Histoenzymological and ultrastructural changes in lateral muscle fibers of Oreochromis niloticus (Teleostei: Cichlidae) after local injection of veratrine

The effects of veratrine have been investigated in mammalian, amphibian, and crustacean muscle, but not in fish. In this work, the action of veratrine was studied in the lateral muscle of the freshwater teleost Oreochromis niloticus after intramuscular injection. Histoenzymological typing and electron microscopy of muscle fibers before and 15, 30, and 60 min after veratrine injection (10 ng/kg fish) were used to indirectly assess the morphological changes and the oxidative and m-ATPase activities. In some cases, muscles were pretreated with tetrodotoxin to determine whether the ultrastructural changes were the result of Na+ channel activation by veratrine. Veratrine altered the metabolism of fibers mainly after 30 min. Oxidative fibers showed decreased NADH-TR activity, whereas that of glycolytic and oxidative-glycolytic type fibers increased. There was no change in the m-ATPase activity of the three fiber types, except at 60 min postveratrine, when a novel fiber type, which showed no reversal after acidic and alkaline preincubations, appeared. Ultrastructural damage involved sarcomeres, myofibrils, and mitochondria, but the T-tubules remained intact. Pretreatment with tetrodotoxin (1 ng/ml) prevented the ultrastructural changes caused by veratrine. These results show that in fish skeletal muscle veratrine produces some effects that are not seen in mammalian muscle.

Distribution of acid phosphatases in the hypopharyngeal glands from workers, queens and males of a Brazilian stingless bee Scaptotrigona postica Latreille: An ultrastructural cytochemical study

The present study reports the localization of acid phosphatase in the hypopharyngeal gland cells from workers (newly-emerged, nurse and forager), queens (newly-emerged and laying) and males (newly-emerged and mature for mating) of the Brazilian stingless bee, Scaptotrigona postica. The phosphatase activity varied in intensity and localization depending on the individual class, physiological age and the substrate used. In newly-emerged workers, the phosphatase-positive sites suggest the involvement of the enzyme with cellular differentiation that occurs in the presecretory phase, in nurse workers with protein synthesis and in forager workers with changes in cellular activity or glandular regression. In males mature for mating and laying queens, the positive sites are related to secretory activity, showing that the gland maintains some activity in spite of the regressive aspect. Of the substrates used, β-glycerophosphate gave the least specific localization.

Submicroscopical features of leaves of xyris species

The leaf infrastructure of five Xyris species were examined using scanning electron microscope (SEM), transmission electron microscope (TEM) and histochemical methods. All studied leaves show some features in epidermis and mesophyll, which were of considerable adaptative significance to drought stress. Such features included the occurrence of a pectic layer on the stomatal guard cells and the presence of a nehvork ofpectic compounds in the cuticle. Pectic compunds were also in abundance in lamellated walls of the mesophyll cells and on the inner surface of the sclerified cell walls of the vascular bundle sheaths. There were also specialized chlorenchymatous peg cells in the mesophyll and drops of phenolic compounds inside the epidermal cells.

Calcium salts deposition in rat connective tissue after the implantation of calcium hydroxide-containing sealers

This study was conducted to observe the rat subcutaneous connective tissue reaction to implanted dentin tubes that were filled with mineral trioxide aggregate, Sealapex, Calciobiotic Root Canal Sealer (CRCS), Sealer 26, and the experimental material, Sealer Plus. The animals were sacrificed after 7 and 30 days, and the specimens were prepared for histological analysis after serial sections with a hard-tissue microtome. The undecalcified sections were examined with polarized light after staining according to the Von Kossa technique for calcium. At the tube openings, there were Von Kossa-positive granules that were birefringent to polarized light. Next to these granulations, there was irregular tissue, like a bridge, that was Von Kossa-positive. The dentin walls of the tubes exhibited a structure highly birefringent to polarized light, usually like a layer, in the tubules. These results were observed with all the studied materials, except the CRCS, which didn't exhibit any kind of mineralized structure. The results suggest that among the materials studied, the CRCS could have the least possibility of encouraging hard tissue deposition.

The ultrastructural aspects of vitellogenesis or oocyte secondary growth in Serrasalmus spilopleura (Teleostei, Characiformes, Serrasalminae)

Oocyte secondary growth in S. spiloleura corresponds to the period in which different vesicular structures are formed, including the cortical alveoli and the yolk granules. The oocytes with cortical alveolus formation show vesicular structures with filamentous content in the cortical cytoplasmic region, which are the cortical alveolus precursors. In these oocytes, electron-dense vesicles of heterogenous content are dispersed in the inner cytoplasmic region and their nuclei are irregular, showing many nucleoli of different sizes. The oocytes in vitellogenesis are filled with many vesicles. The cortical alveolus precursors are in the peripheral region, and electron-dense granules are seen near to the nucleus. These fuse and form yolk granules. The oocytes in vitellogenesis show a very irregular nucleus that has nucleoli of different sizes. In the oocytes in final vitellogenesis, the yolk granules are scattered throughout the cytoplasm, displacing the cortical alveoli toward cell periphery. The nucleus is similar to the other stages.

Kinetics of spermatogenesis in the Mongolian gerbil (Meriones unguiculatus)

The Mongolian gerbil (Meriones unguiculatus) is a small rodent native to the arid regions of Mongolia and Northeastern China. The present study provides descriptions of both the cellular associations of the seminiferous-epithelium cycle and relative frequencies of stages in the gerbil. Based on the development of the acrosomic system and the nuclear morphology changes using the PAS-H staining technique, the transformation of spermatids into spermatozoa was divided into 15 steps. The first 12 steps were used to identify 12 stages or cellular associations and the other three steps were spread among the first six stages of the cycle of the seminiferous epithelium. The relative frequencies found for stages I through XII were: 13.15; 8.06; 8.98; 6.48; 5.37; 6.71; 7.36; 7.45; 7.27; 5.83; 11.53 and 11.81, respectively. Stage I had the highest frequency while stage V proved the lowest frequency among the XII stages. The pattern of spermatogenesis is similar to those of rodents used as laboratory animals. The present description is the first for this rodent and provides the foundation for a variety of future studies of the testis in this animal. © 2002 Elsevier Science Ltd. All rights reserved.

Morphology and histoenzymology of eosinophilic granulocytes in the circulating blood of the turtle (Chrysemys dorbignih)

The studies on the characterization of eosinophils and neutrophils/ heterophils of turtles are contradictory. Some authors have pointed out the existence of two distinct cell types: eosinophils and heterophils. Other authors have proposed that eosinophils and heterophils may be the same cells in different stages of maturation. These interpretations are based only on a morphological analysis. In the blood of the turtle (Chrysemys dorbignih), a South American freshwater species, there are two types of granulocytes with eosinophilic staining pattern: the first with round cytoplasmic granules and the second with ellipsoidal cytoplasmic granules. In the present study by using histoenzymological methods for the analyses of enzymological cellular content, we found that the cells with round cytoplasmic granules were positive for nonspecific esterase and the cells with ellipsoidal granules were positives for acid phosphatase, alkaline phosphatase, nonspecific esterase and peroxidase. The results show that these cells are distinct cells and that the cells with ellipsoidal cytoplasmic granules have the same histoenzymological characteristics as the neutrophils/heterophils of mammalians and other vertebrates.

Mouse extensor digitorum longus and soleus show distinctive ultrastructural changes induced by veratrine

We investigated whether veratrine (5 μl, 10 ng/kg) injected into the mouse extensor digitorum longus (EDL) (fast-twitch) and soleus (SOL) (slow-twitch) muscles provokes distinctive ultrastructural disturbances 15, 30 and 60 min later. The mitochondria in SOL were affected earlier (within 15 min) than in EDL. Swelling of the sarcoplasmic reticulum terminal cisternae was more marked in EDL than in SOL and caused distortion of sarcomeres so that fragmentation of myofilaments was more pronounced in EDL. Hypercontracted sarcomeres were seen mainly in SOL and veratrine caused infoldings of the sarcolemma only in this muscle. In both muscles, the T-tubules remained unaffected and by 60 min after veratrine most of the above alterations had reverted to normal. Pretreatment with tetrodotoxin prevented the alterations induced by veratrine. This suggests that most of the alterations resulted from the enhanced influx of Na+ into muscle fibers. These results emphasize the importance of considering the type of muscle when studying the action of myotoxic agents.

The thrombocyte aggregation process in the turtle Phrynopys hilarii (Chelonia). An ultrastructural study

This study investigates the thrombocyte aggregation process in the South American fresh water turtle (Phrynopys hilarii) using electron microscopy. Blood was taken from surgically exposed lateral neck vessels often turtles Phrynopys hilarii during the spring and summer seasons, when the mean temperature is 37°C. Blood samples were fixed with Karnovsky solution for processing by transmission electron microscopy. The turtle thrombocytes were spindle-shaped with lobulated nuclei. Prominent vesicles and canaliculi were found throughout the cytoplasm. The cytoplasm organelles showed an agranular endoplasmatic reticulum, Golgi complex near the centrioles and scattered free ribosomes. These cells are similar to bird thrombocytes but distinct from fish and frog thrombocytes. Blood clotting time was 5 min ± 30 sec measured by the Lee and White method. Structural alterations resulting from the aggregation process occurred after activation. Thrombocytes developed numerous filopodial projections, an increased number of vacuoles and changed from spindle to spherical shape. P. hilarii thrombocytes have different morphologic characteristics compared to other non-mammalian vertebrate cells. These cells can participate in the aggregation process, as observed in birds.

Localization of peroxidase activity in blood mononuclear phagocytes in pacu fish (Piaractus mesopotamicus)

The localization of peroxidase activity in different cell regions is used as a criterion for classifying the stage of maturity of mammalian mononuclear phagocytes, with a positive peroxidase reaction indicating the presence of monoblasts, promonocytes, monocytes, and macrophages. Peroxidase activity was observed ultrastructurally in the circulating blood of pacu fish (Piaractus mesopotamicus), identifying monoblasts, promonocytes, monocytes, and macrophages. These observations suggest that differentiation of mononuclear phagocytes occurs in the blood circulation of fish, whereas in mammals, monoblasts and promonocytes are detected in bone marrow, with only monocytes detected in circulating blood and differentiation into macrophages occurring in other body compartments.

Cariostatic effect of fluoride-containing restorative systems associated with dentifrices on root dentin

Objective. The aim of this in vitro study was to evaluate the interaction between two sources of fluoride (restorative systems and dentifrices) in inhibiting artificial root caries development. Methods. One hundred and eighty tooth segments were embedded in polyester resin, and sanded flat. Cylindrical cavities 1.0 mm-deep and 1.5 mm-diameter were prepared in root dentin and randomly restored by fluoride-containing restorative systems: Ketac-fil/Espe (Ke), Fuji II LC/GC Corp (Fj), F2000/3M (F2), Surefil/Dentsply (Su) or a control: Filtek Z250/3M (Z2). Ten experimental groups were made to test the association among the five restorative systems and two dentifrices: with F - (Sensodyne Baking Soda) or without F- (Sensodyne Original) (n = 18). After surface polishing, a 1 mm-wide margin around the restorations was demarcated and initial dentin surface Knoop microhardness values (KHNi) were obtained. The specimens were submitted to a pH-cycling model, and to applications of slurries of dentifrice. Afterwards the final dentin surface Knoop microhardness values (KHNf) were measured. Results. The differences between KHNi and KHNf, and the covariate KHNi were considered by the ANCOVA and Tukey's test (α = 0.05). The interaction between restorative system and dentifrice was statistically significant (p = 0.0026). All restorative systems provided some protection against artificial caries challenge when associated with the fluoride-containing dentifrice treatment. The means (standard deviation) of reductions in Knoop hardness values for systems associated with the fluoride-containing dentifrice were: Ke: 40.0(1.02)a, Fj: 41.9(1.02)b, F2: 43.3(1.04)c, Su: 43.5(1.00)c, Z2: 44.0(1.02)c; and with the non-fluoride-containing dentifrice were: Ke: 42.9(1.02)a, Fj: 44.7(1.01)b, F2: 45.2(1.09)bc, Su: 46.0(0.99)c, Z2: 46.6(0.99)c (statistical differences were expressed by different letters). Conclusion. The cariostatic effect shown by the fluoride-containing dentifrice could enhance that shown by Ketac-fil and Fuji II LC, and could mask that shown by F2000. © 2002 Elsevier Science Ltd. All rights reserved.

Mating influence in the ovary differentiation in adult queens of Apis mellifera L. (Hymenoptera, Apidae)

The present results show that in the ovarioles of a newly emerged (0 day) queen of A. mellifera only two regions may be distinguished: a proximal, short germarium and a very long distal, terminal filament. As the queen matures and gets ready for the nupcial flight, the germarium increases in lenght, advancing towered the distal end, as the terminal filament shortens. The ovarioles of queens ready to mate (6 to 8 days old) have, already one or two ovarian follicles, i.e. a very short proximal vitellarium, but a real vitellogenesis only starts after the fecundation. If the queen does not mate the ovarioles structure is disrupted (12-16 days old). In mated queen eggs the ovarioles present three differentiated regions, from the apice to the basis: a short terminal filament, a medium size germarium, and a very long basal vitellarium. As the eggs are laid, the emptied follicle collapses, degenerates and produces a corpus luteum.

Spermiophagy in the spermatheca of Melipona bicolor lepeletier, 1836 (Hymenoptera, Apidae, Meliponini)

The presence of spermatozoa in vesicles in the cytoplasm of the epithelial cells that constitute the spermathecal wall of Melipona bicolor queen (Meliponini) is discussed in relation to the organ structure. The epithelial wall is lined by an apparently continuous cuticle in the luminal surface that should be a non-transposable barrier to the luminal spermatozoa. However, some spermatozoa were seen crossing the cuticle through interruptions that was first interpreted as sectioning defects. Nevertheless, the sperm cells in well-structured cytoplasmic vesicles, bound by membranes and sometimes associated to multivesicular bodies, as well as cytoplasmic structures representative of intracellular digestion and the occurrence of the phenomenon in two of the three spermathecae studied, suggest a real spermiophagic hole in the spermathecal epithelial cells.

Cytokeratins in epithelia of odontogenic neoplasms

Neoplasms and tumours related to the odontogenic apparatus may be composed only of epithelial tissue or epithelial tissue associated with odontogenic ectomesenchyme. The immunohistochemical detection of different cytokeratins (CKs) polypeptides and vimentin has made it easier to explain the histogenesis of many epithelial diseases. The present study aimed to describe the immunohistochemical expression of cytokeratins 7, 8, 10, 13, 14, 18, 19 and vimentin in the epithelial components of the dental germ and of five types of odontogenic tumours. The results were compared and histogenesis discussed. All cells of the dental germ were positive for CK14, except for the preameloblasts and secreting ameloblasts, in which CK14 was gradually replaced by CK19. CK7 was especially expressed in the cells of the Hertwig root sheath and the stellate reticulum. The dental lamina was the only structure to express CK13. The reduced epithelium of the enamel organ contained CK14 and occasionally CK13. Cells similar to the stellate reticulum, present in the ameloblastoma and in the ameloblastic fibroma, were positive for CK13, which indicates a nature other than that of the stellate reticulum of the normal dental germ. The expression of CK14 and the ultrastructural aspects of the adenomatoid odontogenic tumour probably indicated its origin in the reduced dental epithelium. Calcifying odontogenic epithelial tumour is thought to be composed of primordial cells due to the expression of vimentin. Odontomas exhibited an immunohistochemical profile similar to that of the dental germ. In conclusion, the typical IF of odontogenic epithelium was CK14, while CK8, 10 and 18 were absent. Cytokeratins 13 and 19 labelled squamous differentiation or epithelial cells near the surface epithelium, and CK7 had variable expression.