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The occlusal fissure of unerupted third molars as an experimental caries model in man.

The aim of the present investigation was to establish an experimental model for the study of fissure caries development under conditions as similar as possible to the in vivo situation. Twenty-one blocks (2 X 2 X 3 mm) constituting part of the occlusal fissure were made from unerupted human third molars, and implanted for 60 or 90 d in mandibular molars of students. One milliliter of a 50% sucrose solution was applied to the fissures twice a day. After removal, sections were examined microradiographically in polarized light and in the electron microscope. As a control, the morphology and enamel structure of fissures from 10 unerupted third molars were studied. After 90 d a subsurface, radiolucent caries-like lesion was observed in two specimens only. When examined in air in polarized light, however, all sections exhibited caries-like lesions symmetrically located around the fissures. The 60-d material showed similar lesions, but not as frequently as the 90-d specimens. In the experimental material processed for electron microscopy revealed that the fissure plaque mainly consisted of cocci and small rods. Most often the bacteria were separated from the enamel surface by a pellicle.

Dental Caries

Antibacterial functions of macrophages in experimental protein-calorie malnutrition. I. Description of the model, morphologic observations, and macrophage surface IgG receptors.

An experimental protein-calorie malnutrition was produced in weanling Sprague-Dawley rats. The model resembles human malnutrition with respect to weight loss, inanition, angular stomatitis, anemia, lymphopenia, hypoproteinemia with hypoalbuminemia, and marked thymic involution. In addition, systemic invasion by gram-negative rods was documented. However, no edema was produced, and animals did not survive for longer than six weeks on the protein-deficient diet. One percent glycogen was found to be a satisfactory nonprotein stimulus for induction of a peritoneal exudate consisting primarily of young macrophages. Electron microscopy showed that morphologic events of phagocytosis and degranulation proceeded normally in macrophages from protein-deficient animals. In addition, cell surface receptors for IgG were preserved under these experimental conditions. These data indicate that weanling rats may be employed as a small animal model for servere, fulminant protein-calorie malnutrition in humans.

Animals

[Congenital desproportion of various types of muscle fiber, with relative small size of type I fibers. Morphological documents on muscle biopsies in 3 members of the same family].

In two sisters with a neo-natal hypotonia, muscle biopsies demonstrated as main pathological feature a disproportion in size between the two types of muscle fibers defined according to their myofibrillar ATPase activity. Type I fiber mean diameter was at the lower limit of the normal values, and type II fibers were larger than normal. Their father's biopsy also showed an abnormal smallness of the type I fibers, with a bimodal distribution. By electron microscopy, the small type I fibers did not reveal any significant abnormality in children's biopsies. In father's biopsy, there was an abnormal degree of filamentary interchange between contiguous myofibrils and a few stacks of rods in the type I fibers. These three cases demonstrate the familiar character of the disorder. The relationship of this new entity with the other congenital myopathies is controversial, as a similar congenital fiber type disproportion, has been found in association with different ultrastructural changes. Several data favour an insufficient development of the type I fibers rather than an atrophying process. The mechanism of this "hypotrophy" remains unknown.

Adenosine Triphosphatases

Ultrastructure and cytokinetics of leukemic myeloblasts containing giant granules.

Leukemic myeloblasts containing abnormal granules were studied with ultrastructural, cytochemical, and thymidine labeling techniques to evaluate defects in granulogenesis and proliferation. Giant granules (1 to 3 micron in diameter) and Auer rods were observed in leukemic cells from two patients, and only rarely were both abnormal granule types observed in the same cell. The lysosomal origin of these abnormal granules was demonstrated by their content of peroxidase, esterase, and anionic glycoconjugates. Fusion of small dense granules (less than 0.2 micron in diameter) appeared to be increased in cells containing Auer rods and/or giant granules, but fusion of intact primary granules (0.2 to 0.4 micron in diameter) and sequestration of cytoplasmic contents were observed only in giant granules and not in Auer rods. Although the small granules that fused to form giant granules and Auer rods appeared similar, there was no evidence for transformation of giant granules into Auer rods. In one patient, cells with abnormal granules could easily be distinguished from the larger population of cells that lacked abnormal granules. The perturbation of these two distinct populations by chemotherapy was evaluated with thymidine labeling experiments. A high percentage (2- or 3-fold greater) of the abnormally granulated myeloblasts incorporated tritiated thymidine when compared to myeloblasts without abnormal granules in the same specimen. This difference could have resulted from an underlying metabolic defect which affected both granulogenesis and cell division. These results demonstrate that the formation of giant granules in leukemic cells is morphologically similar to that observed in the Chediak-Higashi syndrome and that leukemic cells with abnormal granules may differ cytokinetically from uninvolved leukemic cells.

Cell Division

Extraocular muscles: light microscopy and ultrastructural features.

Thirty extraocular muscles (EOM) from 20 patients were evaluated by light microscopy (LM), electron microscopy (EM), and enzyme histochemistry (EZH). Twenty-one EOM were obtained from 13 patients with strabismus, 9 EOM from 4 patients undergoing eye surgery for other reasons and from 3 autopsy cases. One mum thick sections revealed marked variation in muscle fibre shape and size and in myofibrillar structure; also noted were small, hypertrophied, whorled, and ringbinden fibres. Dense and granular material in the central portion of some fibres and sarcomere disruption in 2--3 mum sections was observed. EZH revealed the absence of the classical mosaic pattern usually found in skeletal muscles. ATPase studies were inconsistent and did not correlate with the expected reciprocal activity of NAD-H diaphorase, particularly on the large fibres. Ultrastructural features consisted of vacuoles within myofilament bundles, "smearing" of Z bands, and "nemaline rods". Occasional myelin figures and lipid-like droplets were observed in subsarcolemmal spaces, associated with scattered clusters of glycogen granules. Abnormal mitochondria and subsarcolemmal inclusions of dense and granular material were conspicuous. "Leptomeric" profiles, "Zebra bodies", or "striated bodies" were noted in 8 EOM's, and an Hirano body was found in 1. The intramuscular nerves contained structures resembling "Luse bodies" in 7 cases. These observations suggest that EOM from individuals with and without strabismus possess unique structural characteristics suggestive of developmental and morphological disarrangement of contractile elements. Some of these changes might play a role in the pathogenesis of strabismus and in the development of clinical symptoms. These features are significantly different from striated skeletal muscle. Therefore the criteria used in the pathological evaluation and diagnosis of skeletal muscle disorders cannot be unequivocally applied to EOM investigations. These data establish the necessity to determine histological norms, ultrastructural patterns, and develop new enzyme histochemistry criteria for the evaluation of EOM. Only then can an acceptable comparison of EOM and skeletal muscle be made.

Adenosine Triphosphatases

Sensitivity of toad rods: Dependence on wave-length and background illumination.

1. There are five morphological types of photoreceptors in the retina of the toad, Bufo marinus: red and green rods, single cones, and the principal and accessory members of double cones. The largest and most abundant of these is the red rod. 2. Intracellular recordings were used to investigate the dependence of the sensitivity of red rod responses on wave-length and background light. 3. The spectral sensitivity of dark-adapted and moderately light-adapted red rods can be satisfactorily fitted with the absorbance spectrum of the red rod photopigment. There are no significant contributions to red rod responses from cones or green rods. 4. In contrast, L-type horizontal cells, whose responses are dominated by input from the red rods near threshold, can be shown also to receive input from cones. 5. Steady background light produces a response in the red rods consisting of an initial hyperpolarization, followed by a decay of potential to a steady-state plateau level. The slow decay of response amplitude is accompanied by an increase in sensitivity to increment test flashes. 6. The increment sensitivity at steady-state decreases with increasing background intensity according to a modified Weber-Fechner relation. The dependence of increment sensitivity on the wave-length of the background light can be predicted by the red rod spectral sensitivity, showing that cones do not influence the light adaptation of rods. 7. At a background [corrected] intensity of 11-5 log equivalent quanta cm-2sec-1, sensitivity begins to deviate from the Weber-Fechner relation. In background light one log unit brighter, the rods are completely saturated. 8. Small responses having the spectral sensitivity of cones can be recorded from saturated rods. These potentials have a prominent off response whose wave form resembles the d-wave of the e.r.g. 9. A comparison of the increment--sensitivity curves of single receptors shows that rods are light-adapted by backgrounds one thousand times dimmer than those which affect cones. The increment--sensitivity curves of rods and cones cross, so that single cones become more sensitive than single rods even before the rods begin to saturate.

Adaptation, Ocular

Bacterial R-bodies with common morphologies and unrolling dynamics are phylogenetically scattered, indicating extensive lateral gene transfer and wide application potential.

Refractile bodies (R-bodies) of gram-negative bacteria are large proteinaceous assemblies, rolled up in the form of an Archimedean spiral. They exhibit rapid rod-like reversible extension in the micrometer range when cued by chemical environmental triggers and have potential for synthetic biology and biochip applications. Initially described for the Paramecium endosymbionts Caedibacter taeniospiralis and Caedimonas varicaedens, R-bodies have since been discovered in many classes of Pseudomonadota, both in endosymbionts and in non-endosymbionts. However, despite the fact that the genetics and morphologies, as well as the unrolling kinetics of R-bodies from different species, show considerable diversity, no recent study has integrated these aspects into a single framework. The latter would be advantageous for the creation of an R-body biotechnology toolbox, where different properties determine the application area. Here, we have examined the R-bodies from six different Pseudomonadota, comprising both phylogenetically diverse endosymbionts and non-endosymbionts. Comparison of the morphologies of the rolled-up and unrolled forms, obtained using electron microscopy and high-quality images, to their corresponding genetic data indicates that extensive lateral gene transfer has occurred, which confounds a common framework based on these data. However, we have also studied the R-body extension and retraction kinetics using high frame-rate light microscopic video recordings, where we show for the first time that R-bodies can be classified into two classes, showing "fast burst" or "slow" acid-induced extension kinetics, respectively. We propose that this criterion may, in fact, be the most useful for the choice of an R-body tool for biotechnological purposes.IMPORTANCER-bodies are unique proteinaceous macromolecular structures capable of massive reversible extension in response to external environmental triggers without the input of chemical energy. They comprise only a few small polypeptides, which makes them potentially highly amenable to tuning via genetic engineering, as well as being exceptionally stable. These properties would be highly desirable in biotechnology and synthetic biology, as well as in biochip applications, where a controlled mechanical extensor might play an integral part in a nanoscale molecular machine. So far, only R-bodies from a single species, Caedibacter taeniospiralis, have been characterized extensively. However, in recent years, genomic information has revealed that a panoply of R-bodies are widely distributed among gram-negative phyla, although studies have generally not included morphological data. This study brings these two areas together to provide a holistic overview of the field and also reveals new insights into key dynamic aspects of R-body extension.

R-bodies