Sulphur-oxidizing extracellular bacteria in the gills of Mytilidae associated with wood falls

Six morphotypes of small mussels (Bivalvia: Mytilidae) were found attached to naturally sunken wood collected in the Bohol Sea (Philippines). These specimens are related to the large Bathymodiolus mussels that are found worldwide at cold seeps and hydrothermal vents. In these habitats, the mytilids harbour sulphur- and methane-oxidizing endosymbionts in their gills and depend on the energy and carbon provided by the symbionts. In this study, bacteria associated with the gills of wood-associated mussels are characterized using molecular and microscopic techniques. The existence of bacteria in the lateral zone of gill filaments in all specimens is demonstrated. Comparative analyses of 16S rRNA gene and adenosine 5'-phosphosulphate (APS) reductase gene sequences indicate that the bacteria are closely related to sulphur-oxidizing endosymbionts of Bathymodiolus. FISHs using specific probes confirm that sulphur oxidizers are by far the most abundant, if not the only bacteria present. Electron micrographs displayed mostly extracellular bacteria located between microvilli at the apical surface of host gill epithelial cells all along the lateral zone of each gill filament. In some specimens, occasional occurrence of intracellular bacteria with similar morphology was noted. This study provides the first molecular evidence for the presence of possible thiotrophic symbiosis in sunken wood ecosystems. With their epibiotic bacteria, wood-associated mussels display a less integrated type of interaction than described in their seep, vent and whale fall relatives.     

Three-dimensional reconstruction and quantitative analysis of rat lung type II cells: a computer-based study

The three-dimensional structure of alveolar epithelial type II cells was imaged using a computer-based system designed for reconstruction and quantitative analysis of serially sectioned specimens. Six type II cells were reconstructed from serial ultrathin sections of lungs from two Sprague Dawley male rats and the results were compared to standard morphometric estimates of type II cell composition from five other Sprague Dawley male rats. A minor portion of the type II cell surface was in contact with the alveolar airspace while most of the cell surface was embedded in the alveolar septal interstitium. The type II cells contained multiple Golgi regions located close to the nucleus. Mitochondria formed a few branching filamentous networks extending throughout the cell. The reconstructed cells appeared to represent a homogeneous population having fractional volumes of intracellular organelles very similar to those found by morphometric techniques. The spatial distribution of secretory organelle volume suggests that the organization of this cell type reflects an ordered progression of secretory particle maturation which is consistent with earlier hypotheses of lamellar body assembly.     

热牙胶充填与冷牙胶侧方加压在根管充填中应用的临床效果比较

目的:比较热牙胶充填与冷牙胶侧方加压在根管充填中应用的临床效果。方法:选择62例急性牙髓炎和根尖周炎患者,均分后分别选择热牙胶充填与冷牙胶侧方加压的方法进行根管充填,比较两组患者根管充填时间、术后牙周疼痛病例和术后3个月后充填质量及临床疗效。结果:两组患者术后牙周疼痛病例比较,差异无统计学意义(P>0.05);其余各方面比较,差异均均有统计学意义(P>0.05),热牙胶组优于冷牙胶组。结论:临床进行根管充填治疗时,应首选热牙胶充填的方式,可显著提高临床疗效,且不增加患者术后牙周疼痛的发生。     

Apical and basal Forssman antigen in MDCK II cells: a morphological and quantitative study.

We have studied the surface distribution of a glycosphingolipid (the Forssman antigen) in MDCK II and CCL39 cells. The Forssman antigen is mobile on the surface of both these cell lines. Its surface distribution is homogenous on non-polarized cells. Under conditions where MDCK II cells are well polarized, the Forssman antigen is present in equal amounts on the apical membrane and on the basal membrane and its processes. Very little Forssman antigen can be detected on the lateral membrane. The nature of the mechanism excluding the Forssman antigen from the lateral domain remains to be determined. This surface distribution is established within hours after plating and was observed with cells grown on different types of filters. The surface density of the Forssman antigen on the apical and on the basal domain has been estimated. No involvement of the basal Forssman antigen in cell attachment could be demonstrated. However, the apical Forssman antigen appears to be essential to the establishment of the cells in culture.     

THE FINE STRUCTURE OF VON EBNER''S GLAND OF THE RAT

The fine structure of von Ebner's gland was studied in untreated rats and rats stimulated to secrete by fasting-refeeding or injection of pilocarpine. Cytological features were similar to those reported for pancreas and parotid gland. Abundant granular endoplasmic reticulum filled the basal portion of the cell, a well-developed Golgi complex was located in the vicinity of the nucleus, and the apical portion of the cell was filled with dense secretory granules. Dense heterogeneous bodies resembling lysosomes were closely associated with the Golgi complex. Coated vesicles were seen in the Golgi region and also in continuity with the cell membrane. Granule discharge occurred by fusion of the granule membrane with the cell membrane at the secretory surface. Successive fusion of adjacent granules to the previously fused granule formed a connected string of granules in the apical cytoplasm. Myoepithelial cells were present within the basement membrane, and nerve processes were seen adjacent to acinar and myoepithelial cells. Duct cells resembled the intercalated duct cells of the major salivary glands.     

Anatomical aspects of the gynoecium of species of Anthurium sect. Urospadix Engl. (Araceae,Alismatales)

Anthurium sect. Urospadix includes approximately 70 species differently distributed in Eastern Brazil, Western South America, and Central America. The present work studied the gynoecia of species of section Urospadix to describe their anatomical features. In all the species studied, the apical portion of the gynoecium differentiates into a stigma covered with short trichomes and an adjacent region formed by isodiametric cells. The stylar canal comprises short, unicellular secretory trichomes extending up to the ovarian aperture. Carpels are apically symplicate and basally synascidiate with a septum that does not reach the apical portion of the locule. The apical portion of the septum differentiates into long, secretory trichomes while the subapical one differentiates into the placenta. Locules are filled with mucilage. Since the epidermis of the region adjacent to the stigma, the length of the stylar portion, and the number of cell layers of the ovarian septum vary among the species studied, they may be useful for further taxonomic and systematic purposes. The present work is the first detailed report of trichome location and distribution on the gynoecia of Anthurium.     

The Effect of Chilling of the Physiological and Simulated Apex of 2- and 3-leaf Plants of Pisum sativum L. cv. Alaska on Lateral Shoot Growth, C-14 IAA Movement and P-32 Accumulation

The apex of a 3-leaf pea plant was chilled in cold chambers maintained at 5–7°C. The lateral shoots 1 through 5 grew, and shoot 5 eventually dominated other lateral shoots. The apex when returned to the ambient temperature did not reimpose apical dominance. The growing lateral shoots competed with the stem apex. The apices of 2- and 3-leaf plants were chilled and P-32 distribution in these plants was studied in the entire plant, at various intervals of time. Phosphorus-32 accumulation followed the growth pattern of the plant. The lateral shoots accumulated P-32 activity and very little activity was accumulated by the apex. The dominating shoots 2 and 5 accumulated the maximum amount of activity in 2- and 3-leaf plants, respectively. Labeled-IAA moved basipetally through the stem when applied to the cut stump simulating the apex. By cold treatment the translocation of IAA was influenced more than its absorption. The plant seems to metabolize this compound in the later periods of application. The plant now becomes “insensitive” to auxin and the lateral shoots grow.     

Key differences between lateral and apical branching in hyphae of Neurospora crassa

We examined in fine detail growth kinetics and intracellular events during lateral and apical branching in hyphae of Neurospora crassa. By high-resolution video-enhanced light microscopy, we found remarkable differences in the events preceding lateral vs apical branching. While apical branching involved a significant disturbance in the apical growth of the parental hypha, lateral branching occurred without any detectable alterations in the growth of the parental hypha. Prior to the emergence of a lateral branch, an incipient Spitzenk?rper was formed about 12-29 microm behind the apex. Lateral branch formation did not interfere with the elongation rate of the primary hypha, the shape of its apex or the behavior of its Spitzenk?rper. In sharp contrast, apical branching was preceded by marked changes in physiology and morphology of the parental hypha and by a sharp drop in elongation rate. The sequence involved a cytoplasmic contraction, followed by a retraction, dislocation, and disappearance of the Spitzenk?rper; hyphal elongation decreased sharply and a transient phase of isotropic growth caused the hyphal apex to round up. Growth resumed with the formation of two or more apical branches, each one with a Spitzenk?rper formed by gradual condensation of phase-dark material (vesicles) around an invisible nucleation site. The observed dissimilarities between lateral and apical branching suggest that these morphogenetic pathways are triggered differently. Whereas apical branching may be traced to a sudden discrete disruption in cytoplasmic organization (cytoplasmic contraction), the trigger of lateral branching probably stems from the subapical accumulation of wall precursors (presumably vesicles) reaching a critical concentration.     

Morphological aspects of Culex quinquefasciatus salivary glands

The salivary glands of Culex quinquefasciatus female mosquitoes are paired organs composed of two lateral lobes with proximal and distal secretory portions, and a medial lobe. All portions comprise a simple epithelium that surrounds a salivary duct. In the apical portion of the medial lobe, non-secretory cells strongly resemble cells involved in ion and water transport. The general architecture of the secretory portions is similar between lobes. The appearance of the secretory material and the morphological aspect of the apical cell membrane are the most distinctive features among the three secretory portions. Cells in the lateral proximal lobe display thin membrane projections extending into a translucent and finely filamentous secretory product. At the lateral distal portion, the apical cell membrane forms an intricate meshwork that encloses a dark secretory product. Medial lobe secretory cells also contain secretory cavities surrounded by intracytoplasmic vesicles, all containing a very dark and uniform product. Scattered cells holding numerous vacuoles, some of them containing a small and electron-dense granule eccentrically located and resembling those of the diffuse endocrine system, are frequently observed in the periphery of all secretory portions. Immunofluorescence assays revealed that the distal portion of the lateral lobes contains apyrase, an enzyme putatively responsible for platelet aggregation inhibition, diffusely distributed in the cell cytoplasm.     

A Two-Dimensional Vibrating Probe Study of Currents around Lateral Roots of Raphanus sativus Developing in Culture

A computer-assisted, two-dimensional vibrating probe was used to study the ionic currents around developing lateral roots of Raphanus sativus in vitro. This system allowed us to superimpose current vectors on the video image of the roots. In a young lateral root, current entered the cap, meristematic, and elongation zones and exited the primary root surface close to the base of the lateral root. As the lateral root grew, current began to exit from its basal (cell maturation zone) end. The densities of currents entering the apical portion of the faster-growing lateral roots in a medium lacking indole 3-acetic acid were about twice as large as those entering the apical region of the slower-growing lateral roots in indole 3-acetic acid-supplemented medium.     

Apoptosis, cell proliferation and serotonin immunoreactivity in gut of Liza aurata from natural heavy metal polluted environments: preliminary observations

In the present paper, the effect of natural environment non-lethal heavy metal concentration on cell renewal of Liza aurata intestinal epithelium, was studied by the TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling) method and anti-PCNA (proliferating cell nuclear antigen) immunohistochemistry, in order to detect, respectively, apoptosis and cell proliferation. In addition, the presence and distribution of the cell renewal regulator, serotonin, was immunohistochemically investigated. In order to reduce variability, only immature specimens were considered. The results indicated that in the control specimens from non-polluted areas, the PCNA immunoreactive nuclei of the proximal intestinal epithelium were only located at the bottom of the intestinal folds, together with a few TUNEL-positive nuclei, and goblet mucous differentiated cells. In the specimens from polluted areas, the number of PCNA immunoreactive cells was greatly enhanced, and they extended along the mid portion of the intestinal folds; the number of TUNEL-positive nuclei was enhanced as well, but they were almost exclusively detected in the third apical portion of the intestinal folds. Serotonin immunoreactive nerve elements were more frequently detected in the intestinal wall of L. aurata specimens from polluted areas, and besides that, some serotonin immunoreactive endocrine cells were also present. Variations in distribution and frequency of TUNEL-positive nuclei, PCNA immunoreactive nuclei, and serotonin immunoreactivity put in evidence an alteration of cell renewal with an enhancement of cell proliferation, probably leading to morphological intestinal fold changes.     

Changes of endogenous hormones in lateral buds of chrysanthemum during their outgrowth

The character of branching for two chrysanthemum (Chrysanthemum × morifolium) cvs. Jinghai and Jingyun was observed, and the changes of endogenous hormones in apical and lateral buds were investigated to determine the relationship between the pattern of hormone distribution, apical dominance, and lateral bud outgrowth. The growth rate of Jinghai lateral buds was higher than that of Jingyun. In vegetative growth stage, IAA level in apical buds of Jingyun was significantly higher than in Jinghai. After flower induction, IAA level in apical buds of two cultivars decreased remarkably, but the IAA level decreased in Jingyun faster than in Jinghai. These results showed that the higher was the IAA level in apical buds the stronger was inhibition of lateral bud outgrowth. An increase in IAA and iP/iPA and a decrease in ABA concentrations were closely associated with lateral bud growth alterations in chrysanthemum.     

Surface changes in denture soft liners with and without sealer coating following abrasion with mechanical brushing

Gerodontology 2010; doi: 10.1111/j.1741‐2358.2010.00375.x Surface changes in denture soft liners with and without sealer coating following abrasion with mechanical brushing Aim: To evaluate the surface alterations of soft liners with or without sealer coating following abrasion with mechanical brushing. Methods: Thirty specimens were made of a methacrylate‐ (Coe‐Soft) and a siloxane‐based material (Ufi‐Gel SC), and 15 received two coatings of surface sealer. The specimens were submitted to a mechanical brushing‐dentifrice assay under 200 g of force at 250 cycles/min. Mechanical brushing was simulated for a period of 1 (1250 cycles) and 6 months (5000 cycles). Surface roughness (Ra parameter) was measured, and scanning electron microscopy (SEM) images were obtained. Ra data were analysed by anova for repeated measures and Bonferroni’s test (alpha = 0.05). Results: Ra increased from baseline to 6 months regardless of sealer coating. At baseline, only Coe‐Soft without sealer had a higher Ra than the other groups. After 1 month, the Ra of Coe‐Soft with sealer was three‐fold higher than the Ra at baseline; the other groups showed no significant increase of Ra. SEM images showed degradation of the soft liners over time, except for the Ufi‐Gel SC with sealer, which displayed minimum alteration of surface texture. Conclusion: Sealer coating reduced the surface degradation of the tested soft liners, but the protective effect was more pronounced for the siloxane‐based material.     

Immunohistochemical localization of NPY, VIP and 5-HT in the thyroid gland of the lizard, Podarcis sicula

The thyroid gland of the lizard Podarcis sicula was immunohistochemically studied in adult male specimens using specific antibodies against NPY, VIP and 5-HT and the avidin-biotin peroxidase complex (ABC) procedure to localize the three peptides. Fine beaded VIP-immunoreactive nerve fibers ran between the follicles, and VIP-immunoreactivity was evenly distributed in the apical cytoplasm of follicular cells. NPY-immunoreactive fibers were found around the follicles, and, in the cells, immunoreactivity was localizated only in the cellular apices. Immunoreactivity to 5-HT was observed in the colloid, with a concentration in the follicular lumen exceeding that in the follicular cells. In fact, most follicles showed immunoreactivity in the cytoplasmic bridges formed between the apical portion of the follicular cells and the colloid.     

Extant silicoflagellates from the Northeast Aegean (eastern Mediterranean Sea): Morphologies and double skeletons

Silicoflagellate abundance, vertical distribution and morphology were studied during spring (March 2014) at three sampling stations located in the Northeast Aegean Sea adjacent to the Dardanelles Strait and characterized by a variable influx of cold, low-salinity Black Sea water. The silicoflagellate assemblage was dominated by Dictyocha stapedia and Stephanocha speculum with minor contribution of D. aculeata and Octactis pulchra. While specimens of D. stapedia were represented by the typical morphologies described in other areas of the Mediterranean Sea, populations of S. speculum displayed peculiar characters: they were large, predominantly 7-sided, with a small apical ring as well as apical ring spines, concave basal ring sides and non-rotated apical structure. Some of these features have been described for S. speculum at high latitudes, but the combined characters make these specimens slightly different from the high latitude populations. Similar morphologies have been observed in the western Black Sea, thus we can infer that the peculiar specimens detected in the Northeast Aegean are associated with the influx of Black Sea water masses.     

Hierarchy of mechanisms involved in generating Na/K-ATPase polarity in MDCK epithelial cells

We have studied mechanisms involved in generating a polarized distribution of Na/K-ATPase in the basal-lateral membrane of two clones of MDCK II cells. Both clones exhibit polarized distributions of marker proteins of the apical and basal-lateral membranes, including Na/K- ATPase, at steady state. Newly synthesized Na/K-ATPase, however, is delivered from the Golgi complex to both apical and basal-lateral membranes of one clone (II/J), and to the basal-lateral membrane of the other clone (II/G); Na/K-ATPase is selectively retained in the basal- lateral membrane resulting in the generation of complete cell surface polarity in both clones. Another basal-lateral membrane protein, E- cadherin, is sorted to the basal-lateral membrane in both MDCK clones, demonstrating that there is not a general sorting defect for basal- lateral membrane proteins in clone II/J cells. A glycosyl- phosphatidylinositol (GPI)-anchored protein (GP-2) and a glycosphingolipid (glucosylceramide, GlcCer) are preferentially transported to the apical membrane in clone II/G cells, but, in clone II/J cells, GP-2 and GlcCer are delivered equally to both apical and basal-lateral membranes, similar to Na/K-ATPase. To examine this apparent inter-relationship between sorting of GlcCer, GP-2 and Na/K- ATPase, sphingolipid synthesis was inhibited in clone II/G cells with the fungal metabolite, Fumonisin B1 (FB1). In the presence of FB1, GP-2 and Na/K-ATPase are delivered to both apical and basal-lateral membranes, similar to clone II/J cells; FB1 had no effect on sorting of E-cadherin to the basal-lateral membrane of II/G cells. Addition of exogenous ceramide, to circumvent the FB1 block, restored GP-2 and Na/K- ATPase sorting to the apical and basal-lateral membranes, respectively. These results show that the generation of complete cell surface polarity of Na/K-ATPase involves a hierarchy of sorting mechanisms in the Golgi complex and plasma membrane, and that Na/K-ATPase sorting in the Golgi complex of MDCK cells may be regulated by exclusion from an apical pathway(s). These results also provide new insights into sorting pathways for other apical and basal-lateral membrane proteins.     

Ultrastructural and immunohistochemical studies on non-ciliated cells of the tracheal epithelium of normal,phenobarbital-treated,and 3-methylcholanthrene-treated mice

Summary Non-ciliated SER-rich cells of the tracheal epithelium of normal, phenobarbital-treated and 3-methylcholanthrene-treated mice were studied ultrastructurally and immunohistochemically. The apical portion of these cells protrudes into the tracheal lumen, especially in the mice treated with the two compounds, and the apical cytoplasm is filled with numerous tubular elements of SER. Besides, the non-ciliated cells of 3-methylcholanthrene-treated mice show a strong positive reaction to the antiserum against microsomal cytochrome P-450 of liver. These findings support the concept that the non-ciliated tracheal cell may be involved in the metabolism of endogeneous and exogeneous chemical compounds.This study was supported by grants from the Ministry of Education, Science and Culture, Japan     

Regulatory peptides in the urinary bladder of two genera of Antarctic teleosts

Somatostatin 14, prolactin, atrial natriuretic peptide, galanin and urotensin II were found using immunohistochemistry in the urinary bladders of the Antarctic fishes Trematomus bernacchii (Nototheniidae) and Chionodraco hamatus (Channichthyidae) caught in the Ross Sea. The urinary bladders of the two species showed a different histology in the epithelial layer. In T. bernacchii the epithelium comprises a single type of columnar cells, while in C. hamatus the columnar cells are restricted to the ventral portion of the bladder, and the dorso-lateral region is lined by cuboidal cells. No difference in the intensity of the immunostaining was observed in the two cell types; the only variation was a different distribution of the immunoreactions, which were present in the whole cytoplasm in the cuboidal cells and restricted to the apical and/or basal portion of the columnar cells. Accepted: 1 November 1998     

Agrobacterium-mediated transformation of Pinus radiata organogenic tissue using vacuum-infiltration

An Agrobacterium tumefaciens-mediated transformation protocol was developed for detached cotyledons of Pinus radiata zygotic embryos resulting in up to 55% of cotyledons transiently expressing the reporter gene uidA. Transient expression of uidA was improved when detached cotyledons were pre-cultured on half strength medium containing cytokinin for 7 days, wounded by vortexing and then vacuum-infiltrated in a solution of A. tumefaciens. The transformation protocol was applied both to cotyledons and also to the apical meristematic dome which was the portion of the embryo remaining after cotyledons were detached, and from which the apical shoot and axillary shoots regenerate. Molecular analysis of putatively transformed shoots regenerated either adventitiously from cotyledons or via axillary shoots from apical domes, indicated the presence of uidA and nptII genes by PCR in some of these shoots. Biochemical analysis of putatively transformed shoots using nptII ELISA indicated that they contained the nptII enzyme. However, Southern hybridisation indicated stable integration of nptII only in one shoot which was regenerated from an apical dome. Shoots regenerated from cotyledons appeared to exhibit chimeric expression and were not stably transformed. Based on a comparison of time for regeneration, technical difficulty, molecular and biochemical analysis, apical domes may be more suitable as explants for transformation and subsequent regeneration of transclones than detached cotyledons.     

Structure of the urinary bladder in the Pacific cod

The urinary bladder of the Pacific cod,Gadus macrocephalus was investigated. A pair of ureters was found to unite and form a sac-like protrusion on the posteroventral aspect of the gonad. This protrusion was confirmed as the urinary bladder based on its histology and the presence of ammonia and urea in the fluid within this structure. The urinary bladder consisted of a central lumen and two lateral expansions found on the right and left sections. The luminal epithelial cells of the Pacific cod urinary bladder, unlike those of other animals, were characterized by microvilli on the free surface, and the presence of a number of vesicles and mitochondria across the apical portion on the cells. This suggests that the luminal epithelium of the urinary bladder might be actively engaged in transportation of water or other materials from the urine. The Pacific cod urinary bladder may therefore be closely associated with osmoregulation by reabsorbing water from the urine. Ammonia-N and urea-N levels of 65–213 μg/dl and 1.5–3.5 mg/dl were measured in the fluid which filled the urinary bladder.