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Microscopic Analysis (microscopic + analysis)
Kinds of Microscopic Analysis Selected AbstractsLight and Electron Microscopic Analyses of Autologous Pericardial Tissue Used as a Small-Diameter Arterial Graft in DogsARTIFICIAL ORGANS, Issue 1 2002Won Gon Kim Abstract: As a form of small-diameter arterial graft, we implanted fresh autologous pericardium and pericardium treated with 0.6% glutaraldehyde in the bilateral carotid arteries of dogs and then compared the time-related changes of the grafts explanted after the predetermined periods. The pericardial grafts were implanted in 1 animal each for scheduled periods of 3 days, 2 weeks, 1 month, 3 months, and 6 months. The retrieved grafts were processed for light and electron microscopic analyses following gross observation. The glutaraldehyde-treated small-diameter pericardial vascular grafts showed a better endothelialization of the blood-contacting surface and a slower fragmentation of the collagen layers than the fresh grafts although it has yet to be proven whether these differences are so significant as to affect the patency results between the groups. [source] ,Defence lignin' and hydroxycinnamyl alcohol dehydrogenase activities in wounded Eucalyptus gunniiFOREST PATHOLOGY, Issue 2 2003S. Hawkins Summary To learn more about lignin formation in response to wounding in trees, we adopted two complementary approaches: (1) microscopic and histochemical studies of the wound response in 3.5-month-old Eucalyptus gunnii plantlets and (2) biochemical investigations of hydroxycinnamyl alcohol dehydrogenase activities in wounded 6-year-old, field-grown E. gunnii trees. The first approach revealed that a barrier zone was formed in response to wounding in both ground tissues (cortex barrier and pith reaction zone) and vascular tissues. The barrier zone was barely detectable after 24 h but well-developed 7 days after wounding. Microscopic analyses indicated that the barrier zone was formed by the reinforcement of cell walls with ,lignin-like material' in both ground tissues and vascular tissue, and that, in addition, the lumen of certain xylem cells (vessels and fibres) were blocked by the deposition of polymeric phenolic material. Histochemical characterization revealed that the lignin-like material (,defence lignin') deposited in ground tissue cell walls and xylem cell blockages was poor in syringyl (S-type) lignin units and therefore differed from the usual mixed guaiacyl,syringyl (G,S) lignin unit composition of E. gunnii developmental lignin. In contrast, S-type lignin appeared to be deposited in the cell walls of immature developing secondary xylem cells at a stage when the cell walls of comparable cells from unwounded control stems contained lignin poor in syringyl units. The second approach indicated that two different types of cinnamyl alcohol dehydrogenase activity are induced, and apparently regulated differentially, in response to wounding in E. gunnii trees. Coniferyl alcohol dehydrogenase activity was induced immediately and continued to increase throughout the first 15 days of the 17-day experimental period, while sinapyl alcohol dehydrogenase activity was first detected at 8 days after wounding and continued to increase throughout the experimental period. The biological roles of the two alcohol dehydrogenase activities are discussed in relation to the formation of defence lignin versus developmental lignin in trees. Résumé Afin d'approfondir nos connaissances concernant la formation de lignine en réponse aux blessures chez les arbres, nous avons utilisé deux approches complémentaires: (1) des études microscopiques et histochimiques de la réponse à la blessure chez les plantules d'Eucalyptus gunniiâgées de 3 mois et demi, et (2) des analyses biochimiques des activités alcools hydroxycinnamyliques déshydrogénases chez les arbres âgés de 6 ans blessés au champ. L'utilization de la première approche a révélé qu'une barrière physique se forme en réponse à la blessure aux niveaux des tissus vasculaires, de la moelle, et des tissus externes au phloème. A 24 h après la blessure, cette barrière est peu développée mais elle est bien formée après 7 jours. Les analyses microscopiques et histochimiques ont indiqué que les parois cellulaires au niveau de la barrière sont renforcées par un composé semblable à la lignine (,lignin-like material'). De plus, les lumens de plusieurs cellules xylémiennes (vaisseaux et fibres) sont bouchées par le dépôt d'un composé polymérique de nature phénolique. Les caractérizations histochimiques ont indiqué que le ,lignin-like material' (lignine de défense) déposé dans les parois cellulaires de la moelle et des tissus externes au phloème, et dans les lumens des cellules xylemiennes, contient peu d'unités syringyles (type-S). En conséquence, cette ,lignine de défense' se distingue de la ,lignine de développement' typique d'E. gunnii, qui est composée d'unités guaiacyles (type-G) et d'unités syringyles (type-S). En revanche chez les plantules blessées, des unités syringyles sont déposées dans les parois des cellules immatures du xylème à un stade où les cellules comparables des plantules témoins ne contiennent que très peu d'unités syringyles. La deuxième approche a indiqué que deux activités alcools cinnamyliques déshydrogénases différentes sont induites, et régulées d'une façon différencielle, en réponse à la blessure chez les arbres d'E. gunnii. L'activité alcool coniférylique déshydrogénase est induite rapidement et continue d'augmenter pendant les 15 premiers jours de la période expérimentale de 17 jours, tandis que l'activité alcool sinapylique déshydrogénase est seulement détectée à 8 jours après la blessure et continue d'augmenter le long de la période expérimentale. Les rôles biologiques potentiels de ces deux activités alcools déshydrogénases sont discutés en relation avec la formation de la lignine de défense et avec la lignine de développement chez les arbres. Zusammenfassung Zur Untersuchung der Ligninbildung nach Verletzungen bei Bäumen wurden zwei sich ergänzende Forschungsansätze gewählt: 1. Mikroskopische und histochemische Untersuchungen der Wundreaktion an Jungpflanzen (3,5 Monate alt) von Eucalyptus gunnii und 2. Biochemische Untersuchungen der Hydroxycinnamylalkohol-Dehydrogenase-Aktivität bei verletzten, sechs Jahre alten E. gunnii -Bäumen im Freiland. Der erste Ansatz zeigte, dass eine Barrierezone als Antwort auf die Verletzung sowohl in beiden Grundgeweben (Cortex-Barriere und Reaktionszone im Mark) als auch in den Leitgeweben gebildet wird. Die Barrierezone war 24 Stunden nach der Verletzung gerade erkennbar, nach sieben Tagen war sie gut entwickelt. Die mikroskopische Untersuchung zeigte, dass die Barrierezone durch Verstärkung der Zellwände mit ,,ligninartigem Material,, im Grund- und Leitgewebe gebildet wurde, und dass zusätzlich das Lumen gewisser Xylemzellen (Gefässe und Fasern) durch Ablagerung von polymerem phenolischem Material verschlossen wurde. Die histochemische Analyse ergab, dass das ligninartige Material (,,Abwehrlignin,,), das in den Zellwänden des Grundgewebes und in den Lumina der Xylemzellen abgelagert wurde, geringe Mengen an Syringyl-(S-Typ)-Lignineinheiten enthielt und sich somit von der normalen Guaiacyl-Syringyl(G-S)-Komposition des Lignins von E. gunnii unterschied. Das S-Typ-Lignin wurde offenbar in den Zellwänden sich entwickelnder sekundärer Xylemzellen abgelagert. Diese Einlagerung erfolgte in einem Stadium, in dem die Zellwände der vergleichbaren Zellen in unverletzten Kontrollstämmen Lignin mit geringem Syringylgehalt enthielten. Der zweite Versuchsansatz zeigte, dass als Reaktion auf die Verletzung zwei verschiedene Arten von Cinnamylalkohol-Dehydrogenase-Aktivitäten induziert und offensichtlich unterschiedlich reguliert werden. Die Aktivität der Coniferyl-Alkohol-Dehydrogenase wurde sofort induziert und sie nahm während 15 Tagen der 17tägigen Versuchsperiode stetig zu, während die Aktivität der Sinapyl-Dehydrogenase erstmals 8 Tage nach der Verletzung nachweisbar war und dann während der gesamten Versuchsperiode anstieg. Die biologische Bedeutung der beiden Alkoholdehydrogenase-Aktivitäten werden in Bezug auf die Bildung von Abwehr-Lignin im Vergleich zur normalen Ligninbildung in Bäumen diskutiert. [source] Studies on Cell Number and Nuclei in Spores and on Ploidy Level in Ascochyta rabiei IsolatesJOURNAL OF PHYTOPATHOLOGY, Issue 5 2001Bruns Isolates of the phytopathogenic ascomycete Ascochyta rabiei (Pass.) Labr. were stained with the DNA-specific fluorochrome 4,,6-diamidino-2-phenylindole (DAPI) and compared for differences in number of nuclei per pycnidiospore and the ploidy level. Microscopic analyses revealed that within the examined isolates five different combinations of cell number and number of nuclei in spores exist. A one-celled spore may contain one, two and four nuclei, respectively, and in the case of two-celled spores there exist types with one and two nuclei in one cell. Microfluorometric analyses of wild types and benomyl-treated isolates revealed differences in ploidy level among the wild types. [source] Magnaporthe grisea interactions with the model grass Brachypodium distachyon closely resemble those with rice (Oryza sativa)MOLECULAR PLANT PATHOLOGY, Issue 4 2004ANDREW P. M. ROUTLEDGE SUMMARY Germplasm of Brachypodium distachyon was inoculated with Magnaporthe grisea using either rice- (Guy11) or grass-adapted (FAG1.1.1, PA19w-06, PA31v-01) host-limited forms of the fungus, and interactions with varying degrees of susceptibility and resistance were identified. Ecotype ABR5 was resistant to each M. grisea strain whereas ABR1 was susceptible to all but P31vi-01. Mendelian segregation in ABR1 × ABR5 crosses suggested that a single dominant resistance gene conferred resistance to Guy11. Microscopic analyses revealed that the aetiology of Guy11 fungal development and disease progression in ABR1 closely resembled that of rice infections. In ABR5, Guy11 pathogenesis was first suppressed at 48 h post-inoculation, at the secondary hyphal formation stage and was coincident with cytoplasmic granulation. Resistance to strains PA31v-01 and FAG1.1.1 was associated with a localized cell death with little callose deposition. 3,3-Diaminobenzidine staining indicated the elicitation of cell death in B. distachyon was preceded by oxidative stress in the interacting epidermal cells and the underlying mesophyll cells. Northern blot hybridization using probes for barley genes (PR1, PR5 and PAL) indicated that each was more rapidly expressed in ABR5 challenged with Guy11 although the B. distachyon defence genes BD1 and BD8 were more quickly induced in ABR1. Such data show that B. distachyon is an appropriate host for functional genomic investigations into M. grisea pathology and plant responses. [source] Phenotypic and genetic analysis of the Triticum monococcum,Mycosphaerella graminicola interactionNEW PHYTOLOGIST, Issue 4 2008Hai-Chun Jing Summary ,,Here, the aim was to understand the cellular and genetic basis of the Triticum monococcum,Mycosphaerella graminicola interaction. ,,Testing for 5 yr under UK field conditions revealed that all 24 T. monococcum accessions exposed to a high level of natural inocula were fully resistant to M. graminicola. When the accessions were individually inoculated in the glasshouse using an attached leaf seeding assay and nine previously characterized M. graminicola isolates, fungal sporulation was observed in only three of the 216 interactions examined. Microscopic analyses revealed that M. graminicola infection was arrested at four different stages post-stomatal entry. When the inoculated leaves were detached 30 d post inoculation and incubated at 100% humidity, abundant asexual sporulation occurred within 5 d in a further 61 interactions. ,,An F2 mapping population generated from a cross between T. monococcum accession MDR002 (susceptible) and MDR043 (resistant) was inoculated with the M. graminicola isolate IPO323. Both resistance and in planta fungal growth were found to be controlled by a single genetic locus designated as TmStb1 which was linked to the microsatellite locus Xbarc174 on chromosome 7Am. ,,Exploitation of T. monococcum may provide new sources of resistance to septoria tritici blotch disease. [source] Involvement of hydrogen peroxide in leaf abscission signaling, revealed by analysis with an in vitro abscission system in Capsicum plantsTHE PLANT JOURNAL, Issue 1 2008Masaru Sakamoto Summary Although auxin and ethylene play pivotal roles in leaf abscission, the subsequent signaling molecules are poorly understood. This is mainly because it is difficult to effectively treat the intact abscission zone (AZ) with pharmacological reagents. We developed an in vitro experimental system that reproduces stress-induced leaf abscission in planta. In this system, 1-mm-thick petiole strips, encompassing the AZ, were separated within 4 days of abscission at the AZ through cell wall degradation in an auxin depletion- and ethylene-dependent manner. The system allowed us to show that hydrogen peroxide (H2O2) is involved in abscission signaling. Microscopic analyses revealed continuous H2O2 production by AZ cells. H2O2 scavengers and diphenylene iodonium, an inhibitor of NADPH oxidase, suppressed in vitro abscission and cellulase expression. Conversely, the application of H2O2 promoted in vitro abscission and expression of cellulase. Ethephon-induced abscission was suppressed by inhibitors of H2O2 production, whereas the expression of ethylene-responsive genes was unaffected by both H2O2 and an H2O2 inhibitor. These results indicated that H2O2 acts downstream from ethylene in in vitro abscission signaling. In planta, salinity stress induced the expression of genes that respond to ethylene and reactive oxygen species, and also induced H2O2 production at the AZ, which preceded leaf abscission. These results indicate that H2O2 has roles in leaf abscission associated with ethylene both in vitro and in planta. [source] Ultrasonic treatment of waste activated sludgeENVIRONMENTAL PROGRESS & SUSTAINABLE ENERGY, Issue 2 2006Raf Dewil Abstract Activated sludge processes are key technologies to treat wastewater. These biological processes produce huge amounts of waste activated sludge (WAS), now commonly called biosolids. Mechanical, thermal, and/or chemical WAS conditioning techniques have been proposed to reduce the sludge burden. The ultrasonic treatment of WAS is quite novel. The present paper reports on extensive investigations using an ultrasonic treatment of WAS, to study its potential to meet one or all of four objectives: (1) reduce WAS quantities; (2) achieve a better dewaterability; (3) provoke a release of soluble chemical oxygen demand (COD) from the biosolids, preferably transformed into biodegradable organics; and (4) possibly destroy the filamentous microorganisms responsible for sludge bulking. Although meeting these objectives would help to solve the problems cited, the energy consumption could be a considerable drawback: the paper will thus assess whether all or some objectives are met, and at what operational cost. A literature survey defines the occurring phenomena (cavitation) and the important operation parameters [such as frequency, duration, specific energy input (SE)]. The experiments are carried out in a batch reactor of volume up to 2.3 L. The ultrasonic equipment consisted of a generator, a converter, and a sonotrode, supplied by Alpha Ultrasonics under the brand name of Telsonic. Three different kinds of sludge were tested, with different concentrations of dry solids (DS) between approximately 3.5 and 14 g DS/L WAS. Ultrasonic energy was introduced in a continuous manner (against possible pulsed operation). The major operational parameters studied include duration of the ultrasonic treatment and specific energy input. The applied frequency was set at 20 kHz. The release of COD from the WAS phase into the filtrate phase is a function of the specific energy input with yields of nearly 30% achievable at SE values of 30,000 kJ/kg DS. A major fraction of the COD is transformed into biodegradable organics (BOD). The reduction in DS fraction of the sludge is proportional to the COD release rates. Although the DS content is reduced, the dewaterability of the sludge is not improved. This reflects itself in increased filtration times during vacuum filtration and in increased values of the capillary suction time (CST). This more difficult dewaterability is the result of considerably reduced floc sizes, offering an extended surface area: more surface water is bound (CST increases) and the filterability decreases as a result of clogging of the cake. To reach the same dryness as for the untreated cake, the required dosage of polyelectrolyte is nearly doubled when the SE of the ultrasound treatment is increased from 7500 to 20,000 kJ/kg DS. The ultrasonic reduction of filamentous WAS organisms is not conclusive and very little effect is seen at low intensities and short treatment durations. Microscopic analysis of the WAS identified the dominant presence of Actynomyces. The release of soluble COD and BOD certainly merit further research. © 2006 American Institute of Chemical Engineers Environ Prog, 2006 [source] First report of saxitoxin in Finnish lakes and possible associated effects on human healthENVIRONMENTAL TOXICOLOGY, Issue 3 2005Jarkko Rapala Abstract This study is the first report of saxitoxin in cyanobacterial blooms in Finland. Bloom samples (n = 50) were collected from Finnish freshwater sites during summer months of 2002 and 2003. These samples were screened for the presence of paralytic shellfish toxins (PSTs) using the Jellett rapid PSP screening test. Samples testing positive for PSTs (n = 7) were further analyzed with saxiphilin- and voltage-gated sodium channel [3H]-STX,binding radioreceptor assays and liquid chromatography using fluorescence and mass spectrometric analysis. The results indicated that saxitoxin (STX) was the only PST analogue in the samples and that it was present in high concentrations, as much as 1 mg L,1. Microscopic analysis revealed that 95%,100% of the phytoplankton in the positive samples consisted of Anabaena lemmermannii. The trophic status of lakes in which STX-containing blooms were found varied from oligotrophic to hypertrophic. All the lakes had high nitrogen-to-phosphorus ratios. In some instances, samples had been collected from sites where swimmers had reported adverse health effects, and in three such cases, reported adverse health effects were associated with sites from which samples testing positive for STX had been received. Symptoms of fever, eye irritation, abdominal pains, and skin rash were reported in children aged 2,10 years after exposure to the water. These were not the adverse human symptoms typical of STX poisoning; rather, they represented acute effects often reported following recreational exposure to cyanobacterial blooms. © 2005 Wiley Periodicals, Inc. Environ Toxicol 20: 331,340, 2005 [source] Phenotypic diagnosis of dwarfism in six Friesian horsesEQUINE VETERINARY JOURNAL, Issue 3 2008W. BACK Summary An extreme form of abnormal development, dwarfism, is common in man and some animals, but has not been officially reported in horses. Within the Friesian horse breed, congenital dwarfism has been recognised for many years, but no detailed report exists on its phenotype. The most salient feature of the dwarf syndrome is the physeal growth retardation in both limbs and ribs. Affected animals have approximately 25% shorter fore- and hindlimbs and approximately 50% reduced bodyweight. Post natal growth is still possible in these animals, albeit at a slower rate: the head and back grow faster than the limbs and ribs leading to the characteristic disproportional growth disturbance. Thus, mature dwarfs exhibit a normal, but a relatively larger head conformation, a broader chest with narrowing at the costochondral junction, a disproportionally long back, abnormally short limbs, hyperextension of the fetlocks and narrow long-toed hooves. Furthermore, a dysplastic metaphysis of the distal metacarpus and metatarsus is radiographically evident. Microscopic analysis of the growth plates at the costochondral junction shows an irregular transition from cartilage to bone, and thickening and disturbed formation of chondrocyte columns, which is similar to findings in osteochondrodysplasia. [source] Hen Egg Yolk Prevents Bacterial Adherence: A Novel Function for a Familiar FoodJOURNAL OF FOOD SCIENCE, Issue 1 2001T. Deignan ABSTRACT: The aim of this study was to evaluate the ability of unfractionated hen egg yolk to prevent the attachment of Salmonella typhimurium to murine intestinal epithelial cells in vitro. Inhibition of adherence was examined microscopically and by an ELISA. Both methods showed that egg yolk from immunized and unimmunized hens significantly reduced bacterial adherence. Optical density (OD) readings ± SD of 0.825 ± 0.016 for untreated bacteria were reduced to 0.335 ± 0.024 and 0.267 ± 0.021 for bacteria pretreated with egg yolk from immunized hens and unimmunized hens, respectively. Microscopic analysis showed that egg yolk from either immunized or unimmunized hens reduced bacterial adherence from 17 ± 2.7 bacteria per epithelial cell to less than 4 bacteria per epithelial cell. These results show that hen egg yolk significantly inhibits adherence of S. typhimurium to intestinal epithelial cells in vitro. Boosting of specific antibody levels does not enhance this antiadhesive effect, suggesting that egg yolk contains novel antiadhesive factors. [source] Psoriasis under the microscopeJOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY & VENEREOLOGY, Issue 2006BJ Cribier Abstract Histopathology is a major diagnostic tool in dermatology, particularly in psoriasiform diseases. Morphological studies showed that the initial event in psoriatic lesions is perivascular infiltrate, followed by dilatation of superficial papillary vessels. Proliferation of keratinocytes and neutrophil exocytosis are secondary events. Fully developed psoriasis has a very characteristic pattern, which includes elongation of rete ridges leading to regular acanthosis, oedema of the papillary dermis associated with tortuous dilated vessels, thinning of suprapapillar area, decreased thickness of granular layer, and exocytosis of neutrophils in the spinous layer (Kogoj's pustule) or in the cornified parakeratotic layer (Munro microabscesses). Pustular psoriasis is characterized by large or confluent intra-epidermal multilocular pustules. Whatever the clinical variant of psoriasis, common morphological signs suggest that it is basically a unique pathological process, with many possible presentations according to various factors such as age, size and localization of lesions, or therapy. Similar microscopic elementary lesions indicate that Hallopeau's acrodermatitis continua, Reiter's disease and geographical tongue are variants of psoriasis. Because of the many faces of the disease, psoriasis can resemble many other squamous or pustular disorders. Differential diagnosis by microscopic analysis is based on pattern analysis, PAS (Periodic Acid Schiff) staining to rule out fungal infection, and immunohistochemistry to characterize lymphocytic infiltrate. Psoriasis is one of the most common inflammatory skin diseases. In its characteristic presentation, psoriasis comprises well-circumscribed red scaly papules and plaques. In this form, the disease is generally easy to identify, especially when the elbows, knees and scalp are affected. Nevertheless, the term ,psoriasis' includes more clinical variants than any other inflammatory dermatosis: psoriasis vulgaris vs. pustular, localized vs. generalized, topographic variants, mucous membranes involvement, hair and nail lesions. Although some of these conditions might be extremely different from psoriasis vulgaris, common pathological findings can be identified in all of them. Microscopic analysis of psoriatic lesions may therefore help clinicians to make the diagnosis and to understand that, whatever the clinical presentation, signs and symptoms are mainly due to a unique pathological process. [source] Microglial colonization of the developing mouse brain: the effect of CD11b deletionNEUROPATHOLOGY & APPLIED NEUROBIOLOGY, Issue 2 2002J. K. Jeetle Introduction:, Microglia are resident mononuclear phagocytes of the central nervous system, which colonize the brain both prenatally and after birth. It is proposed that they enter the brain initially via the surrounding mesenchyme, via ventricles and later through blood vessels, but the mechanisms of entry and signals used for migration are still to be established. Previous studies have shown that ligands for some integrin adhesion molecules expressed on blood vessels in the developing nervous system (particularly ICAM-1 and ICAM-2 which bind CD11a/LFA-1 and CD11b/Mac-1), may act as potential recruiting signals for microglial precursors. This study addressed whether CD11b is influential on the migration of microglial precursors into the developing CNS. Material and methods:,Ricinus communis agglutinin-1 (RCA-1) lectin histochemistry was employed to anatomically map the distribution of amoeboid and ramified microglia from embryonic day 15 (E15) to birth. Embryonic mouse brains from CD11b knockout (,/,) (n = 42), and heterozygote (+/,) (n = 52) mice generated on a C57/BL6 background (Melo et al. Cell Immunol 2000; 205: 13,23) and wild-type (+/+) (n = 37) litter mates were fixed in Bouin's solution, processed to paraffin wax and serially sectioned at 15,40 µm. To investigate further potential signals for recruiting microglial precursors, brains were immunochemically screened for integrins CD11a, CD11b, CD18, ,X, VLA-4 and the chemokine MCP-1. Results:, Microscopic analysis revealed the morphological transition of microglia from predominantly amoeboid forms at E15,E16 to a flourishing population of ramified cells at E19,E20. RCA-1 histochemistry showed no clear differences in microglial distribution or timing of colonization between CD11b (,/,) and wild-type mice from E15 to birth. Although CD11b deletion did not influence the timing of microglial ramification, there appeared to be fewer ramified cells in (,/,) mice within comparative brain regions. This requires further quantitative morphometric analysis. Of the integrins investigated, none were restricted to microglia and only VLA-4 and ,X showed reactivity within the CNS. However, MCP-1 was notably localized to the cortical plate within all genotypes, consistent with previous findings in human foetal CNS (Rezaie & Male. Microsc Res Tech 1999; 45: 359,382). Conclusion:, The results suggest that CD11b has little influence on the timing or regional distribution of microglia in the developing murine CNS. It is more likely that CD11b is only one of several factors that influence the migration and differentiation of these cells. [source] Microscopic analysis and seasonality of gemma production in the freshwater red alga Hildenbrandia angolensis (Hildenbrandiales, Rhodophyta)PHYCOLOGICAL RESEARCH, Issue 4 2000Alison R. Sherwood SUMMARY The development and release of the unique vegetative propagules of the freshwater encrusting alga Hildenbrandia angolensis Welwitsch ex West et West, gemmae, were studied using several different microscopic and histochemical techniques. In addition, the seasonality of gemma production was monitored bimonthly over a 12-month period in two spring-fed streams in Texas, USA. Gemmae differentiate within the thallus and are subsequently released from the surface of the crust. Release of the gemmae most likely occurs by digestion of surrounding cells, as suggested by the presence of starch granules and lipid globules in the region between the released gemma and the thallus. The initial separation of the gemmae from the thallus occurs from the sides of the gemma or the bottom, or possibly simultaneously. Contrary to previous studies, we have observed that gemma production occurs endogenously within the thallus of freshwater Hildenbrandia, rather than on the surface of the crust in raised structures. Histochemical tests and electron microscopic examination indicate that the cells of the gemmae contain a large amount of floridean starch. The starch granules frequently form rings surrounding the nuclei of both gemma and thallus cells; a feature infrequently reported for florideophyte red algae. Our seasonality investigations indicate that large fluctuations in gemma production occur over 1 year, but at least some gemma production continues year-round in the streams examined. [source] Percolation phenomena in carbon black,filled polymeric concretePOLYMER ENGINEERING & SCIENCE, Issue 9 2000L. Rejón Percolation in carbon black-filled polymeric concrete, is discussed based on the measured changes in electrical conductivity and morphology of the composite at different concentrations of carbon black. The percolation threshold ranged between 6 and 7 wt% (based on resin weight) of carbon black. Above this concentration, the filler particles formed agglomerates in contact with each other, suggesting that the conduction process is nearly ohmic in nature. A power law predicted by percolation theory described the behavior of the conductivity as a function of carbon black content. Microscopic analysis showed the presence of a continuous structure formed by the polyester resin and carbon black, in which silica particles were embedded. [source] Isolation, Partial Purification, and Immunogenicity of Flagella from Tritrichomonas foetusTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 3 2005PHILIP F. JEMILOHUN Abstract. Tritrichomonas foetus, the agent of bovine trichomoniasis, is a flagellate protozoan responsible for substantial economic losses to the dairy and calf industries worldwide. As yet, there is no approved treatment nor is there a sensitive diagnostic method. All these problems suggest that immunization is the best control strategy. In view of this, we isolated and partially purified flagella of the parasite by vortex homogenization followed by low-speed differential centrifugation. The resulting enriched flagellar preparation termed "crude flagellar prep" was purified further by sucrose and percoll gradients. Microscopic analysis showed that the flagellar membrane was intact. Analysis by sodium dodecyl-sulfate polyacrylamide gel electrophoresis revealed three prominent protein bands of 42, 49, and >250 kDa, and several minor bands. Immunoblotting of flagellar and whole-cell extracts revealed many flagellar antigens. [source] A subtilisin-like serine protease essential for mucilage release from Arabidopsis seed coatsTHE PLANT JOURNAL, Issue 3 2008Carsten Rautengarten Summary During Arabidopsis seed development large quantities of mucilage, composed of pectins, are deposited into the apoplast underneath the outer wall of the seed coat. Upon imbibition of mature seeds, the stored mucilage expands through hydration and breaks the outer cell wall that encapsulates the whole seed. Mutant seeds carrying loss-of-function alleles of AtSBT1.7 that encodes one of 56 Arabidopsis thaliana subtilisin-like serine proteases (subtilases) do not release mucilage upon hydration. Microscopic analysis of the mutant seed coat revealed no visible structural differences compared with wild-type seeds. Weakening of the outer primary wall using cation chelators triggered mucilage release from the seed coats of mutants. However, in contrast to mature wild-type seeds, the mutant's outer cell walls did not rupture at the radial walls of the seed coat epidermal cells, but instead opened at the chalazal end of the seed, and were released in one piece. In atsbt1.7, the total rhamnose and galacturonic acid contents, representing the backbone of mucilage, remained unchanged compared with wild-type seeds. Thus, extrusion and solubility, but not the initial deposition of mucilage, are affected in atsbt1.7 mutants. AtSBT1.7 is localized in the developing seed coat, indicating a role in testa development or maturation. The altered mode of rupture of the outer seed coat wall and mucilage release indicate that AtSBT1.7 triggers the accumulation, and/or activation, of cell wall modifying enzymes necessary either for the loosening of the outer primary cell wall, or to facilitate swelling of the mucilage, as indicated by elevated pectin methylesterase activity in developing atsbt1.7 mutant seeds. [source] Mutants in DEFECTIVE GLYCOSYLATION, an Arabidopsis homolog of an oligosaccharyltransferase complex subunit, show protein underglycosylation and defects in cell differentiation and growthTHE PLANT JOURNAL, Issue 4 2005Olivier Lerouxel Summary A mutant called defective glycosylation1-1 (dgl1-1) was identified in Arabidopsis based on a growth defect of the dark-grown hypocotyl and an abnormal composition of the non-cellulosic cell wall polysaccharides. dgl1-1 is altered in a protein ortholog of human OST48 or yeast WBP1, an essential protein subunit of the oligosaccharyltransferase (OST) complex, which is responsible for the transfer in the ER of the N-linked glycan precursor onto Asn residues of candidate proteins. Consistent with the known function of the OST complex in eukaryotes, the dgl1-1 mutation led to a reduced N-linked glycosylation of the ER-resident protein disulfide isomerase. A second more severe mutant (dgl1-2) was embryo-lethal. Microscopic analysis of dgl1-1 revealed developmental defects including reduced cell elongation and the collapse and differentiation defects of cells in the central cylinder. These defects were accompanied by changes in the non-cellulosic polysaccharide composition, including the accumulation of ectopic callose. Interestingly, in contrast to other dwarf mutants that are altered in early steps of the N -glycan processing, dgl1-1 did not exhibit a cellulose deficiency. Together, these results confirm the role of DGL1 in N-linked glycosylation, cell growth and differentiation in plants. [source] Immunoexpression of Cbfa-1/Runx2 and VEGF in sinus lift procedures using bone substitutes in rabbitsCLINICAL ORAL IMPLANTS RESEARCH, Issue 6 2010Leandro Soeiro De Souza Nunes Abstract Objectives: To analyze and compare the expression of core binding factor-1 (Cbfa-1)/Runx2 and vascular endothelium growth factor (VEGF) in sinus lift procedures using bovine hydroxyapatite (HA) and ,-tricalcium phosphate (,-TCP). Material and Methods: Twenty-four male rabbits that had undergone bilateral sinus lift procedures were divided into three groups, according to the sinus filling material: Group 1: autogenous bone graft; Group 2: bovine HA; and Group 3: ,-TCP. All groups were sacrificed after 7, 14, 30 and 60 days, for microscopic, histomorphometry and immunohistochemistry analysis. Results: Microscopic analysis showed a similar bone repair pattern between the tested groups. New bone formation, soft and medular tissue, remaining material or particulate bone graft area were obtained by histomorphometric analysis. After 14 days, statistically significant differences in new bone formation were found between Group 1 (27.76±7.8) and Groups 2 (14.22±3.2) and 3 (11.1±7.7). After 30 days, statistically significant differences (P<0.05) were detected in bone formation between Groups 1 (31.39±36.5) and 2 (14.13±3.2). The last period showed improved bone formation in Group 2. Also, Group 2 showed higher Cbfa-1/Runx2 immunoexpression when compared with Group 3. No remarkable differences were observed in VEGF immunoexpression among groups. Conclusion: Taken together, both biomaterials allowed bone tissue growth in a conductive pattern and did not interfere with bone remodeling in the late period, with a slight improvement in bone tissue formation when using HA, confirmed by marked expression of Cbfa-1 at initial periods. To cite this article: Nunes LSS, De Oliveira RV, Holgado LA, Nary Filho H, Ribeiro DA, Matsumoto MA. Immunoexpression of Cbfa-1/Runx2 in sinus lift procedures using bone substitutes in rabbits. Clin. Oral Impl. Res. 21, 2010; 584,590. doi: 10.1111/j.1600-0501.2009.01858.x [source] Effect of chronic denervation and denervation-reinnervation on cytoplasmic creatine kinase transcript accumulationDEVELOPMENTAL NEUROBIOLOGY, Issue 3 2001Charles H. Washabaugh Abstract The extensor digitorum longus (EDL) and soleus muscles of adult mice were chronically denervated or denervated and allowed to reinnervate. Muscles were evaluated 1, 5, 14, 21, and 52 days after sciaticectomy. In terms of weight loss, myofiber atrophy, degeneration, and fibrosis, the soleus muscle was more affected than the EDL by chronic denervation. Fifty-two days after chronic denervation, the number of molecules of MCK/ng total RNA in both muscles (determined with competitive PCR) decreased, with the soleus muscle being more affected. At that stage, BCK mRNA levels in the denervated soleus were unchanged, but they were increased (>50%) in the EDL. Reinnervation restored MCK transcript accumulation in the EDL, whereas, in the soleus MCK, transcripts exceeded control values by 57%, approaching levels in the reinnervated EDL. Despite restoration of MCK mRNA levels, the number of molecules of BCK mRNA/ng total RNA was four- to fivefold higher in reinnervated versus control muscles, suggesting that the genes encoding the CK mRNAs are not coordinately regulated in adult muscle. The role of denervation induced, fiber type changes in regulating CK mRNA accumulation has been evaluated. Electron microscopic analyses have established that fibrosis is not a factor that determines BCK mRNA levels in the chronically denervated or denervated-reinnervated muscles. CK isozyme analyses support the hypothesis that a greater proportion of BCK mRNA found in 52 day chronically denervated and denervated-reinnervated muscles is produced in myofibers vs. nonmuscle cells than in control muscles. © 2001 John Wiley & Sons, Inc. J Neurobiol 47: 194,206, 2001 [source] The characterization of functions involved in the establishment and maturation of Klebsiella pneumoniae in vitro biofilm reveals dual roles for surface exopolysaccharidesENVIRONMENTAL MICROBIOLOGY, Issue 3 2008Damien Balestrino Summary The ability to form biofilm is seen as an increasingly important colonization strategy among both pathogenic and environmental Klebsiella pneumoniae strains. The aim of the present study was to identify abiotic surface colonization factors of K. pneumoniae using different models at different phases of biofilm development. A 2200 K. pneumoniae mutant library previously obtained by signature-tagged mutagenesis was screened in static and dynamic culture models to detect clones impaired at early and/or mature stages of biofilm formation. A total of 28 mutants were affected during late phases of biofilm formation, whereas 16 mutants displayed early adhesion defect. These mutants corresponded to genes involved in potential cellular and DNA metabolism pathways and to membrane transport functions. Eight mutants were deficient in capsule or LPS production. Gene disruption and microscopic analyses showed that LPS is involved in initial adhesion on both glass and polyvinyl-chloride and the capsule required for the appropriate initial coverage of substratum and the construction of mature biofilm architecture. These results give new insight into the bacterial factors sequentially associated with the ability to colonize an abiotic surface and reveal the dual roles played by surface exopolysaccharides during K. pneumoniae biofilm formation. [source] Use of a novel nonantibiotic triple marker gene cassette to monitor high survival of Pseudomonas fluorescens SBW25 on winter wheat in the fieldFEMS MICROBIOLOGY ECOLOGY, Issue 2 2008Lotta Jäderlund Abstract Pseudomonas fluorescens SBW25 was tagged with a triple marker gene cassette containing gfp, encoding green fluorescent protein; luxAB, encoding luciferase; and telABkilA, encoding tellurite resistance, and the tagged strain was monitored in the first Swedish field release of a genetically modified microorganism (GMM). The cells were inoculated onto winter wheat seeds and the GMM cells (SBW25,tgl) were monitored in the field from September 2005 to May 2006 using plating, luminometry and microscopic analyses. Cell numbers were high on all sampling occasions and metabolically active cells were detected on all plant parts. Field results were similar to those obtained in a parallel phytotron study, although the amount of SBW25,tgl detected on shoots was significantly higher in the phytotron than in the field. After winter, cell counts were 100-fold higher on the roots and root-associated soil compared with prewinter measurements, although the cells had a lower relative metabolic activity. The wheat seeds were naturally infested with Microdochium nivale, but no treatment resulted in reduction of disease symptoms. No SWB25,tgl cells were ever found in bulk soil or uninoculated plants. The Swedish field trial results complement and contrast with prior field studies performed with the same parent organism in the United Kingdom under different soil, plant and climatic conditions. [source] Organelle-specific expression of subunit ND5 of human complex I (NADH dehydrogenase) alters cation homeostasis in Saccharomyces cerevisiaeFEMS YEAST RESEARCH, Issue 6 2010Wojtek Steffen Abstract The ND5 component of the respiratory complex I is a large, hydrophobic subunit encoded by the mitochondrial genome. Its bacterial homologue, the NDH-1 subunit NuoL, acts as a cation transporter in the absence of other NDH-1 subunits. Mutations in human ND5 are frequently observed in neurodegenerative diseases. Wild type and mutant variants of ND5 fused to GFP or a FLAG peptide were targeted to the endoplasmatic reticulum (ER) or the inner mitochondrial membrane of Saccharomyces cerevisiae, which lacks an endogenous complex I. The localization of ND5 fusion proteins was confirmed by microscopic analyses of S. cerevisiae cells, followed by cellular fractionation and immunostaining. The impact of the expression of ND5 fusion proteins on the growth of S. cerevisiae in the presence and absence of added salts was studied. ER-resident ND5 conferred Li+ sensitivity to S. cerevisiae, which was lost when the E145V variant of ND5 was expressed. All variants of ND5 tested led to increased resistance of S. cerevisiae at high external concentrations of Na+ or K+. The data seem to indicate that ND5 influences the salt homeostasis of S. cerevisiae independent of other complex I subunits, and paves the way for functional studies of mutations found in mitochondrially encoded complex I genes. [source] Syntheses, structures, and supramolecular properties of giant ,-expanded macrocyclic oligothiophenesHETEROATOM CHEMISTRY, Issue 5 2007Masahiko Iyoda Fully conjugated ,-expanded macrocyclic oligothiophenes with 60,-, 90,-, 120,-, 150,-, and 180,-electron systems (1,5) were synthesized using modified McMurry coupling reaction as a key step. Compound 1 was converted into macrocyclic oligo(2,5-thienylene-ethynylene) 6 using bromination-dehydrobromination procedure. X-ray analysis of 1 revealed a unique molecular and packing structure, reflecting a round, planar shape with nanoscale inner cavity. Interestingly, 2 and 3 self-aggregate in the solid state to form nanowires. The structure of fibrous aggregates was established by scanning electron microscopic and atomic force microscopic analyses. Compounds 1,6 exhibit multi-step reversible redox behavior with fairly low first oxidation potentials, reflecting their cyclic conjugation. Doping of 1,3 with iodine gives semiconductors owing to their ,-donor properties and ,,, stacking ability. © 2007 Wiley Periodicals, Inc. 18:460,466, 2007; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/hc.20337 [source] Preparation of Crystalline-Oriented Titania Photoelectrodes on ITO Glasses from a 2-Propanol,2,4-Pentanedione Solvent by Electrophoretic Deposition in a Strong Magnetic FieldJOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 5 2009Mamiko Kawakita Crystal-oriented and crack-free thin TiO2 films with a good interfacial adhesion on indium,tin oxide glass substrates for photoelectrodes of dye-sensitized solar cells were fabricated by the constant voltage electrophoretic deposition (EPD) method in a strong magnetic field of 12 T generated by a superconducting magnet. A binder-free suspension for the EPD was prepared by dispersing TiO2 in a mixture of 2-propanol and 2,4-pentanedione (acetylacetone). The electrical conductivity, sedimentation rate, and the electrophoretic mobility were measured at varying ratios of the mixed solution. The optimized state of the suspension exhibiting the highest surface charge potential and producing deposits with the highest green density was obtained at the 50:50 mixing ratio. The TiO2 films were characterized by X-ray diffraction and scanning electron microscopic analyses. [source] Phase Evolution in Heat-Treated Si3N4 with Additions of Yb2O3JOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 2 2008Gang Feng Guo The heat treatment of silicon nitride (Si3N4) ceramics with additions of 8, 12, and 16 wt% Yb2O3 was carried out at different temperatures and the evolution of grain boundary (GB) phase was investigated systematically by X-ray diffraction (XRD) as well as scanning electron and transmission electron microscopic analyses. XRD results reveal that the extent and the ease of GB crystallization increase with increasing the Yb2O3 content, and that high heat-treatment temperatures in general favor crystallization of the quaternary compounds such as the Yb4Si2O7N2 phase. These results provide an insight into the GB phase evolution in the Yb-system Si3N4 ceramics subjected to a postsintering heat treatment. [source] A classification of the fibrin network structures formed from the hereditary dysfibrinogensJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 8 2006T. SUGO Summary.,Objective: The main objective was to study the relationships of the molecular defects in 38 dysfibrinogens with their fibrin networks. Methods and results: Scanning electron microscopic analyses revealed that all the fibrins formed under the same conditions had networks composed of either normal thickness fibers or thin fibers, accompanied by a variety of alterations in the network structure and characteristics. We classified these fibrin networks into five classes, designated normal, less-ordered, porous A, porous B and lace-like networks. The dysfibrinogens with defects in fibrinopeptide A release or the E:D binding sites formed normal or less-ordered networks, while those with defects in the D:D association formed porous A networks composed of many tapered terminating fibers, despite having fibers of normal width, and containing many pores or spaces. The porous B and lace-like networks were composed of highly branched thin fibers because of defects in the lateral association among protofibrils, and the major difference between them was the porosity of the porous B networks. All the porous B networks were easily damaged by mechanical stress, whereas the lace-like networks retained high resistance to such stress, indicating that the network strength was not dependent on the fiber width, but on the porosity that led to fragility of the network. Conclusion: Impairment of the D:D association is the major disturbing factor that leads to the formation of porous fibrin networks. The porosity may be introduced by severe impairment of the D:D association, as well as the lateral association, as has often been observed by extra glycosylation or defects in Ca2+ binding. [source] Ultrastructural evidence for communication between intramuscular vagal mechanoreceptors and interstitial cells of Cajal in the rat fundusNEUROGASTROENTEROLOGY & MOTILITY, Issue 1 2008T. L. Powley Abstract, To assess whether afferent vagal intramuscular arrays (IMAs), putative gastrointestinal mechanoreceptors, form contacts with interstitial cells of Cajal of the intramuscular type (ICC-IM) and to describe any such contacts, electron microscopic analyses were performed on the external muscle layers of the fundus containing dextran-labelled diaminobenzidin (DAB)-stained IMAs. Special staining and embedding techniques were developed to preserve ultrastructural features. Within the muscle layers, IMA varicosities were observed in nerve bundles traversing major septa without contact with ICC-IM, contacting unlabelled neurites and glial cells. IMA varicosities were encountered in minor septa in contact with ICC-IM which were not necessarily in close contact with muscle cells. In addition, IMA varicosities were observed within muscle bundles in close contact with ICC-IM which were in gap junction contact with muscle cells. IMAs formed varicosities containing predominantly small agranular vesicles, occasionally large granular vesicles and prejunctional thickenings in apposition to ICC-IM processes, indicating communication between ICC and IMA via synapse-like contacts. Taken together, these different morphological features are consistent with a hypothesized mechanoreceptor role for IMA-ICC complexes. Intraganglionic laminar ending varicosities contacted neuronal somata and dendrites in the myenteric plexus of the fundus, but no contacts with ICC associated with Auerbach's plexus were encountered. [source] The Lmgpi15 gene, encoding a component of the glycosylphosphatidylinositol anchor biosynthesis pathway, is required for morphogenesis and pathogenicity in Leptosphaeria maculansNEW PHYTOLOGIST, Issue 4 2008Estelle Remy Summary ,,Random insertional mutagenesis was used to investigate pathogenicity determinants in Leptosphaeria maculans. One tagged nonpathogenic mutant, termed m20, was analysed in detail here. ,,The mutant phenotype was investigated by microscopic analyses of infected plant tissues and in vitro growth assays. Complementation and silencing experiments were used to identify the altered gene. Its function was determined by bioinformatics analyses, cell biology experiments and functional studies. ,,The mutant was blocked at the invasive growth phase after an unaffected initial penetration stage, and displayed a reduced growth rate and an aberrant hyphal morphology in vitro. The T-DNA insertion occurred in the intergenic region between two head-to-tail genes, leading to a complex deregulation of their expression. The unique gene accounting for the mutant phenotype was suggested to be the orthologue of the poorly conserved Saccharomyces cerevisiae gpi15, which encodes for one component of the glycosylphosphatidylinositol (GPI) anchor biosynthesis pathway. Consistent with this predicted function, a functional translational fusion with the green fluorescent protein (GFP) was targeted to the endoplasmic reticulum. Moreover, the mutant exhibited an altered cell wall and addition of glucosamine relieved growth defects. ,,It is concluded that the GPI anchor biosynthetic pathway is required for morphogenesis, cell wall integrity and pathogenicity in Leptosphaeria maculans. [source] Morphological characterization of the interaction between Diplocarpon rosae and various rose speciesPLANT PATHOLOGY, Issue 1 2005O. Blechert Blackspot, caused by Diplocarpon rosae, is the most severe and ubiquitous disease of garden roses, but information is lacking about genotype-specific forms of resistance and susceptibility of the host. Macro- and microscopic analyses of 34 rose genotypes with a defined monoconidial culture black spot inoculum identified susceptible and resistant rose genotypes and further genotype-specific subdivisions, indicating the presence of partial forms of resistance and different resistance mechanisms. In total, eight interaction types were characterized, five representing compatible (types 1,5) and three representing incompatible interactions (types 6,8). The incompatible interactions were characterized by the lack of any visible fungal structures beneath the cuticle (type 8), single-cell necroses (type 7) or necroses of larger cell clusters (type 6), the latter two types with penetration hyphae and haustoria in epidermal cells. [source] Light and Electron Microscopic Analyses of Autologous Pericardial Tissue Used as a Small-Diameter Arterial Graft in DogsARTIFICIAL ORGANS, Issue 1 2002Won Gon Kim Abstract: As a form of small-diameter arterial graft, we implanted fresh autologous pericardium and pericardium treated with 0.6% glutaraldehyde in the bilateral carotid arteries of dogs and then compared the time-related changes of the grafts explanted after the predetermined periods. The pericardial grafts were implanted in 1 animal each for scheduled periods of 3 days, 2 weeks, 1 month, 3 months, and 6 months. The retrieved grafts were processed for light and electron microscopic analyses following gross observation. The glutaraldehyde-treated small-diameter pericardial vascular grafts showed a better endothelialization of the blood-contacting surface and a slower fragmentation of the collagen layers than the fresh grafts although it has yet to be proven whether these differences are so significant as to affect the patency results between the groups. [source] | |||||||||||||||||||||||||||||||||||||||||||||||||