Ultrastructure of the human intra-articular disc of the temporomandibular joint.

The ultrastructural appearance of the human intra-articular disc (IAD) was investigated in three discs that had been surgically removed due to disease from three female patients aged 47, 50, and 54 years of age. Regions of the IAD were selected from central areas that appeared to be least affected by disease. Sections were fixed in 2.5 per cent glutaraldehyde in 0.1 M phosphate buffer, pH 7.3 immediately after surgery. The regions examined showed no obvious signs of inflammation. The cells showed moderate amounts of the intracellular organelles associated with protein synthesis and secretion, and possessed considerable amounts of microfilamentous material, thus resembling those described in other mammals. Despite the large number of cells examined, only one cell showed evidence of a chondrocyte-like morphology in that it possessed an incomplete pericellular zone of microfilamentous material separating the cell membrane from the adjacent collagen bundles of the extracellular matrix (ECM). Thus, on morphological grounds, fibrocartilage was virtually non-existent in the specimens examined. The mean collagen fibril diameter was 43.9 nm and the fibril diameter distribution was not unimodal. Although the majority of fibrils had a relatively small diameter, two of the three specimens possessed many fibrils with diameters of over 100 nm, this being consistent with tissue subjected to tension. The mean area of a fibre bundle occupied by collagen (as opposed to the ground substance) was approximately 56 per cent.

Age changes in the cells of the intra-articular disc of the temporomandibular joints of rats and marmosets.

Cells in the intra-articular disc of the temporomandibular joint were studied ultrastructurally at three different ages to investigate any age changes. Rats aged 2, 15.5 months, and 2.5 years, and marmosets aged 21 months, 7 years, and between 10.5 and 14 years were studied. In the first two age groups of the rat and the first of the marmoset, the cells were generally rounded and had moderate amounts of rough endoplasmic reticulum and other organelles associated with protein synthesis and secretion. Many cells had conspicuous amounts of microfilamentous material and cell membranes were closely applied to the collagen fibrils of the extracellular matrix. Occasionally, a narrow, irregular space containing microfilamentous material lay adjacent to the cell membrane. In the 2.5-year-old rats and the two older age groups of marmosets, cells with chondrocyte-like morphology were present. These cells were surrounded by a conspicuous pericellular matrix devoid of collagen fibrils and composed of microfilamentous material embedded in an amorphous ground substance. They resembled cells described in fibrocartilage from other sites, but differed from chondrocytes in hyaline cartilage by lacking a pericellular capsule. Thus, rats and marmosets both show cellular age changes in the intra-articular disc of the mandibular joint, which can be considered as changing from fibrous to fibrocartilaginous with age, a condition similar to that reported in humans.

The ultrastructure of the intra-articular disc of the temporomandibular joint, with special reference to fibrocartilage.

Cells in the intra-articular disc of the temporomandibular joint of the rat, guinea pig, rabbit, ferret, marmoset and sheep were studied at the ultrastructural level. The cells were generally rounded in outline and possessed moderate amounts of roughened endoplasmic reticulum and other organelles associated with protein synthesis and secretion. No intracellular collagen profiles were observed. Many of the cells possessed conspicuous amounts of microfilamentous material. Cell membranes in the rat, guinea pig, rabbit, ferret and sheep were closely applied to the collagen fibrils of the extracellular matrix. Occasionally in these animals, a narrow, irregular space containing microfilamentous material surrounded the cell membrane. Many cells in the marmoset differed from this description in being completely surrounded by an obvious pericellular matrix devoid of collagen fibrils and being comprised of microfilamentous material embedded in an amorphous ground substance. These chondrocyte-like cells in the intra-articular disc of the marmoset differed from chondrocytes in hyaline cartilage by lacking a pericellular capsule.

Enantiomers of flurbiprofen can distinguish key pathophysiological steps of NSAID enteropathy in the rat.

BACKGROUND: Non-steroidal anti-inflammatory drugs (NSAIDs) cause gastrointestinal damage by a non-prostaglandin (PG) dependent "topical" action and by inhibiting cyclooxygenase. AIMS: To discriminate between these two effects by studying some key pathophysiological steps in NSAID enteropathy following administration of (R)- and (S)-flurbiprofen, the racemic mixture, and an uncoupler, dinitrophenol. METHODS: The effects of dinitrophenol, racemic, (R)-, and (S)-flurbiprofen on mitochondria were assessed in vitro and on key pathophysiological features of small intestinal damage in vivo (ultrastructure by electron microscopy, mucosal prostanoid concentrations, intestinal permeability, inflammation, and ulcer count) in rats. RESULTS: All the drugs uncoupled mitochondrial oxidative phosphorylation in vitro, caused mitochondrial damage in vivo, and increased intestinal permeability. Dinitrophenol and (R)-flurbiprofen caused no significant decreases in mucosal prostanoid concentrations (apart from a decrease in thromboxane (TX) B2 concentrations following (R)-flurbiprofen) while racemic and (S)- flurbiprofen reduced mucosal prostanoids significantly (PGE, TXB2, and 6-keto-PGF1alpha concentrations by 73-95%). Intestinal inflammation was significantly greater following administration of (S)-flurbiprofen and racemate than with dinitrophenol and (R)-flurbiprofen. No small intestinal ulcers were found following dinitrophenol or (R)-flurbiprofen while both racemic and (S)-flurbiprofen caused numerous ulcers. CONCLUSIONS: Dinitrophenol and (R)-flurbiprofen show similarities in their actions to uncouple mitochondrial oxidative phosphorylation in vitro, alter mitochondrial morphology in vivo, increase intestinal permeability, and cause mild inflammation without ulcers. Concurrent severe decreases in mucosal prostanoids seem to be the driving force for the development of severe inflammation and ulcers.

Ultra-structure and localisation of formazan formed by human neutrophils and amoebae phagocytosing virulent and avirulent Legionella pneumophila.

Legionella pneumophila (LP) strains of differing virulence were incubated with a solution of nitroblue-tetrazolium (NBT) at a concentration of 1 mg.ml-1 in the presence of Acanthamoeba polyphaga or human polymorphonuclear neutrophils (PMN). Reduction of NBT to formazan occurred at a faster rate in the presence of virulent strains. Reduction appeared to be temperature dependent; at 37 degrees C the reaction rate was higher than at 20 degrees C. On microscopic examination, deposits of formazan around Legionella cells were observed inside amoebae similar to those deposited in human neutrophils. Electron microscopy revealed electron-dense particles surrounding virulent legionellae, which appeared to be associated with formazan formation. Formazan formation inside amoebae may suggest the presence of a respiratory burst against LP, which is more intense with virulent strains.

Immunogold labeling for electron microscopy.

Colloidal gold immunocytochemistry was first introduced by Faulk and Taylor (1), and has rapidly become a major technique in electron microscopy, covering many aspects of biological research. The rapid expansion of this technique in electron microscopy is first the result of its simplicity compared with other labeling techniques, and secondly, the result of the properties of the gold probes themselves. These probes are usually 3-15 nm in diameter and coated with immunologically active proteins. They have the advantages of being very electron dense, giving a characteristic appearance that cannot be confused with other biological structures; they are highly sensitive, producing very specific labeling of both monoclonal and polyclonal antibodies; they are also permanent, nonhazardous, and can be easily quantified.

Phospholipids in plant and animal chromatin.

Isolated hepatic nuclei and hepatic chromatin have been analysed for their DNA, RNA, protein and phospholipid content. The protein/DNA ratio is 3 for nuclei and 1.95 for chromatin extracted from Triton X-100 treated nuclei. The phospholipids, (2.36 +/- 0.91 (S.D.) per cent of the total nuclear material), are lost during the chromatin preparation mainly during the Triton X-100 washings of the nuclei. Nevertheless, 10 per cent of the total nuclear phospholipids remain bound to the chromatin. The comparative analysis of both nuclei and chromatin shows a difference in phospholipids and fatty acid composition. Thus, the chromatin-associated phospholipid cannot be attributed simply to contaminating nuclear membrane. This is supported by the autoradiographic study of semi-thin sections of interphase nuclei from root apices of Vicia faba in which [3H] ethanolamine is clearly localized in the chromatin and nucleolar regions of the nuclei.

The effect of chymotrypsin on the development of Dictyostelium discoideum.

Development of the cellular slime mould Dictyostelium discoideum strain NC4, in the presence of alpha-chymotrypsin (3 mg/ml) is reversibly arrested at the tight aggregate stage (10/12 h). Pronase has a similar effect, but trypsin only retards normal development by about five hours. Normally developing cells are susceptible to alpha-chymotrypsin if they are transferred into its presence at any time up to the tight aggregate stage (10-12 h). Transfer after this stage does not affect the appearance of fruiting body structures in the normal time (24 h). Electron microscopy showed the ultrastructure of alpha-chymotrypsin-blocked aggregates after starvation for 24 h to be consistent with a block at 10-12 h of normal development. Poorly developed prespore vacuoles, having thin incomplete walls and a paucity of electron-dense material, are present in some cells. No angular vacuolated cells characteristic of stalk cells are visible. Fruiting bodies formed in the presence of a alpha-chymotrypsin, either as minority structures when the enzyme is added before 10-12 h of normal development, or as the majority structures on later enzyme addition, were found to be abnormal. Normal stalks were formed but the spores were immature. Prespore vacuoles were present, though disrupted, and the cells were not encapsulated by spore walls. The electronegativity of intact slime mould amoebae was significantly reduced, and material containing L-[6-3H]-fucose and [1-14C]leucine was removed from the cell surface on alpha-chymotrypsin treatment. Few plasma membrane proteins were affected, however, and staining of polyacrylamide gels for glycopeptides using Con A-peroxide binding also showed little change.