The adherence of Porphyromonas gingivalis (P. gingivalis) to the periodontal tissue may be an initial step in the pathogenesis of periodontal disease, and fimbriae is believed to play an important role in such processes. However, the heterogeneity of the size, sequence and antigenic reactivity of the fimbriae have been reported. The aim of the present study was to investigate the heterogeneity of P. gingivalis strains on the fimbrillin gene locus by RFLPs analysis.
Fourty-seven P. gingivalis strains including 5 reference strains were used in this study. The plasmid (pUC13Bg 12.l) with the insert of fimA381 was modified with bisulfite and used as the probe. The genomic DNAs from P. gingivalis were digested with the restriction endonuclease Sac I and Pst I, electrophoresed, transferred and hybridized with the DNA probe.
All Sac I digests generated one major band and the band size was almost the same (ca 2.5-kb), except strain W50. The Pst I digests showed one or two major bands and could be divided into 9 groups based on the band patterns. Moreover, isolates from one patient showed different band patterns
By RFLPs analysis, genetic heterogeneity seems to exist within the fimbrillin gene locus of P. gingivalis strains. Such genetic heterogeneity may reflect the previously repo1ted difference of P. gingivalis fimbriae and moreover a single patient could be infected with more than one genotype of P. gingivalis.
Nitric oxide (NO) is a free radical that has been recently recognized as a neuronal messenger molecule. In order to understand the way in which NO functions in the central nervous system (CNS), it is important to identify the NO-generator and NO-target cells in the brain. I measured firstly the distribution of NO synthase in the brain, which catalyzes L arginine to form NO, by the measurement of citrulline formation that is also synthesized from L-arginine together with NO in equal molar bass. In the brain of adult rat, the most potent activity of NOS was apparent in the cerebellum, next in the olfacto1y bulb and medium in the cerebrum. Further, in the presence of NADPH and Ca2+, NOS activity was detected in the neuron cultures derived from the cerebrum of fetal rat. Astrocytes, one type of glia, prepared also from the cerebrum of fetal rat, appeared to have a small but significant NOS activity. As astrocytes possess a high amount of cytosolic guanylate cyclase that is known to be activated by NO, the changes in the intracellular cGMP levels in the astrocytes were measured as another index of NO formation. The treatment of astrocytes with NOS inhibitor caused the suppression of the intracellular cGMP levels. These results indicate that NO is definitely produced by astrocytes. In addition, in the blood vessel system of the brain, although NOS has been thought to be localized in the endothelial cells of only larger vessels, NOS activity was also observed in the microvessel endothelial cells of the cerebrum of both adult and fetal pig. These data suggest that neuronal cells may be the major site of NO generation in the brain, and that the NOS is a constitutive type. The data also suggest that astrocytes can also express constitutive NOS, although the potency is not so large. Microvessel endothelial cells of the brain are also one of the sources of NO. The NO produced by these cells increases the cGMP levels in the astrocytes and may affect some physiological and/or pathophysiological events in the brain.
In a series of studies to investigate the structural features of the biological crystals, such as the tooth and bone, using an electron microscope, we examined the ultrastructure of the enamel, dentin, and bone crystals at near atomic resolution and showed the configuration of the hydroxyapatite structure through the cross and longitudinal sections of the crystals.
Thereafter, based on the results of the observations by the authors of the ultrastructure of the tooth and bone crystals, thinking that it might be possible to conduct direct three-dimensional observation of the configuration composing the unit cell of the hydroxyapatite crystals, we conducted a study on this. These results indicated that it was possible to sterically observe the configuration of the hydroxyapatite structure composing the enamel crystal.
The materials used for this study were the middle layer of the noncarious enamel from the freshly extracted human erupted permanent molars. The small cubes of the enamel were fixed in glutaraldehyde and osmium tetroxide and embedded in epoxy resin using the routine methods. The ultrathin sections were cut with a diamond knife without decalcification and were examined with the HITACHI H-9000 H type transmission electron microscope operated at 300 kV. Each crystal was observed at an initial magnification of 500,000 times and at the final magnification of 10,000,000 times and over.
We sincerely believe that the electron micrographs shown in this report are the first to show three-dimensionally the configuration of the hydroxyapatite structure composing the crystal in the cross and logitudinal sections of an enamel crystal.
In a series of studies to investigate the basic structural features and characteristics of the biological apatite crystals, using an electron microscope, we examined the ultrastructure of the human enamel, dentin, and bone crystals at near atomic resolution and showed the configuration of the hydroxyapatite structure through the cross and longitudinal sections of the crystals.
Subsequently, based on the results of the observations by the authors of the ultrastructu1 e of the tooth and bone, using the same approach, we have been able to directly examine the images of the lattice imperfections in the human enamel, dentin, and bone crystals, such as the point defect structures and dislocations in the crystals.
In this report, we describe the image of the point defect structures and line defect structures obtained, using the same approach from the sections of the human enamel, dentin, and bone crystals. The materials used for this study were the noncarious enamel and dentin from the freshly extracted human erupted lower first molars, and bone tissue obtained from the alveolar compact bone. The small cubes of the material were fixed in glutaraldehyde and osmium tetroxide and embedded in epoxy resin using the routine method s. The ultrathin sections were cut with a diamond knife without decalcification. The sections were examined with the HITACHI H-800 H and H-9000 types of transmission electron microscopes operated at 200 kV and 300 kV. Each crystal was observed at the initial magnification of 300,000-500,000 times and at the final magnification of 10,000,000 times and over.
We sincerely believe that the electron micrographs shown in this report are the first to show the images of the lattice imperfections in the human enamel, dentin, and bone crystals, such as the point defect and line defect structures, at near atomic resolution.