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Biomedical subjects

R C Johnson

Publications and source records attributed to R C Johnson.

At least 289 records · Page 16Linked to original sources
PubMed1987

Effect of heat or chemical treatment on leptospiral antigens.

The use of bacterins is one of the primary methods in the control of leptospirosis in domestic animals, especially cattle, swine, and dogs. Bacterins have been made using chemically treated or heat-inactivated organisms. Many of the initial studies indicated that the bacterins gave complete protection, whereas later reports found that some inactivation methods resulted in bacterins that protect against death but not against kidney infection and renal shedding. This raised the possibility that some of the antigens had been altered. To investigate this possibility, we heat or chemically treated Leptospira interrogans serovar icterohaemorrhagiae CF-1, L. interrogans serovar canicola Moulton, L. interrogans serovar grippotyphosa SC4397, L. interrogans serovar hardjo Hardjoprajitno, and L. interrogans serovar pomona MLS. Temperatures used to determine alterations in antigens were 50, 56, 80, 100, and 121 degrees C. The chemicals used were formaldehyde, phenol, and thimerosal. Analysis was done using Laemmli gels and Western blots. We found that heating at 50 or 56 degrees C had the smallest effect on antigens, whereas heating at 80 or 100 degrees C caused the appearance of high-molecular-weight bands on Western blots. Heating at 121 degrees C caused almost all the bands to disappear, in both the stained gel and the Western blot. Both phenol and Formalin treatment altered proteins, whereas thimerosal treatment appeared to have little effect compared with heating at 50 degrees C.

Antigens, Bacterial↗
PubMed1987

New infectious spirochete isolated from short-tailed shrews and white-footed mice.

A spirochete with two periplasmic flagella was isolated from the blood or tissues of spleens and kidneys from short-tailed shrews (Blarina brevicauda) and white-footed mice (Peromyscus leucopus) in Connecticut and Minnesota. After inoculation, the shrew-mouse spirochete infected Swiss mice and Syrian hamsters. This spirochete is morphologically and serologically distinct from the species of Treponema, Borrelia, Leptospira, and Spirochaeta examined.

Animals↗
PubMed1987

Seasonal prevalence of Borrelia burgdorferi in natural populations of white-footed mice, Peromyscus leucopus.

Borrelia burgdorferi, the etiologic agent of Lyme disease, was isolated from 111 of 237 Peromyscus leucopus captured during all seasons of the year. Borreliae were cultured from tissues of the spleen (101 mice), left kidney (76 mice), and right kidney (73 mice), from blood (12 mice), and from one fetus. Mice were infected during the winter, when immature Ixodes dammini were inactive. The prevalence of infection during the winter (less than or equal to 33%) was more than twofold lower than that during the summer (ca. 75%), a time when nymphal ticks are abundant. Overwintering, infected mice are reservoir hosts for subadult ticks that begin feeding in early spring. Twenty white-footed mice from which B. burgdorferi was isolated from tissues of spleen or kidney but not from blood were parasitized by larval I. dammini or Dermacentor variabilis which harbored borreliae. We conclude that these mice were infectious to feeding ticks, even though borreliae were not isolated from blood.

Animals↗
PubMed1986

Host protein requirements for in vitro site-specific DNA inversion.

Flagellar phase variation is mediated by a recombination event that occurs at specific sites leading to inversion of a chromosomal segment of DNA. The presence of a 60 bp recombinational enhancer sequence on the DNA substrate molecule results in a 150-fold stimulation in the initial rate of inversion. The protein components required for inversion have been purified. They include the 21,000 dalton recombinase (Hin), a 12,000 dalton host protein (Factor II), and one of the major histone-like proteins of E. coli HU. The dependence of the initial rate of recombination on HU varies with respect to the location of the recombinational enhancer. The role of HU, Factor II, and the enhancer in facilitating site-specific recombination is discussed.

Bacterial Proteins↗
PubMed1986

Growth and lipid composition of rat brain glial cells cultured in lipoprotein deficient serum.

The purpose of this study was to examine the effects of substituting lipoprotein deficient serum (LPDS) for complete fetal calf serum (FCS) in culture media on the growth and lipid composition of cells dissociated from 1 to 2-day-old rat brain. The results show that in FCS cultures DNA, protein and all lipids increase with an increase in the number of days in culture. Substitution of LPDS for FCS in the culture media caused a slower increase in each of these constituents. Esterified cholesterol remained unaltered with time in LPDS cultures but increased continuously in FCS cultures. Substitution of LPDS for FCS reduced the DNA:protein ratio, and unesterified cholesterol:phospholipid ratio but the protein:phospholipid ratio and the proportion of individual phospholipids were not affected. The data indicate that removal of low density lipoprotein (LDL) from serum used in culture media reduces cell proliferation and causes alterations in cellular lipid composition specifically ratio of cholesterol:phospholipids.

Animals↗
PubMed1986

Cholesterol ester hydrolase(s) in mammalian brain: is there a myelin-specific cholesterol ester hydrolase?

The present study compared the properties of cholesterol ester hydrolase(s) in myelin and microsomes from rat, mouse and human brain. The results indicated that the enzyme activity in both myelin and microsomes from rat, mouse and human brain was optimal at pH 6.5 and required Triton X-100 for optimal activity. The enzyme activity in myelin was 3- to 4-fold higher in the presence of Triton X-100 than taurocholate. Addition of phosphatidyl serine enhanced (2 to 4 fold) the hydrolase activity in both myelin and microsomes. The properties of the enzyme in solubilized preparation of myelin were also similar to the properties of the enzyme in partially delipidated and solubilized preparations of microsomes. The activity was again optimal at pH 6.5, required Triton X-100 for optimal activity and was stimulated by phosphatidyl serine. These results indicate that the properties of cholesterol ester hydrolase in myelin are similar to those of the microsomal enzyme and that this is true for the fractions from both human and rodent brain. The data thus lead us to believe that the hydrolase activity in mammalian brain myelin and microsomes may reflect the distribution of a single enzyme in the two fractions rather than two distinct enzymes, one being specific to each fraction.

Animals↗
PubMed1986

Dietary supplementation of undernourished rats with soy or safflower oil: effects on myelin polyunsaturated fatty acids.

Undernourished suckling rats were administered, by gastric intubation, either soy oil (which is rich in both linoleic and linolenic acids) or safflower oil (which is rich in linoleic acid but deficient in linolenic acid) to determine (1) if dietary supplementation would offset the hypomyelination characteristic of the undernourished, developing brain and (2) to compare myelin fatty acids in normal, undernourished, and oil-supplemented rats. Myelin recovery was not increased by supplementation with either oil. The proportions of C22:4 and C22:6 fatty acids were reduced in myelin of the undernourished rats. Undernourished rats supplemented with either soy or safflower oil had higher than normal proportions of polyunsaturated fatty acids (C20:4 and C22:6). The triene-tetraene ratio in the oil-supplemented rats was lower than in normal controls, indicating that the oil-supplemented rats were not deficient in essential fatty acids. No significant differences were observed between the oil-supplemented groups.

Animals↗
PubMed1986

Choline in plasma measured by liquid-chromatography with electrochemical detection.

We have developed a simple, specific, and sensitive method for plasma choline measurement based on the HPLC procedure of Potter et al. (J Neurochem 1983; 41: 188) for the measurement of acetylcholine and choline in neuronal tissue. The effluent from a reverse-phase column is mixed with choline oxidase in a post-column reaction coil to produce hydrogen peroxide which is monitored electrochemically. Plasma samples are prepared by deproteinization with perchloric acid. Choline is recovered quantitatively from the plasma, but an internal standard (homocholine) is added to compensate for any variation in electrode response. Choline can be measured in plasma samples containing less than 1 mumole per litre of plasma; the method response is linear in the 1-20 mumol/L range. Catecholamines and ascorbic acid do not interfere. The chromatography, enzymatic reactions, and electrochemistry all contribute to the specificity of the method.

Alcohol Oxidoreductases↗
PubMed1986

Culturing Borrelia burgdorferi from spleen and kidney tissues of wild-caught white-footed mice, Peromyscus leucopus.

Borrelia burgdorferi was isolated most frequently from tissue of spleen (n = 13) and kidney (n = 10) and less often from blood (n = 5) of wild-caught Peromyscus leucopus. Prevalence of infection tended to be highest at sites where Lyme disease was most common (e.g., 5 of 6 mice were positive in East Haddam, Connecticut). Spirochetes were not isolated in Danbury or New Hartford, areas where this malady is rare. However, in Fairfield, where the disease is also uncommon, 4 of 9 mice were infected. Larval and nymphal I. dammini, containing borreliae, parasitized P. leucopus at all sites where B. burgdorferi was cultured from mice. Borreliae were also detected in D. variabilis feeding on hosts at two of the sites. P. leucopus appears to be an excellent animal to identify focal areas of B. burgdorferi.

Animals↗
PubMed1986

Genetic analysis of Borrelia.

DNA homology studies were performed on Borrelia hermsii, B. duttoni, B. crocidurae and the O. coriaceus spirochete. These organisms had a genus level relationship with each other. B. hermsii was 17 to 53% homologous with B. duttoni, B. crocidurae, and the O. coriaceus spirochetes; B. crocidurae was 17-32% related to B. hermsii, B. anserina and the O. coriaceus spirochete, and the O. coriaceus spirochete was 28-50% related to B. hermsii, B. crocidurae and B. anserina. The O. coriaceus spirochete was 10% homologous with B. burgdorferi, the etiologic agent of Lyme disease. Plasmid DNA was detected in these strains of Borrelia. Southern blot analysis of plasmid DNA from these strains suggested little or no homology with plasmid DNA from B. hermsii used as a probe. A plasmid of 9.2 kilobases was seen in B. burgdorferi strain CT1. This plasmid was shown to be a tandem repeat with unit length of 4.6 kilobases. Sequences corresponding to this plasmid were detected in several fresh tick isolates from North America, and were not seen in any animal, human or European isolates surveyed. The role in the biology of the spirochete played by this plasmid is unknown.

Animals↗
PubMed1986

Involvement of birds in the epidemiology of the Lyme disease agent Borrelia burgdorferi.

Borrelia burgdorferi, the causative agent of Lyme disease, was isolated from the liver of a passerine bird, Catharus fuscescens (veery), and from larval Ixodes dammini (tick) feeding on Pheucticus ludovicianus (rose-breasted grosbeak) and Geothlypis trichas (common yellowthroat). In indirect immunofluorescence antibody tests, isolates reacted with polyclonal and monoclonal (H5332) antibodies. Studies on the DNA composition of the veery liver isolate and the strain cultured from an I. dammini larva indicated that both were B. burgdorferi and not Borrelia anserina or Borrelia hermsii. The veery liver isolate infected hamsters and a chick. In contrast, B. anserina infected chicks but not hamsters. B. burgdorferi is unique among Borrelia spp. in being infectious to both mammals and birds. We suggest that the cosmopolitan distribution of B. burgdorferi may be caused by long-distance dispersal of infected birds that serve as hosts for ticks.

Animals↗
PubMed1986

Passive immunization of hamsters against experimental infection with the Lyme disease spirochete.

Hamsters passively immunized with as little as 0.0125 ml of immune rabbit serum (indirect fluorescent-antibody titer, 1:8,192) were protected from challenge with 1,000 50% infective doses of Borrelia burgdorferi. Cross-protection studies with Minnesota and Connecticut isolates of B. burgdorferi indicated no major differences in their capacity to elicit mutually protective antibodies in rabbits.

Animals↗
PubMed1986

Active immunization of hamsters against experimental infection with Borrelia burgdorferi.

The immunogenicity of a single dose of a whole-cell vaccine of inactivated Borrelia burgdorferi HSF (human spinal fluid isolate) was evaluated. Syrian hamsters were vaccinated subcutaneously and challenged by the intraperitoneal injection of 1,000 50% infectious doses of B. burgdorferi HSF 30 or 90 days postvaccination. Animals were sacrificed 14 days after challenge, and the kidneys and spleens were examined for spirochetes by cultural procedures. At 30 days postvaccination, 86 to 100% protection against infection was achieved in hamsters receiving 50 and 100 micrograms (dry weight) of vaccine. Protection was decreased to 60% with 25 micrograms of vaccine and was absent with 10 micrograms of vaccine. Resistance to infection decreased to 25, 40, and 5% for the 100-, 50-, and 25-micrograms vaccine doses, respectively, at 90 days postvaccination.

Animals↗
PubMed1986

Peromyscus leucopus and Microtus pennsylvanicus simultaneously infected with Borrelia burgdorferi and Babesia microti.

Borrelia burgdorferi, the etiologic agent of Lyme disease, and Babesia microti, the causative agent of human babesiosis, were isolated from 71 and 57%, respectively, of 14 specimens of Peromyscus leucopus and Microtus pennsylvanicus collected from Prudence and Patience Islands, R.I. Both pathogens were isolated from five individual rodents. The presence of these two infectious organisms in the same mammal suggests that individual larval Ixodes dammini may ingest both pathogens and subsequently transmit them in the nymphal stage.

Animals↗