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Biomedical subjects

R B Johnson

Publications and source records attributed to R B Johnson.

At least 109 records · Page 6Linked to original sources

Preparation of bone for high-voltage electron microscopic radioautography.

This paper presents a reliable technique for the preparation of radioautographic specimens for study by high-voltage electron microscopy. Tissues are fixed by ventricular perfusion of aldehydes, sectioned, collected on slides, and coated with nuclear track emulsion using a coating machine. Sections are developed after 17 weeks' exposure in either undiluted Microdol-X or diluted D-19 developer, fixed in 25% sodium thiosulfate, and stained through the emulsion with uranyl acetate in absolute methanol and aqueous lead citrate. The technique produces sections of even thickness and staining density with negligible numbers of background grains and non-specific labeling. In this report, the efficacy of this technique for the study of the periosteal surface of demineralized alveolar bone specimens is demonstrated.

Animals↗

Pasteurella multocida infections in mice with reference to haemorrhagic septicaemia in cattle and buffalo.

Haemorrhagic septicaemia (HS) is an infectious disease of cattle and buffalo caused by particular serotypes of Pasteurella multocida and is one of the most economically important livestock diseases in South-East Asia. While HS has been recognized for many years, very little is understood about the disease, primarily because of the expense of cattle and a lack of suitable animal models. The suitability of using mice to study HS was assessed using parameters such as the critical pathogenic dose, kinetics of infection, pathology of disease and resistance to reinfection. Pasteurella multocida M1404, the type strain for Carter group B, the serotype responsible for Asian HS, was injected intraperitoneally into BALB/c mice. As few as 20 colony forming units produced an overwhelming septicaemia in mice in less than 30 h. The kinetics of infection demonstrated a very rapid in vivo multiplication rate. There was no evidence of inhibition of bacterial cell growth by natural host defence mechanisms, even with the very small inocula used. The gross pathology of the disease in mice was characterized by splenomegaly, lymphadenopathy and petechial haemorrhages similar to that observed in cattle and buffalo with HS. Mice were found to develop a short-lived resistance to reinfection following a primary infection which had been successfully treated with antibiotics. The mouse would seem to provide an ideal tool by which to study HS, but warrant further studies in order to be able to critically assess it as a model for this economically important disease.

Animals↗

Azithromycin in the treatment of sexually transmitted disease.

One hundred and eighty-two patients were enrolled in a randomized third-party blinded study to assess the efficacy and safety of azithromycin in the treatment of sexually transmitted diseases. Three regimens of azithromycin, including a single oral dose, were compared with a standard treatment with doxycycline. The patients were followed for four weeks. Efficacy was evaluated in 168 patients (113 azithromycin, 55 doxycycline). Fourteen patients had negative cultures or did not come for all follow-up visits. Of the 168, 138 were infected with Chlamydia trachomatis, 43 with Neisseria gonorrhoeae, and 45 with Ureaplasma urealyticum. Ninety-six per cent of patients with chlamydial infections and 92% of those with gonorrhoea were cured with azithromycin. Two patients infected with N. gonorrhoeae, four with C. trachomatis and six with U. urealyticum had positive cultures on follow-up visits after receiving azithromycin. Of these 11 patients with positive cultures on follow-up visits, seven (five with U. urealyticum and two with C. trachomatis) violated the protocol by having intercourse with infected individuals during the study. Azithromycin was very well tolerated; one patient complained of mild abdominal pain shortly after receiving the drug, seven patients complained of mild nausea and two patients had mild diarrhoea.

Adolescent↗

The pharmacokinetics of azithromycin in human serum and tissues.

The pharmacokinetics of azithromycin, a new azalide antibiotic, were examined in man. Approximately 37% of a single oral dose of 500 mg was bioavailable and produced a peak serum concentration of 0.4 mg/l. Multiple dose regimens (two doses of 500 mg separated by 12 h and followed by 500 mg qds for five days, or two doses of 250 mg separated by 12 h and followed by 250 mg qds for nine days) produced only slight increases in peak serum concentrations. The serum protein binding of azithromycin declined from about 50% at 0.02 mg/l to 12% at 0.5 mg/l. Tissue concentrations of azithromycin were much higher than serum concentrations. After two 250 mg doses 12 h apart, peak azithromycin concentrations exceeded 3 mg/kg in prostate, tonsil and many other tissues. Concentrations in tissues declined with apparent half-lives of 2.3 days in prostate and 3.2 days in tonsil. The high tissue concentrations suggest that proposed standard dosage regimens of 500 mg qds on day 1 followed by 250 mg qds for four days, or three daily dosages of 500 mg, will produce tissue concentrations above 3 mg/kg in a variety of tissues. Since these tissue concentrations exceed the MICs of relevant pathogens, these dosage regimens should be effective against respiratory tract and soft-tissue infections. A single 1 g dose may be effective in the treatment of many sexually transmitted diseases.

Adolescent↗

Antibody response reveal differences in oral tolerance to wheat and maize grain protein fractions.

The influence of diet on humoral immune responses to gluten- and maize-derived proteins was examined using ELISA and protein blotting techniques. Mice raised on the maize-based (gluten-free) diet responded well to parenteral immunization with each of six gluten-derived protein preparations (whole gliadin, two omega-gliadin fractions, wheat salt-soluble proteins, a peptic-tryptic digest and a subtilisin digest of gluten), as serum antibody levels increased at least 300-fold in each case. In contrast, mice raised on the wheat-based diet responded poorly to immunization with either whole gliadin or omega-gliadin and were virtually non-responsive to enzymic digest of gluten. Diet had little effect on the magnitude of the antibody response to wheat salt-soluble proteins, with both groups showing a 300-fold increase in titre. Similarly, tolerance to alpha-zeins, the alcohol-soluble proteins of maize, did not occur on either diet. However, some oral tolerance was observed to maize glutelin. The specificity of the various antibody responses was then analysed by immunoblotting. Following immunization with gluten proteins or digests, antibodies from the maize-fed mice bound more or less equally to each of the main gliadin bands and to the glutenins while the mice on the wheat-based diet had antibody specific for omega-gliadin proteins. Serum antibodies from the maize-fed mice, immunized with either alpha-zein or maize glutelin, showed even labelling of the major maize endosperm proteins while antibodies from mice on the wheat diet showed strong labelling of the Mr 27,000 and 58,000 bands. These results show that diet influenced the specificity, as well as the magnitude of serum antibody responses to cereal proteins. In addition, oral tolerance appeared to affect the humoral response to some cereal proteins more than others. Both of these findings have important implications for our understanding of coeliac disease.

Animals↗

Rapid identification of Pasteurella multocida organisms responsible for haemorrhagic septicaemia using an enzyme-linked immunosorbent assay.

Haemorrhagic septicaemia (HS) is caused by specific serotypes of Pasteurella multocida and is one of the major economic diseases of cattle and buffalo in South East Asia. Definitive diagnosis of the disease-causing organism with the available methods is labour intensive and not totally reliable, consequently, an ELISA system to identify P multocida organisms which cause HS was developed. One hundred and twenty-four P multocida isolates were tested, 58 were type strains and 66 were field isolates. Analysis of these strains indicated the assay had a specificity of 99 per cent and sensitivity of at least 86 per cent. The sensitivity could be an underestimate, as five isolates assumed to be false negative reactions may not all be HS-causing strains. The HS ELISA provides a rapid, simple, accurate and inexpensive diagnostic assay for identification of HS causing organisms but does not represent a new typing system for P multocida. This assay will also enable countries to assess the impact of HS more accurately.

Animals↗

Age at death: a three hospital urban, suburban, and small town study.

Comparison of the ages of patients dying at three interrelated St. Louis hospitals in 1988 has been used to study the results of similar high-quality acute hospital care when provided to dissimilar populations. The data presented by the authors leads them to conclude that in this era of mounting health costs, we have not, at least as recently as 1988, realized that an ounce of prevention if worth a pound of cure.

Adult↗

Cloning and characterization of the yeast chaperonin HSP60 gene.

The heat-shock protein, HSP60, is abundant in prokaryotes and eukaryotes and is required in the assembly of specific proteins. We have cloned the Saccharomyces cerevisiae HSP60 gene from a lambda gt11 genomic library using monoclonal antibodies, have obtained its sequence, determined its transcription start point, and shown that it exists as a single copy. The predicted HSP60 contains a mitochondrial target sequence and exhibits striking amino acid sequence similarity to its counterparts in bacteria, plants, and humans. These data indicate a high level of evolutionary conservation and are consistent with the suggestion of evolutionarily conserved function [Hemmingsen et al., Nature 333 (1988), 330-334].

Amino Acid Sequence↗

Distribution of 35S-sulfate within the transseptal ligament of the mouse.

The distribution of 35S-sulfate-labeled macromolecules was examined within three regions of the transseptal ligament: the 1) mesial, 2) middle and 3) distal thirds. Swiss mice, 6 weeks of age, were injected with 35S-sulfate and killed after 1, 6, and 12 hours and 1, 2, 3, 4, 5, and 7 days. Silver grains and cell nuclei were counted on autoradiographs which had been counterstained by the Van Gieson method, and mean counts were analyzed statistically. Analysis of variance revealed no significant differences in mean number of cell nuclei between regions throughout the course of the experiment. 35S-sulfate was rapidly incorporated into the transseptal ligament macromolecules. Grain counts were highest 6 hours after injections: counts were highest over the middle and lowest over the mesial thirds of the ligament. The rate of grain removal was significantly higher in the middle third compared to the mesial or distal thirds (P less than 0.001) and was significantly lower in the mesial third compared to the middle or distal thirds (P less than 0.001). The half-life of labeled macromolecules was significantly greater in the mesial and distal thirds than in the middle third (P less than 0.005). The data demonstrate significantly higher rates of turnover of 35S-sulfate-labeled macromolecules in the middle region of the transseptal ligament. Since cellular density was similar throughout the transseptal ligament, higher turnover rates of 35S-sulfate-labeled macromolecules probably indicate higher rates of cellular activity in this region, possibly a result of tissue remodeling coincident to stresses generated by occlusal forces and physiologic drift of the adjacent teeth.

Animals↗

Effects of hypofunction on the distribution of 3H-proline in the transseptal fibers of the periodontium of the rat.

There is little information concerning the effect of altered occlusal forces on the turnover of collagenous proteins of transseptal fibers of the periodontium. In the present study, hypofunction was induced in rats by extraction of the maxillary teeth, allowing the mandibular teeth to supererupt (hypofunctional side, herein). The contralateral side served as an internal control, although it was likely experiencing occlusal hyperfunction (hyperfunctional side, herein). Untreated animals were also studied (external controls, herein). Animals were injected with 3H-proline and silver grains were counted on radioautographic preparations. The study demonstrated significant differences in the synthesis and degradation of collagenous proteins coincident to altered occlusal function; 3H-proline was most heavily incorporated into the transseptal fibers of hyperfunctional and least rapidly into the external control tissues (P less than .001). Significant differences in grain counts were evident during the first 3 weeks after injections. Collagenous proteins were degraded most rapidly in transseptal fibers of the hyperfunctional and least rapidly in hypofunctional tissues (P less than .001). The study also demonstrated regional variability in the turnover of labeled collagenous proteins, that is, proteins were synthesized and degraded most rapidly in the middle third and least rapidly in the mesial third of the ligament (P less than .001). "Whole" counts (mean of counts over middle, mesial, and distal thirds) were not similar to those of any specific region and could provide erroneous information concerning remodeling of collagenous proteins of transseptal fibers. Transseptal fibers, labeled by the 3H-proline pulse, migrated occlusally with the teeth; new transseptal fibers and bone were formed at the crest of the interdental septum.

Animals↗

A reevaluation of the distribution of 3H-proline in the transseptal ligament of the mouse in radioautography.

It is generally accepted that there is uniform collagen metabolism within the periodontal and transseptal ligaments. The present study suggested regional variations in the incorporation and removal of 3H-proline within the transseptal ligament in radioautography, suggesting variable rates of collagenous protein remodeling coincident with physiological tooth movements. Highest numbers of silver grains were over the middle third of the ligament during both incorporation and removal phases (p less than 0.001). Rates of grain removal were greater in the middle than in mesial or distal thirds (p less than 0.001). The half-life of labeled proteins was significantly less in the middle than in mesial or distal thirds (p less than 0.005). Because there were no significant regional differences in cell numbers, regional variability in grain incorporation and removal within the transseptal ligament likely indicates regional differences in cellular synthetic or degradative activity coincident with remodeling of the transseptal ligament during physiological drift and suggests that the center of this ligament may experience more stress and, thus, remodels more rapidly.

Animals↗

Effect of low-protein diet on alveolar bone loss in streptozotocin-induced diabetic rats.

The present study demonstrated that hyperglycemic diabetic rats fed a low-protein (8%) diet maintained an alveolar bone height similar to controls; in contrast, those fed a standard protein diet (24%) had reduced alveolar bone height (P less than 0.05). Euglycemic diabetic and untreated control rats fed low-protein diets did not have significant differences in alveolar bone height compared to those fed standard protein diets. There was no evidence of gingival or periodontal inflammation or osteoclastic bone resorption at the alveolar crest in any animal studied. Thus, (1) hyperglycemic diabetic rats have significant alveolar bone loss in the absence of periodontal inflammation (P less than 0.001) and (2) this bone loss can be alleviated by diet (P less than 0.05). This data, taken together with previous studies on the effects of low-protein diet on the kidney, suggest that relieving the protein load on the diabetic kidney in poorly controlled diabetics is beneficial to the longevity of that organ, as well as the preservation of alveolar bone surrounding the teeth.

Alveolar Process↗

Alveolar bone of BB/W rats: a morphometric and histochemical study.

The present study reported histochemical changes in alveolar bone glycosaminoglycans (GAG) (using Safranin O) and in interdental bone height in three groups of BB/W rats: diabetic, diabetes prone, and diabetes resistant. Safranin O staining intensity suggested that total GAG levels were highest in diabetic bone (p less than 0.05 compared to diabetes resistant, p less than 0.005 compared to diabetes prone) but not significantly different between diabetes prone and resistant groups. Following chondroitinase AC and ABC digestion, staining reactions suggested that the highest levels of dermatan sulfate were in the diabetes resistant group (p less than 0.001 compared to diabetic, p less than 0.001 compared to diabetes prone) and the highest levels of chondroitin sulfates were in the diabetes prone group (p less than 0.001). Coincidently the mean height of diabetes prone interdental septum was significantly less than that of diabetes resistant or diabetic groups (p less than 0.05). The study suggested that 1) diabetes and "prediabetes" produce significant changes in levels of chondroitin 4, 6, and dermatan sulfates within alveolar bone, 2) in "prediabetic" animals, interdental bone loss occurs prior to the onset of clinical symptoms and in the absence of local irritating factors, the bone height appears to return to normal levels, and 3) there may be a correlation between alveolar bone height and relative levels of dermatan sulfate.

Animals↗

Evaluation of bovine antibody responses to haemorrhagic septicaemia vaccine.

ELISA and immunoblotting techniques were used to examine the humoral immune response to Pasteurella multocida, in bovine sera from Indonesia and Malaysia. Elevated levels of antibody to a crude lipopolysaccharide preparation were found in vaccinated animals. In addition to the response to lipopolysaccharide, antibodies from the vaccinated cattle strongly labelled five to six of the 40 protein bands in this organism.

Animals↗

Staphylococcal carriage and infection in myasthenia gravis patients receiving therapeutic apheresis.

Populations exposed to repetitive needle puncture (such as hemodialysis patients, insulin-dependent diabetics, and parenteral drug abusers) are at increased risk for nasal carriage of Staphylococcus aureus. These groups appear to have increased susceptibility to serious staphylococcal infection as well. We performed a microbiologic survey on a group of patients receiving regular, long-term therapeutic apheresis for myasthenia gravis (MG), and compared the rate of nasal staphylococcal carriage with that found in control groups of MG patients not receiving apheresis and ambulatory general medical patients without known risk factors for staphylococcal carriage. Medical records were reviewed for episodes of significant staphylococcal infection occurring since commencement of apheresis. Nasal S. aureus carriage was found in 9/29 (31%) apheresis patients, 8/30 (27%) MG controls, and 8/30 (27%) general medical controls. No significant difference in frequency of apheresis was noted between carriers and noncarriers. A single episode of S. aureus bacteremia occurred in 95 patient-years of apheresis therapy. We conclude that therapeutic apheresis for MG does not increase the risk of staphylococcal carriage, and that serious infection is infrequent.

Adolescent↗