Journal of submicroscopic cytology and pathology最新文献

The in vitro cytopathic effect of four strains of Trichomonas vaginalis on cultured epithelial monolayers was analyzed through electrophysiology and electron microscopy. Interaction of trichomonads of two virulent strains (GT-10 and GT-13) with cultured MDCK cell monolayers mounted in Ussing chambers produced a rapid decrease in transepithelial resistance to less than 30% of control values after only 15 min. By 30 min the electrical resistance was practically abolished by the virulent parasites. In contrast, of two attenuated strains of trichomonads (GT-3 and GT-7) analyzed under similar conditions, GT-3 trophozoites required 180 min to reduce transepithelial resistance to 9% of control values, while monolayers in contact with GT-7 parasites still showed 28% of control values at this time of incubation. Sequential scanning electron microscopy confirmed the much faster and widespread cytopathic effect of virulent parasites. In contrast, the slow lytic process produced by attenuated trophozoites was reduced to focal areas of direct contact with epithelial cells. Another difference was found by measurement of the surface charge of the four strains of T. vaginalis by means of cell microelectrophoresis. While the two virulent strains showed a negative surface charge, the two attenuated strains had no detectable surface charge at neutral pH. When parasites were incubated with cationized ferritin and studied with transmission electron microscopy the surface of virulent trichomonads appeared heavily labeled, whereas the surface of attenuated parasites had only sparse and irregular ferritin binding.

The spermatozoa of Exomalopsis auropilosa and Paratetrapedia (Lophopedia) sp. are long and slender, measuring about 374 microm and 370 microm in length of which the head region measures approximately 25.8 and 28.3 microm, respectively. The head consists of an acrosome formed by an acrosomal vesicle covering a perforatorium, which presents a paracrystalline organization in Paratetrapedia (Lophopedia) sp. and a nucleus. This latter measures about 24 microm in Exomalopsis auropilosa and 27 microm in Paratetrapedia (Lophopedia) sp., and has compact chromatin. The nucleus is attached to the flagellum by an electron dense material and centriolar adjunct is observed between it and smaller mitochondrial derivative. In this flagellar region only one accessory body is observed, which occurs between the larger mitochondrial derivative and the axoneme. The flagellum consists in a typical axoneme, 9+9+2 microtubules, two mitochondrial derivatives and two accessory bodies. The two mitochondrial derivatives are asymmetric in both length and diameter, and paracrystalline material appears only in the larger mitochondrial derivative. The structure and ultrastructure of the spermatozoa of the bee species here described are similar to the majority of sperm found in the other Hymenoptera and may be a contribution for future phylogenetic analysis of Apidae.

We performed a scanning electron microscopy study on the human urinary bladder tunica mucosa. Specimens from bladder biopsies were treated with OsO4 maceration and 1N NaOH maceration methods prior to SEM observation to disclose the three-dimensional organization of the lamina propria, basal lamina and urothelium. The lamina propria housed a well developed capillary plexus just below the basal lamina; the urothelium presented a typical three-layered organization with basal, intermediate and superficial cells. The intermediate cells appeared essentially similar to basal cells in their external features and stretched from the basal lamina up to the superficial layer. The most superficial cells appeared consistently flattened and interconnected by extensive junctional complexes. They showed a peculiar specialization, their apical plasmalemma being thickened with distinctive, stiff plaques, in contrast with the underlying globular or spindle-shaped cells whose plasmalemma was only covered by short microvillosities.

The gap junctions between perineuronal satellite cells were studied in the spinal ganglia of 12, 42, and 79-month-old rabbits. The mean number of gap junctions per 100 microm2 of surface of the section occupied by satellite cells was significantly greater in old rabbits than young adults. Since the mean length of individual gap junctions did not change with age, the increase in number of gap junctions cannot be due to fragmentation of pre-existing gap junctions but is very likely due to the formation of new gap junctions. The increase in number of gap junctions cannot be related to an increase in number of perineuronal satellite cells since the mean number of these cells is significantly smaller in aged rabbits than in young adults. It is suggested that the increase in number of gap junctions with age may enhance the suggested neuroprotective role of satellite cells towards ganglionic neurons. The present findings, together with previous observations, suggest that the gap junctions between perineuronal satellite cells are dynamic structures, able to adapt to varying neuronal demands and varying environmental conditions.

The effects of diets with variable zinc levels on the midgut epithelial cells were studied in Oreochromis niloticus L. One hundred and twenty fry of tilapia were apportioned into 4 experimental groups (I, II, III and IV groups), with 30 fish in each treatment, 5 replicate aquaria per treatment containing 6 fish each. The animals of the 4 groups were fed with isonitrogenous (30% crude protein) and isoenergetic (3000 Kcal/Kg of digestible energy) diets with increasing quantities of zinc (44.59; 149.17; 309.93; 599.67 mg Zn/kg of diet), twice a day, for 93 days. Three fish from each group were sacrificed at 36, 66 and 93 days and samples of midgut were removed for ultrastructural analysis. After 93 days of treatment, 3 animals of each experimental group were used for the analysis of zinc concentration by atomic absorption spectrophotometry. The comparative relative index (CRI) revealed that the animals in groups II, III and IV contained, respectively, 1.99%, 34.67% and 22.78% more zinc than the mean concentration in animals from group I. The ultrastructural analysis showed enterocytes with swelling of smooth surfaced endoplasmic reticulum and dilated mitochondria with variable matrix rarefaction and cristae number reduction in the fish exposed to 599.67 mg Zn/Kg of diet at 66 and 93 days of treatment.

Two neural circuits in the retina are described, that involve neural interaction generated by image movement. One circuit increases the sensitivity in detecting movement in the visual field while the other circuit generates ganglion cell signals when an image moves in one preferred direction but not when moves in the opposite direction, explaining the observation of directionally selective ganglion cell responses described by Barlow and Hill (1963). In agreement with the recorded potentials, the circuit generates ganglion cell responses also when the light stimulus is stationary. This ambiguity in ganglion cell signals to the visual center limits the signals usefulness to images that are in constant movement. The only known such movement is that caused by the involuntary eye movements. The function of the circuit is therefore to stabilize the retinal image when transmitted to the visual center.

The functional role played by test cells in larvae of various ascidian species consists in depositing sub-microscopic structures known as ornaments and/or proteoglycan substances on the larval test surface. According to the data reported in the literature, the deposition of ornaments together with proteoglycan substances on the larval test would render the latter hydrophilic and thus allow the larva to swim being immersed in water. Ornament deposition on the larval test does not occur in all the ascidian species. Ultrastructural investigations made on larvae belonging to the Cionidae and Ascididae families, for instance, have failed to evidence the presence of ornaments on the test. For these ascidian families it has been hypothesized that in swimming larvae test cells secrete an amorphous substance that would allow them to adhere to the larval test. In order to ascertain the functional role played by test cells in swimming larvae of the Ascididae family, the presently reported ultrastructural and cytochemical investigations have been made on larvae of Ascidia malaca. Besides suggesting that test cells, tightly adherent to the test surface, present an amoeboidic behaviour, the ultrastructural investigations have evidenced that these cells are still metabolically active. Their cytoplasm, characterized by the presence of a Golgi apparatus actively involved in synthesis, is almost entirely filled with very large granules; some of them gradually empty their contents turning into vacuoles containing scarce residues of electrondense particles. The present ultrastructural observations support the hypothesis that the adhesion of test cells on the larval test could be very likely eased by the secretion of substances synthesized by the Golgi and released through pseudopodes which test cells then wedge into the test. The cytochemical investigations were based on a reaction (fixation in glutaraldehyde-tannic acid) which evidences the presence, at the ultrastructural level, of proteoglycan substances such as glycosaminoglycans (Singley and Solursh, 1980). The reaction has given positive results in test cell granules undergoing emptying, on the outer membrane of the same cells, and on the outer cuticular layer C1 of the larval test. The present investigations, besides confirming the absence of ornament deposition on the test surface by test cells of Ascidia malaca swimming larvae, have evidenced that the secretion products deposited on the larval test surface by test cells consist of glycosaminoglycans, i.e. proteoglycan substances. In agreement with the data reported in the literature, it is hypothesized that the deposition of glycosaminoglycans on the surface of Ascidia malaca larval test makes the larval tunic hydrophilic and thus the larva is able to swim being immersed in water.

Clinical and biopsy study of nine patients on statin therapy suffering from various myopathic syndromes is reported. Biopsy findings showed non specific myopathic signs and mitochondrial changes, such as subsarcolemmal accumulation, morphological alterations, lipid increase and Cox-negative fibers. These findings confirm that statins may cause muscle damage and impair oxidative metabolism.

The Ag-NOR staining technique and image analysis were used to evaluate morphological parameters (area, perimeter and axis ratio) in nucleoli from normal thyroids and from thyroids bearing proliferating lesions (carcinomas, adenomas and hyperplasias). Regions with normal appearance located close to adenomatous and carcinomatous regions, in the thyroid of every patient, were also analyzed for comparison with the respective pathological regions and with normal thyroids. Statistical analysis of data for the nucleolar area and perimeter allowed the separation of adenomas and carcinomas from hyperplasias and normal tissue but not the two components in each of these two groups. However, if we look at the numbers, a sequence of increasing nucleolar mean areas in the order: normal, hyperplasia, adenoma and carcinoma may be observed, indicating the sequence of increasing rRNA requirements in these different kinds of cells. The axis ratio that denotes the nucleolar shape (round or oblong) did not show significant differences among tissues, suggesting that shape is not important in the characterization of these pathologies. Differences in nucleolar areas and perimeter between normal and affected regions from each patient were statistically significant for adenomas and carcinomas. When these normal regions were compared with the normal thyroids, significant differences were not obtained in the three evaluated parameters. The observations and their importance for histopathological diagnosis are discussed.

Biopsy specimens of cervico-scutular muscles obtained from animals injected with bee crude venom were prepared for electron microscopy studies. At 6 h from Apis mellifera venom injection, in mice under transmission electron microscopy, the muscular fibres presented different atrophy levels with increment of the intermyofibrillar spaces. Tubules and sarcoplasmic reticulum elements were altered, in some places only tubular fragments and an increment of the intermyofibrillar spaces were noticed as well as loss of fibre regularity and prominent triads. In subsarcolemma region, areas lacking myofibrils and mitochondria damages were observed. Muscular segmental necrosis and atrophy areas were observed. Neuromuscular junctions were altered. The number of synaptic vesicles was very variable and synaptic clefts showed irregularities. A decrease in the number and arrangement of the synaptic clefts, as well as free polysomes, suggesting regeneration processes, were also observed. The myelinic nerves exhibited in the axon or in the wall vacuolisation areas. The presence of severe muscular lesions, the finding of venom activities in the presynaptic region and the detection of damages in the neuromuscular junctions at different chronological stages of our experiments indicate that the bee venom is highly toxic for neuromuscular structures.