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Biomedical subjects

K Fujisaki

Publications and source records attributed to K Fujisaki.

At least 109 records · Page 6Linked to original sources

Molecular cloning and immunological analysis of immunodominant piroplasm surface proteins of Theileria sergenti and T. buffeli.

cDNA libraries of Theileria sergenti and T. buffeli piroplasms were constructed in lambda gt11 and screened with rabbit anti-piroplasm sera. A major antigen of T. sergenti (33 kDa) and that of T. buffeli (34 kDa) was identified from the recombinant phages by using recombinant antigen-selected monospecific antibodies. The reactivities of the cloned proteins with rabbit antisera, infected calf sera and mouse monoclonal antibody suggested that the 33 and 34 kDa proteins expressed species-common and species-specific epitopes. The DNA probes from these recombinant clones showed species-specific hybridizations in Southern blotting with genomic DNA from piroplasms. These results indicate that the Japanese T. sergenti can be distinguishable from the Australian T. buffeli with regard to a polymorphism of the major immunodominant proteins of piroplasm.

Animals↗

Purification of merozoites of Theileria sergenti from infected bovine erythrocytes.

A simple and rapid method for purifying merozoites of Theileria sergenti from infected bovine erythrocytes was developed. Infected erythrocytes were lysed by the use of a cytolytic toxin produced by Aeromonas hydrophila and the lysate was subjected to ultracentrifugation in a Percoll discontinuous density gradient. Pure and morphologically intact merozoites free of erythrocyte membrane were recovered from a band formed at the interface of 40% and 60% (by vol.) Percoll solutions. In this merozoite fraction, contamination of erythrocyte membrane proteins was not detected as examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Aeromonas↗

Protein analysis of Theileria sergenti/buffeli/orientalis piroplasms by two-dimensional polyacrylamide gel electrophoresis.

Proteins of the piroplasms of Theileria sergenti, T. buffeli and T. orientalis were analysed by two-dimensional polyacrylamide gel electrophoresis. Protein spot patterns of T. buffeli and T. orientalis were identical except for a few minor proteins, whereas spot patterns of two T. sergenti stocks were differentiated from those of T. buffeli and T. orientalis by a characteristic set of proteins including a major protein of molecular weight 33-34 kDa. This result indicates that Japanese T. sergenti can be phenotypically distinguishable from European and Australia Theileria species; T. orientalis and T. buffeli.

Animals↗

Colonic mucosal interleukin-6 in inflammatory bowel disease.

Interleukin-6, a cytokine produced by various cell types, has a major role in inflammatory and immunological reactions. To define its potential role in inflammatory bowel disease, its concentrations in endoscopic biopsy samples from patients with ulcerative colitis and Crohn's disease were measured. The involved colonic mucosa from active disease was found to contain significantly larger amounts of interleukin-6 than that from inactive disease or normal controls. Colonic mucosal interleukin-6 levels correlated well with the grade of macroscopic inflammation, especially in patients with ulcerative colitis. The levels of interleukin-6 decreased in parallel with clinical improvement following the start of therapy in patients with both forms of inflammatory bowel disease. Mucosal interleukin-6 is thus concluded to accurately reflect the degree of colonic inflammation and may be importantly associated with inflammatory and immunological phenomena seen in inflammatory bowel disease.

Adult↗

Infection rates of Theileria sergenti in Haemaphysalis longicornis ticks collected from the field in Japan.

For a period of one year Haemaphysalis longicornis ticks were collected in a pasture from vegetation by dragging flannel cloth and from soil samples at monthly intervals. Nymphal ticks were assessed for Theileria infection. Salivary glands were stained with methylgreen-pyronin and examined for parasite masses. Nymphal H. longicornis infected with Theileria sergenti were found in all samples during 12 months including the winter. After shortening the prefed period on rabbits to 24 hours, the parasite masses could be detected in the salivary glands of nymphs collected in the grazing season, from May to October, while no parasite masses were detected in other season. It was suggested that the environmental factors in the grazing season might influence on the maturation of parasites in the salivary glands of ticks.

Animals↗

Detection of the schizont stage in experimentally infected cattle with Japanese Theileria sergenti.

Calves infected with a sporozoite suspension of the Ikeda strain of Theileria sergenti were examined in attempts to detect the schizont stage. A suspension equivalent to the contents of 70-200 ticks was inoculated subcutaneously into 10 splenectomized calves. Schizonts and micromerozoites were found in the prescapular lymph nodes of 5 calves about 8 days after the sporozoite inoculation. Schizonts detected at the time of nuclear division had large and diffuse nuclei, and many of the schizonts were very large and extracellular in lymph node smears. The schizonts were also detected by the direct fluorescent antibody staining.

Animals↗

Correlation between the infection rate of the vector tick, Haemaphysalis longicornis and the parasitaemia of cattle infected with Theileria sergenti.

The infection rate of Theileria sergenti in salivary glands of nymphal Haemaphysalis longicornis which had dropped from cattle showing different piroplasm parasitaemias was examined by the methyl green-pyronin staining method. The results suggested that there was some correlation between parasitaemia and the subsequent infection rate in salivary glands of ticks. It seemed possible that the parasitaemia in cattle, especially the number of parasitized erythrocytes in blood imbibed by ticks, might be an important influence on the number of infected acini of ticks.

Animals↗

Development of precipitating antibody in chickens experimentally infested with northern fowl mite, Ornithonyssus sylviarum (Acari: Macronyssidae).

In order to examine the immune response of chickens to different population levels of mites, a microscopic slide modification of the Ouchterlony double-gel diffusion technique was adopted for examination of circulating antibody against the extract of northern fowl mite, Ornithonyssus sylviarum. Precipitating antibodies were detected in all the chickens infested with the mite. One to three clearly defined precipitation lines appeared in almost all the serum samples of infested birds. Titers of antibody correlated with population levels of the mite on chickens, and no differences in antibody development of hens and roosters were distinguished. These results suggest that the titration of precipitating antibodies appears to be useful for the assessment of mite population levels on chickens.

Animals↗

Comparison of effective toxicant biotransformation by autochthonous microorganisms and commercially available cultures in the in situ reclamation of abandoned industrial sites.

Biotransformation and biodegradation of polycyclic aromatic hydrocarbons by the subsurface autochthonous microflora collected from an abandoned petroleum refinery site was investigated. In addition, the inoculation of a commercially available blend of bacterial cultures, with known ability to degrade polycyclic aromatics, was evaluated. This supplemental addition of select microorganisms has been referred to by commercial interests as "bioaugmentation". Their biodegradative potential was evaluated using laboratory mesocosms (simulations) containing a predetermined optimal waste loading rate based on % oil and grease, mixed with predetermined optimal loading rates of clay and river silt materials. The waste consisted primarily of aliphatic and aromatic hydrocarbons found in buried sludges and abandoned lagoons. All mesocosms received nutrient amendment without additional energy sources. Experimental mesocosms were inoculated with adapted indigenous microflora and/or commercial strains. Microbial ATP, microbial diversity, and related enzyme assays were used to establish the detoxification efficiency of the experimental microflora. Quantitative toxicant concentrations and transformations were documented by GC/MS data. Information will be presented mainly on the kinetics of toxicant biotransformation processes to identify "bioaugmentation" contribution and relevancy in the recovery of abandoned polluted sites.

Bacteria↗