, levels of the beta 1 subunit message extracted from non-poly(A)+ selected mRNA were not significantly different at any of the ages examined, suggesting the existence of a population of beta 1 subunit mRNA that is not polyadenylated. The age-related discrepancy between beta 1 subunit levels measured in non-poly(A)+ selected mRNA and poly(A)+ mRNA was also observed using solution hybridization analysis. In contrast, levels of both non-poly(A)+ selected mRNA and poly(A)+ mRNA for the alpha 1 subunit of the GABAA complex increased from 0 days of age to adulthood. Similarly, levels of both non-poly(A)+ selected mRNA and poly(A)+ mRNA for the GABAA receptor gamma 2 subunit increased with age.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗