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  • Electron microscopy  (252)
  • Springer  (252)
  • American Chemical Society
  • 2005-2009
  • 1985-1989  (92)
  • 1980-1984  (160)
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Year
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Colloid & polymer science 261 (1983), S. 373-374 
    ISSN: 1435-1536
    Keywords: Electron microscopy ; short-time staining ; nodular structure ; crystallization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
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  • 2
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    Colloid & polymer science 263 (1985), S. 116-119 
    ISSN: 1435-1536
    Keywords: Electron microscopy ; staining ; morphology ; nylon-12 ; orientation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: Abstract The morphology of drawn and annealed sheets of nylon-12 was investigated by transmission electron microscopy of stained sections, and the results compared with equivalent small-angle X-ray scattering (SAXS) patterns. A three-component structure was observed, consisting of crystalline (C) and amorphous (A) regions in the microfibrils and an interfibrillar component whose density was deduced to be intermediate between that of the C and A regions. The crystallite width was given satisfactorily by a Guinier analysis of the SAXS profile.
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  • 3
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    Colloid & polymer science 260 (1982), S. 564-569 
    ISSN: 1435-1536
    Keywords: lin. Polyethylene ; Single crystals ; Heat of Fusion ; DSC ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: Abstract Recently published results for solution crystallized PE single crystals have shown, that the experimental heat of fusionΔH * is higher, if the solvent is exchanged to silicon oil (oil suspension samples) as compared with dried mats. This has been interpreted by the collapse of the original hollow pyramids during drying, inducing lateral defects within the lamellae. The present investigation does not confirm this unexpected result.ΔH * of dried mats (T c 66 to 91 °C) and of the corresponding oil suspension samples agree within the rather small limits of experimental error. The crystallinities as derived fromΔH *, density or WAXS are in excellent agreement. SEM micrographs of cold fractured dried mats show their spongy macromorphology, but TEM micrographs of stained ultra-thin sections reveal the lamellar morphology of the walls, consisting of curved lamellae and stacked hollow pyramides. If a dried mat is sintered at room temperature, a dense transparent film is obtained with a rather regular stacked morphology of large flat lamellae.ΔH * of these films agrees with that of the original mat.
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  • 4
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    Current genetics 14 (1988), S. 163-170 
    ISSN: 1432-0983
    Keywords: Plant mtDNA ; Electron microscopy ; Restriction enzymes ; Hairpin structures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Broad bean (Vicia faba) mitochondrial DNA (mtDNA) includes three circular plasmids: mt-plasmid 1 (1,704 ntp), mt-plasmid 2 (1,695 ntp) and mt-plasmid 3 (1,476 ntp). Partially replicated circular forms of these mt-plasmids have been observed in electron microscope preparations. Restriction enzymes that cleave either mt-plasmid 2 (but not mt-plasmids 1 and 3) or mt-plasmid 3 (but not mt-plasmids 1 and 2) were used to generate linear forms of partially replicated mt-plasmid 2 and mt-plasmid 3 molecules. Analyses of these linearized replicative intermediates, observed by electron microscopy, indicated that in both mt-plasmid 2 and mt-plasmid 3 replication originates at a specific location and proceeds in the same, single direction around the molecules. The replication origins of mt-plasmid 2 and mt-plasmid 3 map close to sequences that can fold into hairpin structures.
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  • 5
    ISSN: 1432-0983
    Keywords: Sugar beet ; Cytoplasmic male sterility ; Mitochondrial DNA ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitochondrial (mt) DNAs from normal (N) and male sterile (S) cytoplasms of sugar been have been isolated and investigated by electron microscopy. The results showed that mtDNA was composed of a heterogeneous population of circular molecules. Their contour lengths varied from 0.28 to 51 μm, but unlike in the case of maize, a large difference was not observed in the distribution of molecular classes greater than 1.0 μm between N and S cytoplasms of sugar beet. On the other hand, N and S cytoplasms were shown to contain their own characteristic combinations of small circular mtDNA species with lengths between 0.28 μm and 0.6 μm. Mitochondrial DNAs from various sources of male-sterile cytoplasms were analyzed by agarose gel electrophoresis to determine the extent of cytoplasmic variation. Additional low molecular weight DNA bands appeared in all male-sterile lines examined, and as a result, three distinctive banding patterns were recognized. These data are in general agreement with those based upon restriction endonuclease digestion of mt and chloroplast DNAs and the genetic analysis of fertility restoration in test crosses.
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  • 6
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    Journal of molecular evolution 26 (1987), S. 347-357 
    ISSN: 1432-1432
    Keywords: Ribosome structure ; Electron microscopy ; Image analysis ; Evolutionary lineages
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Multivariate statistical analysis and classification techniques are powerful tools in sorting noisy electron micrographs of single particles according to their principal features, enabling one to form average images with an enhanced signal-to-noise ratio and a better reproducible resolution. We apply this methodology here to determining the characteristic views of the large (50S) ribosomal subunits from the eubacteriumEscherichia coli and the archaebacteriaMethanococcus vannielii, Sulfolobus solfataricus, andHalobacterium marismortui. Average images were obtained of the subunit in the common crown and kidney projections, but views of the particle in orientations intermediate between these two extremes were also elucidated for all species. These averages show reproducible detail of up to 2.0 nm resolution, thus enabling the visualization and interspecies comparison of many structural features as a first step toward comparing the actual three-dimensional structures. Our results disprove evolutionary lineages recently postulated on the basis of electron microscopical images of ribosomal subunits.
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  • 7
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    Development genes and evolution 191 (1982), S. 205-207 
    ISSN: 1432-041X
    Keywords: Chitin inhibition ; Nikkomycin ; Cuticle ; Electron microscopy ; Epilachna varivestis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleoside antibiotic nikkomycin has proved to be an effective inhibitor of chitin synthesis in the Mexican bean beetleEpilachna varivestis. Ultrastructural investigations show defects in the procuticular area after nikkomycin application which suggest the complete absence of chitin. A cuticle like this is inflexible and too brittle to satisfy its normal function as an exoskeleton. The individuals are not able to free themselves from the exuvia and finally die. Therefore nikkomycin seems to be a potential insecticide with high specifity.
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  • 8
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    Calcified tissue international 33 (1981), S. 143-151 
    ISSN: 1432-0827
    Keywords: Calcium transport ; Cytochalasin B ; Dihydrocytochalasin B ; Colchicine ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In vivo calcium absorption was studied in normal and rachitic chicks. Cytochalasin B (CB) at a concentration of 25 µg/ml added to the medium inside the duodenal lumen inhibited calcium absorption (20 min) from 82.5±1.9% of calcium absorbed in the controls to 59.2±3% in normal and from 70.0±2.3% to 47.0±2.1% in rachitic chicks. In vitro studies by everted ileal sacs of young rabbits also showed an inhibition of active transport of calcium due to CB. Whereas in the controls the ratio of45Ca concentrations in serosal and mucosal media (60 min) was 7.2±0.32, the ratios were 5.24±0.52; 4.40±0.36; 3.40±0.42; 5.77±0.52; 1.38±0.08; and 1.06±0.02 in the presence of CB at concentrations of 5, 10 and 25 µg/ml; colchicine 10−4M, Na citrate 0.02M, and heat-devitalized conditions, respectively.45Ca concentration in the mucosal scrapings was also affected. It showed an increase from controls (15,101±404 cpm/mg) and correlated with CB concentration: 17,378±489, 19,015±1000, and 20,201±362 at 5, 10, and 25 µg/ml, respectively. Dihydrocytochalasin B also inhibited active calcium transport and caused an increase in45Ca concentration in the mucosal scrapings. Correlated electron microscopic studies showed certain changes in the brush border, especially in some actin microfilaments in the terminal web region. It seems that these morphological alterations may be related to transcytoplasmic movement of calcium.
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  • 9
    ISSN: 1432-0827
    Keywords: Alcohol ; Electron microscopy ; Growth plate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary We have previously demonstrated that ethanol has a direct toxic effect on the rat skeleton characterized by decreased trabecular bone volume. In the present study, we examined the ultrastructure of the distal radial epiphyseal growth plates in these same animals. Eight weeks of ethanol administration to 12 male rats results in serum alcohol levels of 140 mg/dl but did not alter the width or light microscopic appearance of the radial growth plate. Quantitative electron microscopy failed to demonstrate morphologic evidence of toxicity in the skeletal cells. We conclude that although ethanol appears to have a direct effect on rat bone characterized by enhanced resorption, toxicity is not attended by ultrastructural changes in the skeletal cells.
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  • 10
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    Calcified tissue international 33 (1981), S. 529-540 
    ISSN: 1432-0827
    Keywords: Bone cells ; Electron microscopy ; PTH ; PGE1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Four different cell populations—designated PF, OB, OC, and PC—were isolated from calvaria of 18-day-old chick embryos for analysis of the effects of hormones on bone tissue. The cell populations were studied with histological and biochemical methods. Apart from the well-known cell types present in calvaria, a new cell type was found in the noncalcified organic matrix between the osteoblastic layer and the calcified matrix. These cells were provisionally called osteocytic osteoblasts. They represent the “transition state” between osteoblasts and osteocytes. On the basis of histological studies with light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM), the PF population was considered to originate primarily from the periosteal fibroblasts, the OB population from the osteoblasts and osteocytic osteoblasts. The population of cells still present in calvaria after removal of periosteal fibroblasts and osteoblasts was called the OC population. This cell population was very much enriched with osteocytes. The fourth isolated population (PC) was a mixed population of fibroblasts, osteoblasts, and preosteoblasts. On exposure to parathyroid hormone (PTH), all four cell populations showed increased lactate production, but only the OB and OC populations displayed increased cAMP production. Prostaglandin E1 (PGE1) stimulated cAMP production in both OB and PF cells. From the results of this study it was concluded that PTH receptors are present on all of the cell types studied, but that occupancy of the receptor induces adenylate cyclase stimulation only in osteocytes and fully differentiated osteoblasts.
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  • 11
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    Calcified tissue international 30 (1980), S. 43-50 
    ISSN: 1432-0827
    Keywords: Osteogenesis ; In vitro ; Electron microscopy ; Mineralization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Chick limb mesenchymal cells differentiate into muscle, cartilage, fibrous, and bone tissue. Previous reports show that when stage 24 limb mesenchymal cells are cultured in vitro, chondrocytes, myocytes, fibrocytes, and osteoblasts can be identified on the basis of morphological and biochemical parameters. The study reported here demonstrates that phenotypic expression in culture seems to be dependent on the initial plating density, Scanning electron microscopic observations indicate that when stage 24 limb mesenchymal cells are initially seeded at high densities (5 × 106 cells per 35 mm culture dish), mounds of cells appear in culture. These mounds represent cartilage nodules composed of a fine fibrous matrix and chondrocytes, surrounded by a loose fibrous connective tissue matrix. Cultures initially plated at intermediate densities (2.0–2.5 × 106 cells/35 mm culture dish) produce a flattened layer of fibrocytes overlying a matrix of collagen fibers and calcium phosphate deposits as determined by electron-microprobe analysis; these observations are indicative of osteoblast expression. Cells seeded at this intermediate density appear larger and possess greater surface area than cells seeded at high density. It is suggested that conditions that permit such increased cell surface area coupled with a relative compaction due to cell crowding may provide conditions permissive for osteogenesis. Based on morphological criteria, it appears that chick limb mesenchymal cell osteogenesis in vitro is not associated with chondrogenesis but represents a separate route of phenotypic expression.
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  • 12
    ISSN: 1432-0827
    Keywords: Rat ; Calvarium ; Electron microscopy ; Preosteoclasts ; Osteoclasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary This is a study of the fine structure of cells of the 20-day fetal rat calvarium. Special attention is given to identifying and characterizing preosteoclasts. These cells are relatively common and located largely, but not exclusively, at the endocranial bone surface. The preosteoclasts are characterized by abundant mitochondria, an incomplete perinuclear Golgi apparatus, and variable-shaped dense granules. The dense granules are unique in appearance in that they contain an internal dense matrix surrounded by a clear halo. Most granules are circular in shape but some are elongate or tubular in form. Granules with identical appearance are observed in osteoclasts. The preosteoclasts are mononucleate, or occasionally binucleate. It is suggested that because preosteoclasts are morphologically distinctive and relatively abundant, it should be feasible to separate these cells from a heterogeneous cell isolate.
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  • 13
    ISSN: 1432-0827
    Keywords: Decalcification ; Electron microscopy ; Bone matrix ; Bone glycoproteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary A solution of triethylammonium EDTA in 80% ethanol was evaluated as a demineralizing reagent for bone in comparison with aqueous solutions of EDTA. Biochemical analysis and acrylamide gel electrophoresis of extracts of finely powdered bovine bone showed that most of the macromolecular components of the organic matrix extractable in aqueous EDTA were retained when the triethylammonium EDTA reagent was used. Ultrastructural examination of chick tibias decalcified with the reagents showed a better preservation of cellular morphology, especially the membranous components, and more uniformly distributed ground substance, though slightly less in quantity, when the aqueous reagent was used. Use of the two reagents appears to be complementary, the alkylammonium reagent being more appropriate for use in studies of the organic matrix of bone, including immunohistochemical studies of bone glycoproteins. The aqueous reagent is more appropriate for use in studies of cellular ultrastructure.
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  • 14
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    Calcified tissue international 33 (1981), S. 119-124 
    ISSN: 1432-0827
    Keywords: Avian eggshell ; Microstructure ; Electron microscopy ; Electron diffraction ; Calcite growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The eggshell of the domestic fowl has been studied by transmission electron microscopy and diffraction. Thin sections of shell were prepared by chemical and ion-beam thinning techniques. Each calcite column of the palisade layer consisted of crystallites of diameter 20 to 30 µm with some tendency for crystallite alignment within a single column. Evidence indicates that there was no significant preferred orientation in the palisade layer as a whole. Only in the surface layer was any preferred orientation detected, and here {1014} planes tended to lie parallel to the surface. The results are compared with previously published data, and calcite nucleation and growth are discussed.
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  • 15
    ISSN: 1432-0983
    Keywords: Electron microscopy ; Replicative intermediates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Some physicochemical properties of the mitochondrial DNAs (mtDNA) from plants of flax, broad bean and mung bean, and from tissue culture cells of jimson weed, soybean, petunia and tobacco were determined. Circular molecules were observed in electron microscope preparations of each mtDNA. In soybean, petunia, broad bean and mung bean mtDNAs, the circular molecules had a continuous distribution of lengths (ranges between 1 to 36 kb, and 1 to 126 kb), heavily skewed toward smaller molecules. Eighty-six percent of the flax circular molecules were from 27 to 54 kb in size, and 78% of the jimson weed circular molecules were from 4 to 15 kb. Replicative forms of 1.2–1.6 kb circular molecules were observed in electron microscope preparations of broad bean mtDNA.
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  • 16
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    European biophysics journal 7 (1981), S. 209-212 
    ISSN: 1432-1017
    Keywords: Photosynthetic bacteria ; Electron microscopy ; Planar lattices
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract The thylakoids of Rhodopseudomonas viridis have been studied by freeze-fracturing whole cells. Depending on growth conditions and treatment before freezing, three different types of particle arrangements in the photosynthetic membrane are reported: a random arrangement, an isometric (quadratic) lattice arrangement with a lattice constant of 12.5 ± 0.8 nm, and a hexagonal lattice arrangement with a lattice constant of 12.5 ± 0.8 nm.
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  • 17
    ISSN: 1432-0983
    Keywords: Allomyces macrogynus ; Mitochondrial DNA ; Electron microscopy ; Restriction enzyme map
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mitochondrial (mt) DNA of the aquatic phycomycete Allomyces macrogynus is a circular molecule with a size of 56.1 kbp. The cleavage sites for the restriction enzymes SalI and PvuI were mapped by comparing the partial denaturation patterns of isolated restriction fragments with the pattern of the intact circle. The genes coding for the small and large ribosomal RNA (rRNA) were located on the restriction map by heteroduplex and R-loop analysis. The gene coding for the large rRNA contains an intervening sequence, app. 0.7 kbp in size, near the 3′-end of the gene. The two rRNA genes are encoded on the same strand of the mtDNA and separated by a region of 17–18 kbp. This rRNA gene organization is similar to that found with members of the Ascomycetes.
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  • 18
    ISSN: 1432-072X
    Keywords: Allomyces ; Zoospores ; Cell wall ; Chitin ; Gamma particle ; Encystment ; Electron microscopy ; Calcofluor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Structural changes during cell wall formation by populations of semisynchronously germinating zoospores were studied in the water mold Allomyces macrogynus. Fluorescence microscopy using Calcofluor white ST (which binds to β-1,4-linked glycans) demonstrated that Calcofluor-specific material was deposited around most cells between 2–10 min after the induction of encystment (beginning when a wall-less zoospore retracts its flagellum and rounds up). During the first 15 min of encystment there was a progressive increase in fluorescence intensity. Ultrastructural analysis of encysting cells showed that within 2–10 min after the induction of encystment small vesicles 35–70 nm diameter were present near the spore surface, and some were in the process of fusing with the plasma membrane. The fusion of vesicles with the zoospore membrane was concomitant with the appearance of electron-opaque fibrillar material outside the plasma membrane. Vesicles similar to those near the spore surface were found within the gamma (γ) particles of encysting cells. These particles had a crystalline inclusion within the electron-opaque matrix. During the period of initial cyst cell wall formation numerous vesicles appeared to arise at the crystal-matrix interface. Approximately 15–20 min was required for the cell wall to be formed. We suggest that the initial response of the zoospore to induction of encystment is the formation of a cell wall mediated by the fusion of cytoplasmic vesicles with the plasma membrane.
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  • 19
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    Archives of microbiology 130 (1981), S. 125-128 
    ISSN: 1432-072X
    Keywords: Rhodospirillum rubrum ; Chromatophores ; Reaction centers ; Liposomes ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In freeze-fractures of chromatophores of Rhodospirillum rubrum the reaction centers are seen as hexagonal arranged particles of 13 nm diameter with a density of around 5,500 particles per μm2. Similar regions on the cytoplasmic membrane suggest that these parts are the prospective invagination sites. Isolated reaction centers are easily incorporated into liposomes. In freeze fractures of liposomes particles similar in shape and size, although less dense as in chromatophores are observed. In negative staining much smaller units of only 5 nm in diameter are found indicating that reaction centers occur in the membrane as tri- or tetramers. There is a strong correlation between particle density in chromatophores and titratable reaction centers remaining in these membranes after extraction of reaction centers by detergents; both values are in good agreement with the yield of reaction centers at a given detergent concentration.
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  • 20
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    Archives of microbiology 142 (1985), S. 333-339 
    ISSN: 1432-072X
    Keywords: Photosynthesis ; Membrane structure ; Electron microscopy ; Photosynthetic bacteria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The organization of photosynthetic membranes in the cytoplasm of the photosynthetic bacterium Rh. viridis has been examined by several techniques for electron microscopy. Thin sections of membrane stacks show that the regular lattice of membrane subunits reported in other studies can be observed in thin section. Tilting of sections in the electron microscope shows that the regular lattices of several membranes overlap in a way that suggests they are in register with each other. This observation can be confirmed by freeze-fracture images in which a regular arrangement of membrane lattices can be observed, each perfectly aligned. Analysis of the spacings of membrane pairs shows that the photosynthetic membranes of Rh. viridis are very closely apposed. The mean diameter of two membranes is 160A, and the average space between two such membranes is only 42A. When a recently developed atomic level model of Rh. viridis reaction center is superimposed against these spacings, each reaction center extends from the surface of its respective membrane far enough to make contact with an apposing membrane. The limited free space between membranes and regular alignment of lattices has a number of implications for how this membrane is organized to carry out the process of energy transfer.
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  • 21
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    Archives of microbiology 146 (1986), S. 267-274 
    ISSN: 1432-072X
    Keywords: Photosynthesis ; Membrane structure ; Electron microscopy ; Ectothiorhodospira ; Serial thin sectioning ; Three dimensional reconstruction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The three dimensional organization of the complete photosynthetic apparatus of the extremely halophilic, bacteriochlorophyll b containing Ectothiorhodospira halochloris has been elaborated by several techniques of electron microscopy. Essentially all thylakoidal sacs are disc shaped and connected to the cytoplasmic membrane by small membraneous “bridges”. In sum, the lumina of all thylakoids (intrathylakoidal space) form one common periplasmic space. Thin sections confirm a paracrystalline arrangement of the photosynthetic complexes in situ. The ontogenic development of the photosynthetic apparatus is discussed based on a structural model derived from serial thin sections.
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  • 22
    ISSN: 1432-072X
    Keywords: Acidaminococcus fermentans ; Glutamate fermentation ; Electron microscopy ; Immunocytochemistry ; Post-embedding labelling ; Antibody-gold complexes ; Protein A-gold complexes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have investigated the in situ location of glutaconyl-CoA decarboxylase and 2-htdroxyglutaryl-CoA dehydratase in Acidaminococcus fermentans using the antibody-gold and protein A-gold techniques carried out as a post-embedding immunoelectron microscopic procedure. Polyclonal antisera were raised in rabbits against homogeneous fractions of the enzymes. Anaerobically grown cells of A. fermentans of the late exponential growth phase were fixed with 0.2% glutaraldehyde and 0.3% formaldehyde (final concentrations) in the growth medium. Dehydration of the cells was achieved with methanol. The cells were embedded in the low temperature embedding resin Lowicryl K4M. The markers indicative for antigenic sites of the two enzymes unequivocally demonstrate that the sodium pump glutaconyl-CoA decarboxylase is located at the cell periphery being a membrane-bound enzyme as expected whereas 2-hydroxyglutaryl-CoA dehydratase is a soluble cytoplasmic enzyme.
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  • 23
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    Archives of microbiology 126 (1980), S. 277-283 
    ISSN: 1432-072X
    Keywords: Bacteriophage ; Myxococcus ; λ ; Superooiled DNA ; Cross-linking ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract DNA was partially released from the heads of myxococcus phages and also coliphage λ and examined by electron microscopy by a modification of the Kleinschmidt technique, in which water was used as hypophase. DNA emerged from the heads in patterns suggestive of newly relaxed supercoils. The unreleased DNA appeared to occupy discrete regions in the head. Some closed circles were released from λ heads. When NaCl solution was used as hypophase, the DNA was observed either released from the tail or from the head, in the latter case, supercoiled regions were observed. When NH4OAc solution was used as hypophase, tightly wound structures were released from λ heads; these fields also contained supercoiled circles. The presence of constrained supercoiled domains in newly released phage DNA was confirmed by observing the effects of ethidium bromide on its conformation. Treatment of phage with nitrogen mustard, a bifunctional alkylating agent, preserved supercoiled domains, even when the phage were lysed over water as hypophase. Further experiments suggested that phage inactivation by nitrogen mustard is largely due to restraint of the supercoiled, native, tertiary structure and that DNA-protein cross-linking may be involved in this reaction. The implications of these findings for the conformation of phage DNA in vivo are discussed and a new model for the winding of DNA in phage heads is proposed.
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  • 24
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    Archives of microbiology 129 (1981), S. 129-134 
    ISSN: 1432-072X
    Keywords: Cockroach ; Hindgut ; Distribution ; Microbial morphotypes ; Transmission ; Electron microscopy ; Statistical analysis ; Eublaberus posticus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The relative numbers of fourteen microbial morphotypes in transmission electron micrographs of the hindgut of a cockroach, Eublaberus posticus, were counted and their distribution was analyzed statistically. The microbiota of three wall-associated regions (the anterior paunch, the posterior paunch, and the black band region) was clearly different from that of the gut lumen. The three wall fractions were also significantly different from each other. Only one of the fourteen types, prosthecate bacteria, appeared to be distributed randomly in the four fractions. The five main wall-associated morphotypes individually constituted up to 41% of the microbes in some micrographs. They included one type with the characteristic morphology of Methanospirillum. Six morphotypes rarely made up over 2% of the population, but were consistently present. The numbers of the remaining three morphotypes were quite variable between micrographs and between individual insects, but when present often made up 5–10% of the population.
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  • 25
    ISSN: 1432-072X
    Keywords: Biological control ; Cyanobacteria ; Electron microscopy ; Entrapment ; lysis ; Myxococcus ; Phormidium ; Spherule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A Myxococcus xanthus isolate from a farm drainage ditch, designated strain PCO2, is capable of rapidly inducing lysis of both agar and liquid-grown cultures of the cyanobacterium, Phormidium luridum, var. olivacea. Microscopic studies of the predator-prey interaction demonstrate that lysis of the cyanobacterium occurs within clumps and spherules formed by the cells of M. xanthus PCO2. In the earliest stage, one sees the formation of irregular microclumps of bacteria and cyanobacterial filaments. As these clumps mature, colonies 1 to 6 mm in diameter develops. The center of these densely green colonies contains cyanohacteria in various stages of degradation, while the periphery is almost exclusively a tightly woven mass of myxobacterial cells. Electron microscopy shows that long extrusions from the outer membrane of the M. xanthus PCO2 cells are involved in the formation both of initial clumps and of mature colonial spherules. These extrusions appear to efficiently entangle the cyanobacterial filaments in the culture environment. Predator-to-prey ratios of 1/10, 1/100 and 1/1,000 have resulted in cyanobacterial lysis. Because the entrapment and lysis of P. luridum filaments by M. xanthus PCO2 appears to be independent of any other heterotrophic nutritional requirement, as well as of environmental agitation, this system has potential as a biological control technique for undesirable aquatic cyanobacteria.
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  • 26
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    Archives of microbiology 138 (1984), S. 273-277 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Secondary metabolite ; Allelopathy ; Photosynthesis ; Electron transport ; Thylakoids ; Herbicides ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cyanobacterin is a secondary metabolite produced by the cyanobacterium, Scytonema hofmanni. Highly purified cyanobacterin was found to inhibit the growth of many cyanobacteria at a minimum effective dose of 2 μg/ml (4.6 μM). The antibiotic had no effect on eubacteria including the photosynthetic Rhodospirillum rubrum. The site of action of cyanobacterin was further investigated in the unicellular cyanobacterium, Synechococcus sp. Electron micrographs of antibiotic-treated Synechococcus cells indicated that cyanobacterin affects thylakoid membrane structure. The antibiotic also inhibited light-dependent oxygen evolution in Synechococcus cells and in spheroplasts. These data support our conclusion that cyanobacterin specifically inhibits photosynthetic electron transport. This activity is similar to herbicides such as 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU). The anhydro analog of cyanobacterin had no biological activity.
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  • 27
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    Archives of microbiology 130 (1981), S. 339-343 
    ISSN: 1432-072X
    Keywords: Leptospira ; Halophilic ; Electron microscopy ; X-ray analysis ; Inclusions ; Cytoplasmic tubules
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    Topics: Biology
    Notes: Abstract The morphology of cells of strain Muggia, a slightly halophilic leptospire, was examined by the negative staining technique. The ultrastructure of the cells was rather similar to that of cells of Leptonema illini, i. e. the cells possessed cytoplasmic tubules. The basal complex of their flagella, however, was similar to the corresponding part of flagella on Gramnegative bacteria. The interior of the cells was densely packed with inclusions, except for the two outermost wavelengths at each end where these inclusions were absent. X-ray microanalysis showed that the inclusions contained sodium and chlorine as their main constituents. The inclusions disappeared upon storage of the cultures at room temperature.
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  • 28
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    Archives of microbiology 132 (1982), S. 10-13 
    ISSN: 1432-072X
    Keywords: Calcofluor White ; Cell wall structure ; Chlorella ; Electron microscopy ; Protoplast ; Ruthenium Red
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Among 12 strains ofChlorella ellipsoidea, C. vulgaris, andC. saccharophila tested, 4 strains (1,C. ellpsoidea; 2,C. vulgaris; 1,C. saccharophila) formed osmotically labile protoplasts after treatment with mixtures of polysaccharide degrading enzymes. The relationship between enzymatical digestibility and structure or composition ofChlorella cell walls were studied by electron microscopy and staining techniques with some specific dyes. The cell wall structures of the 12Chlorella strains were grouped into three types: (1) with a trilaminar outer layer, (2) with a thin outer monolayer, and (3) without an outer layer. Protoplasts were formed only from the strains with a cell wall of Type 2. In the strains with a cell wall of Type 1, the outer layer protected the inner major microfibrillar layer against enzymatic digestion. The cell wall of Type 3 was totally resistant to the enzymes; the chemical composition of the cell wall would be somewhat different from that of other types.
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  • 29
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    Archives of microbiology 135 (1983), S. 169-175 
    ISSN: 1432-072X
    Keywords: Photosynthetic membranes ; Electron microscopy ; Image processing ; Ectothiorhodospira halochloris ; Ectothiorhodospira abdelmalekii ; Rhodopseudomonas viridis ; Rhodopseudomonas sulfoviridis ; Thiocapsa pfennigii
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    Notes: Abstract The photosynthetic membranes of the five bchl b-containing bacteria Ectothiorhodospira halochloris, E. abdelmalekii, Rhodopseudomonas viridis, R. sulfoviridis and Thiocapsa pfennigii have been investigated by electron microscopy and digital image analysis. All five species have the photosynthetic complexes hexagonally arrayed in the membrane with lattice spacings close to 13 nm, except for R. sulfoviridis and T. pfennigii which display somewhat smaller (∼12.5 nm) lattice spacings. Correlation averaging which imposes less stringent requirements on the lattice perfection than conventional Fourier filtration techniques has been employed to elucidate the structure of the photosynthetic complexes. Their basic organization, i.e. a ring, probably containing the light-harvesting (LH) polypeptides, surrounding a core (the “reaction centre”) appears to be almost identical for all species under scrutiny. Despite a resolution of ∼1.6 nm, however, little further significant substructure can be deduced from the averages; possible reasons for the “blurred” appearance of the LH-ring and absence of any subdivision in the reaction centre are discussed along with strategies aimed at obtaining a more detailed model of the molecular architecture of the photosynthetic membranes.
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  • 30
    ISSN: 1432-072X
    Keywords: Bacteriolysis ; Penicillin ; Autolysis ; Cell wall ; Electron microscopy ; Staphylococcus aureus
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    Topics: Biology
    Notes: Abstract The actual reason for the penicillin-induced bacteriolysis of staphylococci was shown to be the “punching” of one or a few minute holes into the peripheral cell wall at predictable sites. These perforations were the result of the lytic activity of novel, extraplasmatic vesicular structures, located exclusively within the bacterial wall material, which we have named “murosomes”. In untreated staphylococci the punching of holes into the peripheral wall is a normal process which follows cross wall completion and represents the first visible step of cell separation. Under penicillin, however, analogous holes are punched by the murosomes at sites of presumptive cell separation even if no sufficient cross wall material had been assembled before at this site (but had rather been deposited at other sites). Consequently, because of the internal pressure of the protoplast, lytic death is the inevitable result of this perforation of the protective peripheral wall. Hence, the real mechanism of penicillin-induced bacteriolysis in staphylococci is considered to be mainly the result of a special morphogenetic wall defect: bacteriolysis is taking place regularly when a cell separation process is no longer preceeded by sufficient cross wall assembly at the correct place. However, hypotheses which are based purely on some variations of overall biochemical processes like total wall enzyme activities or total wall synthesis are not regarded to be sufficient to explain this type of lytic death.
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  • 31
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    Archives of microbiology 142 (1985), S. 259-261 
    ISSN: 1432-072X
    Keywords: Methanogenic bacteria ; Plasmid isolation ; Alkaline lysis ; CsCl gradient ; Restriction endonuclease mapping ; Electron microscopy ; DNA homology
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    Topics: Biology
    Notes: Abstract Of 21 recently isolated strains of methanococci, one was found to harbor a small, cryptic, low copy number plasmid. Reproducible recovery was achieved by alkaline lysis of cells pretreated with proteinase K in an osmotically stabilizing buffer. The plasmid was found to contain a singleAval site. No homology was detected between the plasmid and DNA from any of the other new strains or from five known species of methanococci.
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  • 32
    ISSN: 1432-072X
    Keywords: Immunoferritin labeling ; Electron microscopy ; Membrane vesicles ; Nitrate reductase ; Bacillus licheniformis ; Klebsiella aerogenes
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    Notes: Abstract The indirect immunoferritin labeling method was used to localize the membrane-bound respiratory nitrate reductase in membrane vesicles and protoplasts or spheroplasts of Bacillus licheniformis and Klebsiella aerogenes, respectively. For a comparison of the labeling of the various vesicle preparations, which differed not only in size but also in the percentage of inside-out orientation, a quantification of the results was needed to circumvent the problem of non-specifically bound ferritin. From the results the sidedness of the nitrate reductase in the cytoplasmic membrane of the abovementioned bacteria was determined as being cytoplasmic in B. licheniformis and as transmembranous in K. aerogenes.
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  • 33
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    Archives of microbiology 128 (1980), S. 12-18 
    ISSN: 1432-072X
    Keywords: Moraxella glucidolytica ; Electron microscopy ; Lipopolysaccharide
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    Topics: Biology
    Notes: Abstract A cellular (LPS I) and extracellular (LPS II) lipopolysaccharide were isolated from Moraxella glucidolytica cells grown on ethanol and from the culture fluid, respectively. Both LPS were toxic when injected to mice and chick embryos. These LPS contained glucose, galactose, glucosamine, galactosamine, 2-keto-3-deoxyoctonate and lipids. By permethylation studies, glucose was found to be linked (1→6) and (1→3) in LPS I and only (1→6) in LPS II. Galactose was the terminal non-reducing sugar. Branching occurred at positions 3 and 4 of galactose residues. LPS I was rich in α- and β-hydroxylauric and α-hydroxymyristic acids and LPS II contained mainly stearic and α-hydroxymyristic acids. LPS I was detoxified by mild acid and alkaline treatments. It was also dissociated by sodium deoxycholate and chromatographed on Sephadex G-75. The main fraction was reassociated by removing the surfactant by dialysis. The morphology of LPS I and LPS II was examined by electron microscopy. LPS I (original and reassociated fractions) consisted exclusively of ribbons while LPS II contained ribbons and vesicles.
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  • 34
    ISSN: 1432-072X
    Keywords: Ribulose bisphosphate carboxylase ; Quaternary structure ; Molecular weight ; Electron microscopy ; Cyanobacteria ; Synechococcus
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    Topics: Biology
    Notes: Abstract Ribulose bisphosphate (RuP2) carboxylase from the marme cyanobacterium, Synechococcus sp., comprised both large (57,000 dalton) and small (12,000 dalton) subunits. The undissociated, purified enzyme was considerably smaller than the spinach enzyme when compared by pore-gradient electrophoresis, gel filtration and density-gradient centrifugation. This suggested that the cyanobacterial enzyme might have a hexameric (L6S6) subunit structure, unlike the enzymes from spinach and many other organisms which are octamers (L8S8). However, the molecular weight of the Synechococcus enzyme was measured by equilibrium sedimentation and found to be 530,000, which is within the range observed for L8S8-type enzymes. Furthermore, electron microscopic studies of negatively stained preparations of both the native enzyme, and a preparation depleted of 87% of its small subunits by repeated mild-acid precipitation, revealed four-fold symmetry characteristic of an octameric, cubical structure. Synechococcus RuP2 carboxylase therefore must be an L8S8 octamer and its anomalous pore-penetration behaviour may be due to an asymmetric shape. Some support for the latter possibility was provided by electron miscoscopic observations of two different types of images which may be different views of the molecule in two planes.
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  • 35
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    Archives of microbiology 140 (1984), S. 265-270 
    ISSN: 1432-072X
    Keywords: EcoRI ; EcoRI-DNA complexes ; EcoRI* activity ; Recognition sites ; Frequency of binding ; Electron microscopy
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    Topics: Biology
    Notes: Abstract Electron microscopy of negatively stained isolated restriction enzyme EcoRI revealed particle projections with triangular or square outlines, indicating that the enzyme, in its tetrameric state, is tetrahedron-like. The two dimers making up the tetramer appear to be arranged in two planes orthogonal to each other. Complexes formed by EcoRI with the plasmids pBR322 or pGW10 were investigated by electron microscopic spreading techniques. In the presence of Mg2+, EcoRI was bound to the DNA molecules to form pearl necklace-like aggregates. The number of bound EcoRI particles was much higher as the sum of EcoRI-and 5′..AATT..3′ sites (with exceptions, the 5′..AATT..3′ sites may function as one type of EcoRI* sites) along the DNAs, indicating unspecific binding. In the absence of Mg2+, EcoRI was bound to the DNA only at the recognition site for EcoRI and the sites where the tetranucleotide sequence 5′..AATT..3′ was present. A direct correlation of the local concentrations of the bases A and T within the flanking sequences of the binding sites with the frequency of EcoRI to the DNA was observed. Dimers and tetramers of the enzyme was found to bind to the DNA. Tetramers occasionally exhibited two binding sites for DNA as indicated by the observation of DNA loops originating at the sites of bound tetrameric EcoRI particles.
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  • 36
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    Archives of microbiology 140 (1985), S. 338-342 
    ISSN: 1432-072X
    Keywords: Sporosarcina halophila ; Endospores ; Electron microscopy ; Heat resistance ; Ethanol resistance ; Germination ; Dipicolinic acid
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    Topics: Biology
    Notes: Abstract Sporosarcina halophila forms endospores. Electron micrographs revealed ultrastructural similarity to spores of S. ureae. Spore germination indicated by loss of refractility, darkening, swelling and formation of new vegetative cells was followed by phase contrast light microscopy. To induce spore germination, the endospores needed to be heat avtivated. After activation, they were inoculated into nutrient broth medium supplemented with sea-water. Double concentrated sea-water was found to be optimal for germination. Similar to other bacterial endospores, the spores were found to be resistant to heat and ethanol. An ultraviolet absorbing substance was isolated from suspensions of free spores; it was identified to be pyridine-2,6-dicarboxylic acid (DPA) usually present in bacterial spores. DPA was detected in amounts ranging from 5–7% of the spore dry weight; it was not detected in extracts of vegetative cells.
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  • 37
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    Archives of microbiology 141 (1985), S. 85-90 
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    Keywords: C. sporosphaeroides ; Citrate lyase ; Regulation ; Purification ; Properties ; Electron microscopy
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    Topics: Biology
    Notes: Abstract Cells of Clostridium sporosphaeroides which were grown on citrate contained citrate lyase and citrate lyase acetylating enzyme, but no detectable citrate synthase and citrate lyase deacetylase activities. Citrate lyase from C. sporosphaeroides was purified to homogeneity as judged by polyacrylamide gel electrophoresis and high performance liquid chromatography. In contrast to the enzyme from Clostridium sphenoides, the addition of l-glutamate was not necessary for activity and stabilization of the enzyme. The purified enzyme had a specific activity of 34 U/mg protein and was comparable to other citrate lyases with respect to its molecular weight and subunit composition. Electron microscopic investigations showed that similar to the lyase from C. sphenoides and in contrast to all other citrate lyases examined so far, the majority of the enzyme molecules was present in “star” form.
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  • 38
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    Archives of microbiology 143 (1986), S. 400-402 
    ISSN: 1432-072X
    Keywords: E. coli relA +/relA ; Starvation survival ; Guanosine tetraphosphate ; Electron microscopy ; Glycogen
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    Topics: Biology
    Notes: Abstract Amino acid-starved cells of Escherichia coli relA +, which contain a large number of glycogen particles, are able to survive in phosphate buffer for a longer time period than their relaxed counterparts. With regard to NH 4 + starvation differences in the survival of both strains were not found. NH 4 + starved cells of E. coli relA are able to synthesize glycogen but amino acid-starved cells of the relA strain are not. We suggest that the synthesis of glycogen triggered by guanosine tetraphosphate during amino acid starvation is responsible for the prolonged viability of the E. coli relA + strain.
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  • 39
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    Keywords: Bradyrhizobium ; Electron microscopy ; Mutants ; Nitrogen fixation ; Nodulation ; Soybean ; Symbiosis ; Transposon Tn5
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    Notes: Abstract The genome of the slow-growing Bradyrhizobium japonicum (strain 110) was mutagenized with transposon Tn5. A total of 1623 kanamycin/streptomycin resistant derivatives were screened in soybean infection tests for nodulation (Nod) and symbiotic nitrogen fixation (Fix). In this report we describe 14 strains possessing a stable, reproducible Nod+Fix- phenotype. These strains were also grown under microaerobic culture conditions to test them for free-living nitrogen fixation activity (Nif). In addition to strains having reduced Fix and Nif activities, there were also strains that had reduced symbiotic Fix activity but were Nif+ ex planta. Analysis of the genomic structure revealed that the majority of the strains had a single Tn5 insertion without any further apparent physical alteration. A few strains had additional insertions (by Tn5 or IS50), or a deletion, or had cointegrated part of the vector used for Tn5 mutagenesis. One of the insertions was found in a known nif gene (nifD) whereas all other mutations seem to affect different, hitherto unknown genes or operons. Several mutant strains had an altered nodulation phenotype, inducing numerous, small, widely distributed nodules. Light and electron microscopy revealed that most of these mutants were defective in different stages of bacteroid development and/or bacteroid persistence. The protein patterns of the mutants were inspected by two-dimensional gel electrophoresis after labelling microaerobic cultures with l-(35S)methionine. Of particular interest were mutants lacking a group of proteins the synthesis of which was known to be under oxygen control. Such strains can be regarded as potential regulatory mutants.
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  • 40
    ISSN: 1432-072X
    Keywords: Aggregation substance ; Enterococcus faecalis ; Electron microscopy ; Field emission scanning electron microscopy ; Immunogold labelling technique ; Sex pheromone system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The distribution of sex pheromone induced aggregation substance was studied on the cell surface of various Enterococcus faecalis strains. In the accompanying paper we have shown that the aggregation substance appears as a layer of hairlike structures. Using direct and indirect immunogold technique, transmission electron microscopy and high resolution scanning electron microscopy we investigated the appearance and distribution of the aggregation substance. The “hairs” increase in number with increasing exposure to sex pheromones (maximum density: 1300/μm2). We show that these structures are unequally distributed over the cell surface, even if the cells were induced by sex pheromones for a long period of time. Statistical analysis of the unequal distribution indicates that aggregation substance is incorporated into pre-existing “old” cell-walls and that this incorporation shows a saturation ca. 40 min after addition of sex pheromones.
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  • 41
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    Archives of microbiology 126 (1980), S. 87-95 
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    Keywords: Ascodichaena ; Beech bark ; Electron microscopy ; Host-fungus relationship
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    Topics: Biology
    Notes: Abstract Ascodichaena rugosa Butin is a corkinhabiting fungus, found frequently on the bark of Fagus sylvatica L. The hyphae of the fungus are distributed solely in the phellem cells, stopping their growth in the last-formed cork cell layer. The cell to cell invasion is effected by penetration hyphae, causing no extensive dissolution of the cork wall. Electron microscopical observations revealed fine structural details of the fruit bodies and of the intracellular hyphae. Of special interest were the finger-like hyaline hyphae in the last-formed layer of cork cells, which are interpreted as haustoria on the basis of the fine structure both of hyphae and host cells. This situation is considered as reflecting a parasitic relationship of Ascodichaena to beech bark. The activity of the fungus led also to the increased production of cork cells, perhaps related to the nutrient supply of the fungus.
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  • 42
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    Archives of microbiology 133 (1982), S. 97-99 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Thylakoid centers ; Photosynthetic membranes/thylakoids ; Membranes ; Membrane biogenesis ; Electron microscopy
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    Notes: Abstract An ultrastructural study of four cyanobacteria (Anabaena cylindrica, Dermocarpa violaceae, Gleocapsa alpicola, Pleurocapsa minor) indicates the presence of previously undescribed thylakoid centers from which photosynthetic membranes (thylakoids) radiate. These peripherally located thylakoid centers are cylinders 30 nm wide by 320 nm long, consisting of globular subunits oriented in nonparallel stacked arrays. Thylakoids are attached to the outer surface of the cylinder along its longitudinal axis. Thylakoid centers appear to be functionally significant due to their structure, location and thylakoid association.
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  • 43
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    Archives of microbiology 135 (1983), S. 25-29 
    ISSN: 1432-072X
    Keywords: Electron microscopy ; Euglena mutabilis ; Flagellate ; Photomovement ; Photoreceptor ; Phototaxis ; Single-cell analysis ; Videomicroscopy
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    Topics: Biology
    Notes: Abstract Due to the lack of an emergent flagellum the green flagellate Euglena mutabilis is restricted to gliding motility. During forward movement, the organisms orient positive phototactically in the presence of a suitable light stimulus. The cell contains both a stigma and a paraflagellar body which differ in shape and size from the organelles found in E. gracilis. The degree of orientation in white light follows an optimum curve with a maximum at about 100 lx. The spectral sensitivity shows a number of prominent peaks in the blue and green regions and extends well into the red region of the visible spectrum. Since the cell does not rotate during locomotion a periodic shading mechanism cannot account for phototactic orientation. Thus, phototaxis in the related species, E. gracilis and E. mutabilis differ in their photoreceptor molecules, their sensory transduction chains and their strategies of light direction detection.
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  • 44
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    Archives of microbiology 131 (1982), S. 116-123 
    ISSN: 1432-072X
    Keywords: Cell wall ; Wall degradation ; Lysozyme ; Autolysines ; Electron microscopy ; Staphylococcus aureus
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    Topics: Biology
    Notes: Abstract In contrast to former findings lysozyme was able to attack the cell walls ofStaphylococcus aureus under acid conditions. However, experiments with14C-labelled cell walls and ribonuclease indicated that, under these conditions, lysozyme acted less as an muralytic enzyme but more as an activator of pre-existing autolytic wall enzymes. Electron microscopic studies showed that under these acid conditions the cell walls were degraded by a new mechanism (i.e. “attack from the inside”). This attack on the cell wall started asymmetrically within the region of the cross wall and induced the formation of periodically arranged lytic sites between the cytoplasmic membrane and the cell wall proper. Subsequently, a gap between the cell wall and the cytoplasmic membrane resulted and large cell wall segments became detached and suspended in the medium. The sequence of lytic events corresponded to processes known to take place during wall regeneration and wall formation. In the final stage of lysozyme action at pH 5 no cell debris but “stabilized protoplasts” were to be seen without detectable alterations of the primary shape of the cells. At the same time long extended ribbon-like structures appeared outside the bacteria. The origin as well as the chemical nature of this material is discussed. Furthermore, immunological implications are considered.
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  • 45
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    Keywords: Thiobacillus ferrooxidans ; Sulfur production ; Sulfur oxidation ; Inhibitors ; Uncouplers ; Electron microscopy
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    Topics: Biology
    Notes: Abstract The intermediary production of elemental sulfur during the microbial oxidation of reduced sulfur compounds has frequently been reported. Thiobacillus ferrooxidans, an acidophilic chemolithoautotroph, was found to produce an insoluble sulfur compound, primarily elemental sulfur, during the oxidation of thiosulfate, trithionate, tetrathionate and sulfide. This was confirmed by light and electron microscopy. Sulfur was produced from sulfide by an oxidative step, while the production from tetrathionate was initiated by a hydrolytic step, probably followed by a series of chemical reactions. The oxidation of intermediary sulfur was severely inhibited by sulfhydryl-binding reagents such as N-ethylmaleimide, by the addition of uncouplers or after freezing and thawing of the cells, which probably damaged the cell membrane. The mechanisms behind these inhibitions have not yet been clarified. Finally, it was observed that elemental sulfur oxidation by whole cells depended on the medium composition. The absence of sulfate or selenate reduced the sulfur oxidation rate.
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  • 46
    ISSN: 1432-2242
    Keywords: Mitochondrial DNA of plants ; Electron microscopy ; Suspension culture ; Vicia faba
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    Notes: Summary A comparative analysis of the Vicia faba mitochondrial genome in whole plants and in longterm suspension culture has been conducted. Restriction fragment patterns of the mtDNA isolated from these two sources were notably different. Electronmicroscopic analysis also revealed significant differences. Large circular mtDNA patterns shifted from a 37–80 kb subpopulation, which was predominant in whole plants, to 18–34 kb subpopulations although in both classes notable quantities of circular molecules of 80 to 120 kb and more were also found. Both in whole plant and suspension culture cells very large circular DNAs were observed. Some of them had lengths nearly 290 kb and could be considered as evidence of the existence of master chromosomes. The minicircular DNA population was also altered. In the suspension culture we observed a notable increase of percentage of minicircles with sizes near 1 kb. Simultaneously, the percentage of minicircles with sizes near 3.5–10 kb significantly increased in suspension culture cells. In addition, a new peak (10–12 kb) of minicircles appeared. Copy number alterations for some sequences homologous to CCC1A, CCC1B and CCC2 (Negruk et al. 1982, 1985) were shown. Southern hybridization revealed the existence of a family of minicircles having sizes 1.4–2 kb with predominance of CCC1A, CCC1B and CCC2. The copy numbers of CCC1B and some minor minicircles was changed in the suspension culture when compared with the whole plants.
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  • 47
    ISSN: 1432-2242
    Keywords: Plant mitochondrial genome ; Minicircle DNA ; Electron microscopy ; Beta vulgaris L.
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    Notes: Summary The structure of mitochondrial DNA (mt-DNA) from sugarbeet (Beta vulgaris L.) has been studied by biochemical methods and electron microscopy. It was found to be complex multipartite consisting of two main classes of molecules: high molecules weight (HMW) mtDNA and low molecular weight (LMW) mtDNA. The HMW mtDNA consists of rosette-like structures and globules resembling chromomeres (150–200nm). A typical rosette has a protein core and radially stemming closed DNA loops (from 0.6-1.5 μm). The number of loops in a rosette varies from 16–30. The bulk of HMW mtDNAs are represented by interconnected rosettes (total contour length about 130–160 μm, 403–496 kbp). Such large circular DNAs may be evidence of the master chromosome arrangement of the sugarbeet genome. Globules and rosettes are interconnected by thick and thin DNA fibrils, along which nucleosome- and nucleomere-like structures are distributed. The LWM mtDNA is composed of two groups of supercoiled circular molecules, 0,2–1.5 μm and 0.02–0.05 μm in size. Electrophoretic analysis demonstrated that LWM mtDNA is represented by minicircle plasmid-like DNA molecules of 1.3, 1.4 and 1.6 kbp.
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  • 48
    ISSN: 1432-2242
    Keywords: Electron microscopy ; Genetic variation ; Mitochondrial DNA ; Oryza sativa L. ; Plasmid-like DNA
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    Topics: Biology
    Notes: Summary Mitochondrial (mt) plasmid-like DNA was found in most of more than 100 rice cultivars (Oryza sativa L.) by the use of 0.7% agarose gel electrophoresis (AGE). The DNA varied in molecular weight and number. By electron microscopy, small circular DNAs of different sizes could be detected in addition to the DNAs of high molecular weight, even in cultivars in which mt plasmid-like DNA was not detected by AGE. The detection of the mt plasmid-like DNAs by AGE did not depend on their presence or absence, but on their high stoichiometry. The relationship between cytoplasms with mt plasmid-like DNAs and varietal (for example, Indica rice) groups was close. The geographical distribution of cytoplasms is discussed.
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  • 49
    ISSN: 1432-0878
    Keywords: Smooth muscle cells ; Phenotype ; Electron microscopy ; DNA synthesis ; Man
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    Topics: Biology , Medicine
    Notes: Summary Smooth muscle cells were isolated enzymatically from adult human arteries, grown in primary culture in medium containing 10% whole blood serum, and studied by transmission electron microscopy and [3H]thymidine autoradiography. In the intact arterial wall and directly after isolation, each smooth muscle cell had a nucleus with a wide peripheral zone of condensed chromatin and a cytoplasm dominated by myofilament bundles with associated dense bodies. After 1–2 days of culture, the cells had attached to the substrate and started to spread out. At the same time, a characteristic fine-structural modification took place. It included nuclear enlargement, dispersion of the chromatin and formation of large nucleoli. Moreover, myofilament bundles disappeared and an extensive rough endoplasmic reticulum and a large Golgi complex were organized in the cytoplasm. This morphological transformation of the cells was completed in 3–4 days. It was accompanied by initiation of DNA replication and mitosis. The observations demonstrate that adult human arterial smooth muscle cells, when cultivated in vitro, pass through a phenotypic modulation of the same type as arterial smooth muscle cells from experimental animals. This modulation gives the cells morphological and functional properties resembling those of the modified smooth muscle cells found in fibroproliferative lesions of atherosclerosis. Further studies of the regulation of smooth muscle phenotype and growth may provide important clues for a better understanding of the pathogenesis of atherosclerosis.
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  • 50
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    Cell & tissue research 239 (1985), S. 219-228 
    ISSN: 1432-0878
    Keywords: Neuromuscular junctions ; Nervous system ; Electron microscopy ; Sea-urchin ; Echinodermata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The fine structure of the soft tissues at the bases of spines in the sea-urchin Echinus esculentus has been examined with particular reference to the innervation of these appendages. The basal nerve ring encircling the spine contains many somata of neurones, and circumferentially directed elements, as well as tangled neuropil. The smooth muscles that bring about spine-pointing movements are innervated by terminals that contain two different types of vesicles, suggesting dual innervation by neurones containing different neurotransmitters. The neuromuscular junctions include apparent synapses between nerve cell bodies and muscle fibres. There are also neural elements that may be involved in the control of the catch apparatus of the spine. The complexity of the nerve ring and effector innervation implies that coordination of spine movements is more sophisticated than has been previously supposed.
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  • 51
    ISSN: 1432-0878
    Keywords: Anterior pituitary gland ; Electron microscopy ; Growth hormone ; Spontaneous dwarf rats (dr/dr)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The spontaneous dwarf rat is a novel experimental model animal on the study of pituitary dwarfism. The fine structure of the anterior pituitary cells was studied in the immature and mature dwarf rats. Pituitary glands were removed from 5-, 10-, 20-day-old immature dwarfs, adult (45 days-16 weeks) dwarfs and normal 3-month-old rats and processed for electron-microscopic observation. In the control animals, growth hormone cells were readily identified by their ultrastructural characteristics, such as the presence of numerous electron-dense secretory granules, 300–350 nm in diameter, well developed rough endoplasmic reticulum and a prominent Golgi complex. In contrast, growth hormone cells were not found in the anterior pituitary gland of the spontaneous dwarf rat at any age examined. Other pituitary cell types, i.e., luteinizing hormone/ follicle stimulating hormone, thyroid stimulating hormone, adrenocorticotropic hormone and prolactin cells, appeared similar in their fine structure to those found in the control rats. In the pituitary gland of dwarf rats, a number of polygonal cells were observed either with no or relatively few secretory granules. The rough endoplasmic reticulum was arranged in parallel cisternae and the Golgi complex was generally prominent in these cells. In addition, many were found to have abundant lysosomes. A few minute secretory granules were occasionally observed; however, the immunogold technique failed to localize growth hormone or prolactin in the granules. The nature of these cells remained obscure in this study. Since their incidence and fine structural features, other than the secretory granules, were quite similar to those of the growth hormone cells in normal rats, we postulate that these cells are dysfunctional growth hormone cells. These results suggest that the cause of the growth impairment in the spontaneous dwarf rat is due to a defect in the functional growth hormone cells in the pituitary gland, and since other pituitary cell types appeared normal, the disorder seems to be analogous to the isolated growth hormone deficiency in the human.
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  • 52
    ISSN: 1432-0878
    Keywords: Antigen transfer ; Electron microscopy ; Enterocytes ; Macrophages ; Fish
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Two protein antigens, horseradish peroxidase (HRP) and ferritin, have been administered to the digestive tract of carp. Electron-microscopical observations reveal considerable absorption of both antigens in the second segment of the gut (from 70 to 95% of the total length) and also, although to a lesser extent, in the first segment (from 0 to 70% of the total length). Even when administered physiologically with food, a large amount of ferritin is absorbed by enterocytes in the second gut segment. HRP and ferritin are processed by enterocytes in different ways. HRP seems to adhere to the apical cell membrane, probably by binding to receptors, and is transported in vesicles to branched endings of lamellar infoldings of the lateral and basal cell membrane. Consequently, most of the HRP is released in the intercellular space where it contacts intra-epithelial lymphoid cells. Only small amounts of HRP become localized in secondary lysosomes of enterocytes. Ferritin does not bind to the apical cell membrane; after uptake by pinocytosis, it is present in small vesicles or vacuoles that appear to fuse with lysosome-like-bodies. In the second segment, intact ferritin ends up in the large supranuclear vacuoles (after 8 h), where it is digested slowly. Although no ferritin is found in the intercellular space, ferritin-containing macrophages are present between the epithelial cells, in the lamina propria and also to a small extent in the spleen. The transport of antigens from the intestinal lumen, through enterocytes, to intra-epithelial lymphoid cells or macrophages may have immunological implications, such as induction of a local immune response and prospectives for oral vaccination.
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  • 53
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    Cell & tissue research 239 (1985), S. 235-239 
    ISSN: 1432-0878
    Keywords: Erythropoiesis ; Autophagy ; Mitochondria ; Electron microscopy ; Stereology ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Late erythroblasts and reticulocytes from bone marrow of male Wistar rats were studied by electron-microscopic stereology. Late erythroblasts with morphological signs of nuclear extrusion (EN+erythroblasts) and late erythroblasts without these signs (EN-erythroblasts) were analysed separately. The volumes of mitochondria, autophagosomes, autophagocytosed mitochondria, autophagocytosed cytoplasm and degraded material inside autophagosomes were calculated per unit volume of cytoplasm. The results demonstrate that (1) the volume density of mitochondria in the cytoplasm decreases by 34% during maturation from (EN-)- to (EN+)-erythroblasts (p〈 0.001) and by 60% during differentiation from (EN+)-erythroblasts to reticulocytes (p〈0.001), (2) a fivefold increase in the volume density of autophagosomes in the cytoplasm is noted during maturation from (EN-)- to (EN+)-erythroblasts (p〈0.01), whereas the value of this parameter remains essentially unchanged during the subsequent differentiation to reticulocytes, (3) no mitochondria are found inside autophagosomes of (EN-)-erythroblasts, whereas mitochondria occupy 26% and 35%, respectively, of the autophagosomal volume in (EN+)-erythroblasts and in reticulocytes. Our results show that autophagocytosis of mitochondria starts at the moment of nuclear extrusion and continues in the bone marrow reticulocytes.
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  • 54
    ISSN: 1432-0878
    Keywords: Parietal cells ; Human stomach ; Peanut lectin ; Carbohydrate histochemistry ; Electron microscopy ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Peanut lectin reactivity was examined in normal fundic glands from human gastric samples, both at light- and electron-microscopic levels, using a peroxidase conjugate. Positive reaction was observed in the glycocalyx of parietal cell secretory canaliculi as well as in the mucous globules of mucous cells and in the luminal cell coat of chief cells. The presence of terminal galactose in the canalicular glycocalyx may be connected with the peculiar function of hydrochloric acid secretion. Peroxidase-labelled peanut lectin is proposed as a marker for visualizing the secretory canaliculus of parietal cells.
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  • 55
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    Cell & tissue research 239 (1985), S. 467-476 
    ISSN: 1432-0878
    Keywords: Endometrium, rat ; Stromal cells ; Hemosiderin ; Histochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cytoplasmic pigment inclusions of rat endometrial stromal cells were studied by histology, histochemistry, fluorescence microscopy, electron microscopy and X-ray microprobe analysis. It is shown that a number of endometrial perivascular stromal cells contain numerous free cytoplasmic ferritin particles as well as hemosiderin vacuoles. The larger pigment inclusions reveal also positive PAS- and Schmorl reactions indicating that they contain polysaccharide and lipofuscin material, respectively. These pigmentstoring stromal cells also display acid phosphatase activity; they avidly phagocytose instillated latex particles. No pigment-storing cells occur within the surface or glandular epithelium, either in the basal endometrium or in the myometrium. It is demonstrated that the endometrial iron-storing cells function as iron depots; they take part in the phagocytosis and endocytosis of extracellular tissue components and therefore can be named phagocytes. Our data show that “fibroblastoid” endometrial stromal cells may differentiate into endometrial resident phagocytes, which ensure interstitial proteolysis and hence facilitate the drainage of extracellular fluid into the venous blood capillaries.
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  • 56
    ISSN: 1432-0878
    Keywords: Endochondral bone development ; Osteoblast ; Osteoclast ; Horseradish peroxidase ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To investigate the mechanisms whereby bone cells absorb organic bone-matrix components during endochondral bone development, rat humeri were examined, employing horseradish peroxidase as a soluble protein tracer. Intravenously-injected peroxidase filled the osteoid layer and penetrated into the osteocyte lacunae and canaliculi, but did not enter the mineralized bone matrix. Whereas osteocytes rarely took up exogenous peroxidase, osteoblasts and osteoclasts actively endocytosed peroxidase in pinocytotic coated vesicles, tubular structures, and vacuoles. They also formed endocytotic vacuoles containing peroxidase in the Golgi area. The Golgi apparatus and dense bodies of these bone cells were, however, free of reaction products. Osteoclast ruffled borders were responsible for peroxidase absorption. In the osteoblast, osteocyte and osteoclast, endogenous peroxidatic reaction was detected only in mitochondria and not in other membrane-bounded vesicles and bodies. These results strongly suggest that both osteoblasts and osteoclasts participate in the resorption of bone-matrix organic components during bone remodelling.
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  • 57
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Secretory activity, neural control ; Transplantation ; Long-spacing collagen ; Immunocytochemistry ; Molecular markers (neuronal, glial) ; Electron microscopy ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary There is increasing evidence that, in the rat, a serotonin-mediated neural input may have an inhibitory influence on the secretory activity of the subcommissural organ (SCO). In the present investigation the rat SCO was studied 7, 30 and 90 days after transplantation under the kidney capsule, an area devoid of local serotonin-containing nerves. The grafted tissue was examined by use of immunocytochemistry employing a series of primary antisera, lectin histochemistry and transmission electron microscopy. The grafted SCO survived transplantation and contained, in addition to secretory ependymal and hypendymal SCO-cells, also elements immunoreactive with antisera against glial fibrillary acidic protein or S-100 protein. In transplants, SCO-cells produced a material displaying the characteristic immunocytochemical and lectin-binding properties of SCO-cells observed under in-situ conditions. The ependymal cells lined 1–3 small cavities, which contained secretory material. A fully developed structural equivalent of Reissner's fiber was, however, never found. The immunocytochemical and ultrastructural study of the grafted SCO showed an absence of nerve fibers within the graft and suggested a state of enhanced secretory activity. A network of protruding basal lamina structures connected the secretory cells to the newly formed capillaries revascularizing the SCO. One week after transplantation, long-spacing collagen started to appear in expanded areas of such laminar networks and also in the perivascular space. It is suggested (i) that the formation of long-spacing forms of collagen is triggered by factors provided by the SCO-secretory cells, and (ii) that secretory material of the ependymal and hypendymal cells may reach the reticular extensions of the basal lamina. In contrast to the SCO in situ, the grafted SCO-cells showed a positive immunoreaction for neuron-specific enolase. They became surrounded by a S-100-immunoreactive glial sheath that separated them from other transplanted cell types and the adjacent kidney tissue of the host.
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  • 58
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    Keywords: Proximal colon ; Distal colon ; Cholesterol ; Filipin ; Freeze-fracture ; Electron microscopy ; Guinea Pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary It is generally accepted that variations in membrane cholesterol content affect the fluidity of the bilayer, thus altering its permeability. In the biological membranes, in physiological conditions, a high cholesterol content rigidifies the bilayer decreasing its permeability, a lower cholesterol content induces the opposite effect by increasing the permeability. Since differences in the epithelial permeability for short chain fatty acids have previously been demonstrated in the proximal and distal colon of the guinea pig, these two regions were investigated to establish whether differences in membrane cholesterol content of the absorbing cells can be demonstrated. Freeze-fracture replicas of filipin-treated colonic tissue were used. The results show that in the proximal colon the density of filipin cholesterol complexes located on the luminal plasma membrane of the columnar absorbing cells was significantly higher (about twice) than in the distal colon. Therefore the lower amount of cholesterol present in the membrane of the absorbing cells in the distal colon indicates a greater fluidity of the membranes of the epithelial cells in this region. Such fluidity could be correlated to the higher absorption rates of shortchain fatty acids characteristic of this region.
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  • 59
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    Cell & tissue research 237 (1984), S. 219-226 
    ISSN: 1432-0878
    Keywords: Spermatogonia ; Spermatocytes ; Carbohydrates ; Guppy ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The structure of guppy (Poecilia reticulata) spermatogonia and spermatocytes has been studied using electron microscopy. The spermatogonia, situated at the apex of the seminiferous tubule, are almost all surrounded by a network of Sertoli cells; they have very diffuse chromatin and one or two large nucleoli. The cytoplasm contains relatively few organelles, although annulate lamellae are found. The mitochondria have few cristae and are concentrated at one pole of the cell; they are sometimes found with intermitochondrial cement. These spermatogonia are separated from each other, having no intercellular bridges or inclusion in Sertoli cells, and are relatively undifferentiated; they correspond to stem cells. The spermatogonia beneath the apex are organized into cysts. First-generation spermatogonia are more dense and heterogeneous, their nuclei becoming smaller and their chromatin becoming denser during successive generations. In spermatocytes, the synaptinemal complex exists as a modified form until metaphase. The concentration of organelles in the cytoplasm increases and the organelles become more diversified as spermatogenesis progresses. Many cytoplasmic bridges are observed (several per cell), indicating that the cells remain in contact after several divisions. These changes in germ cell structure have been related to some of the characteristic features of spermatogenesis in guppy, e.g. the large number of spermatogonial generations and the complexity of spermiogenesis.
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  • 60
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    Cell & tissue research 238 (1984), S. 413-416 
    ISSN: 1432-0878
    Keywords: Peroxisomes ; DAB-cytochemistry ; Electron microscopy ; Liver, amphibian ; Gymnophiona ; Ichthyophis glutinosus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of hepatic peroxisomes was investigated in Ichthyophis glutinosus (Amphibia: Gymnophiona), employing perfusion fixation and the diaminobenzidine (DAB) technique for the visualization of catalase. The majority of peroxisomes is circular or rod-shaped, although elongated particles occasionally occur. They contain a finely granular matrix, lightly stained after the DAB procedure. Their mean diameter is approximately 0.25 μm. Serial sections reveal that the circular and rod-shaped peroxisomal profiles are cross and oblique sections of highly tortuous, tubular organelles exceeding 2 μm in length. In addition to tubular profiles, elongated, rectangular particles, as well as straight dumbbell-shaped organelles with distinct marginal plates are observed. They range from 900 to 1650 nm in length (mean = 1200 nm). In the flattened, thin central portion of the dumbbell-shaped particle, the peroxisomal membranes form a cisterna enclosing one or two uniformly thick marginal plates, which display a definite substructure with a periodicity of 10 nm. These findings indicate that peroxisomes in the liver of Ichthyophis exhibit a complex organization. It is suggested that the organelles undergo a specific differentiation process, morphologically characterized by the formation of enlarged segments of unusual shape.
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  • 61
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    Cell & tissue research 240 (1985), S. 199-208 
    ISSN: 1432-0878
    Keywords: Ovary ; Postovulatory follicles ; Electron microscopy ; Enzyme histochemistry ; Teleosts, catfish
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In captive African catfish, Clarias gariepinus, ovulation was induced with human chorionic gonadotropin (HCG) 4 I.U./g body weight to study the function of postovulatory follicles (POFs). Ultrastructural and enzyme-histochemical data indicate that, apart from special theca cells, the granulosa of relative young POFs (i.e., from 16 h and 28 h after HCG-injection) is capable of producing steroids. Possible functions of the synthesized steroids are discussed. Histological comparison of POFs from stripped and from unstripped fish, as well as histochemical investigation of the contents of ovulated ova and granulosa of POFs at 48 h after HCG-injection, showed that the latter structure is involved in phagocytosis of the disintegrating ovulated eggs. The polysaccharide-lipid-protein material, initially taken up by heterophagolysosomes of the granulosa cells, subsequently undergoes fatty degeneration. The granulosa cells of the POFs showed strong acid phosphatase activity and abundant granular endoplasmic reticulum from 16 h after HCG-injection onward; heterophagolysosomes appeared at 32 h. These results indicate that after ovulation the phagocytotic function of the granulosa develops progressively. Autophagolysosomes, responsible for the final disintegration of POFs, become increasingly evident in the granulosa cells with increasing time after ovulation.
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  • 62
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    Cell & tissue research 240 (1985), S. 243-246 
    ISSN: 1432-0878
    Keywords: Liver ; Hepatectomy ; Hepatocyte regeneration ; Endoplasmic reticulum ; Electron microscopy ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Light-microscopic analysis of mouse liver homogenates six days after partial hepatectomy, showed a higher percentage of nuclei with adherent cytoplasm than homogenates from normal liver. This observation was true for animals with either a slow or rapid recovery of body weight after the operation. The phenomenon was not a function of the changes in the proportions of parenchymal and non-parenchymal tissue in the regenerating liver. Electron-microscopic analysis of random samples from normal and regenerating livers indicated an increase in the perinuclear rough endoplasmic reticulum, and a displacefment of the glycogen depots within the regenerating cells six days after partial hepatectomy. The marked resistance towards homogenization, shown by the cytoplasm of the regenerating cells, may have been due to the observed increase of perinuclear membranes. However, qualitative changes of the cell membranes and a general decrease of proteolytic activity connected with liver regeneration may also have contributed.
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  • 63
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    Cell & tissue research 206 (1980), S. 55-63 
    ISSN: 1432-0878
    Keywords: Phagocytosis ; Spermatozoa ; Ovum ; Fertilization ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Spermatozoa with intact acrosomes, as well as those coming into contact with the ovum at a smaller angle, and morphologically abnormal spermatozoa reach the plasma membrane of the ovum via an extensively dissolved zone of the inner layer of the vitelline membrane. This zone is assumed to be formed by overlapping of two or more tunnels formed by spermatozoa that had previously come into contact with the ovum. When a spermatozoon comes into contact with the plasma membrane of the ovum, many cytoplasmic processes extend outwards and cover it. Thereafter, the plasma membranes of the processes fuse, thereby phagocytizing the spermatozoon. It is assumed that the phagocytized spermatozoa cannot undergo transformation into male pronuclei and that they degenerate soon after phagocytosis.
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  • 64
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    Cell & tissue research 206 (1980), S. 225-232 
    ISSN: 1432-0878
    Keywords: Compound eye ; Photoreceptor membrane ; Electron microscopy ; Calcium-induced changes ; Artefacts ; Diptera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary When the compound eyes of the fly Lucilia are fixed for electron microscopy with glutaraldehyde in common buffer solutions, artefactual whorls are liable to be formed from the photoreceptor microvilli. The whorls result from two factors: (i) a prolonged time interval prior to osmication, such as the “overnight” primary fixation or wash at 4° C commonly used in studies of compound eyes; (ii) as little as 1–2 mM Ca++ in the primary fixative and wash solutions. Osmication after short (1 h) glutaraldehyde fixation at 4° C, or omission of Ca++ and addition of 2 mM EGTA, prevent whorl-formation. In the tipulid fly Ptilogyna, similar artefacts are produced, but are confined to the distal zone of the microvilli that sheds during turnover.
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  • 65
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    Cell & tissue research 211 (1980), S. 83-93 
    ISSN: 1432-0878
    Keywords: Epithelium ; Cloaca ; Electron microscopy ; Hen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The epithela of the three divisions (coprodaeum, urodaeum, proctodaeum) of the cloaca of the hen, and of the excretory ducts (colon, ureter, vagina) which join the divisions, are described using light microscopy, and scanning and transmission electron microscopy. Each region of the cloaca has its typical epithelium. Special attention is focussed in this study on the boundaries between the different epithelia. The coprodaeal epithelium does not differ considerably from that of the colon; a transitional zone is not visible. Distinct border zones, however, are observed between the other regions (ureter — urodaeum; vagina — urodaeum and proctodaeum; urodaeum-proctodaeum; proctodaeum — cutis). Although the vaginal opening is generally thought to lie in the urodaeum, our investigations show that at the vaginal opening into the cloaca the ciliated epithelium changes, on one border to a secretory epithelium characteristic of the urodaeum and on the other border to that characteristic of the proctodaeum. These observations are discussed in relation to functional aspects.
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  • 66
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    Cell & tissue research 211 (1980), S. 171-174 
    ISSN: 1432-0878
    Keywords: Pancreatic D cell ; Neural control ; Vagotomy ; Electron microscopy ; Fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In an attempt to determine the neural control of pancreatic D cells, the pancreatic islets of the domestic fowl were examined electron microscopically from 1 to 28 days after abdominal vagotomy. Exocytotic release of many secretory granules from D cells occurred one day after vagotomy. Rough endoplasmic reticulum developed and formed an arrangement of concentric whorls in the cytoplasm of D cells after axotomy. The altered D cells were also characterized by the occurrence of many peculiar dense bodies in the apical cytoplasm at all time periods studied. These bodies varied in shape and size, containing several round vesicles. The D cells were extensively depleted of granules after the longer time periods following vagotomy. The present results provide new morphological evidence for the vagus-nerve control of D cells, which may regulate the activity of islet cells.
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  • 67
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    Cell & tissue research 211 (1980), S. 191-206 
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Transplants ; Vasopressin ; Median eminence ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Blocks of anterior hypothalamus were transplanted from 19 day-old fetuses of Wistar/Lewis rats into the third ventricle of adult male Brattleboro rats. Physiological changes in graft recipients and in sham-operated animals were monitored daily. Twenty days after surgery, the graft recipients and shamoperated animals were killed and their brains examined by correlative scanning and transmission electron microscopy. Host animals that exhibited both decreased polydipsia and increased urine concentration were found to have viable grafts within the third ventricle. The observed physiological changes suggested that synthesis and release of vasopressin occurred in the transplanted neurons. Grafts were well vascularized by vessels arising from the host hypothalamus. Neurons, with perikarya ranging from 8 to 30 μm in diameter, glial cells, and neurites were located throughout the transplants. A neurohemal contact zone, similar to that normally seen in the median eminence, could not be demonstrated in the grafts. The absence of complete glial and ependymal barriers indicates a relatively close association between cells in the transplants and the cerebrospinal fluid. A large increase in supraependymal neurons and their processes, including an eruption of neurons through the floor of the third ventricle in one animal, was observed in graft recipients but not in shamoperated animals.
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  • 68
    ISSN: 1432-0878
    Keywords: Adrenergic nerves ; Acetylcholinesterase ; containing nerves ; Ovary ; Electron microscopy ; 5-Hydroxydopamine ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The autonomic nerve supply of the guinea-pig ovary was investigated by a combination of light-and electron microscopy. At the light-microscopic level, adrenergic fibres were identified due to their formaldehyde-induced fluorescence. In addition, the ovary contained acetylcholinesterase-positive fibres. In all parts of the ovary, the adrenergic fibres were most numerous. At the ultrastructural level it was possible to identify the adrenergic nerve terminals with the aid of the false adrenergic transmitter, 5-hydroxydopamine. Thus, large numbers of adrenergic terminals, characterized by their content of 50–60 nm, electron-dense synaptic vesicles, were seen within the interstitial gland, where they formed close contacts with the endocrine cells (membrane-to-membrane distance, 20–100 nm). The follicular theca externa was also richly supplied by adrenergic nerves. At this location, close contacts (50–100 nm) were identified between the nerve terminals and the smooth muscle-like cells. Very few adrenergic nerve fibres were present in the theca interna of follicles or in the corpus luteum. Non-adrenergic nerve terminals, characterized by electronlucent synaptic vesicles of 50–60 nm diameter, were observed together with the adrenergic fibres. They were always present in much lower numbers than the latter. No “p-type” nerves were identified by electron microscopy.
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  • 69
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    Cell & tissue research 242 (1985), S. 445-448 
    ISSN: 1432-0878
    Keywords: Extrahypothalamic neurosecretory neurons ; Teleost mesencephalon ; Brain stem nuclei ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neurons in the dorsal tegmentum of the midbrain of the teleosts Poecilia sphenops and P. latipinna were examined by use of electron microscopy. A nucleus of neurosecretory neurons was identified in the subependymal region just dorsal to the medial longitudinal fascicle. This nucleus has been called the dorsal tegmental magnocellular nucleus (DTMN). The most distinguishing cytological feature of these cells is the presence of large granular vesicles, 100–180 nm in diameter. These vesicles resemble neurosecretory granules characteristically found in preoptic and lateral tuberal magnocellular neurosecretory cells. Presynaptic terminals on these cells contain small clear vesicles, and some among them contain small dense-core vesicles.
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    Cell & tissue research 244 (1986), S. 385-394 
    ISSN: 1432-0878
    Keywords: Cetaceans ; Lamellar bodies ; Epidermal lipids ; Permeability barrier ; Electron microscopy ; Phocena phocena
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Biochemical and ultrastructural analysis of epidermis from the porpoise, Phocena phocena, revealed certain similarities and differences between cetaceans and terrestrial mammals. The predominant cell of cetacean epidermis, not found in normal terrestrial mammals, is a lipoker-atinocyte, which elaborates not only keratin filaments, but also two types of lipid organelles: first, lamellar bodies, morphologically identical to those of terrestrial mammals, are elaborated in great abundance in all suprabasal epidermal layers, forming intercellular lipid bilayers in the stratum corneum interstices: and second, non-membrane-bounded droplets appear and persist in all epidermal layers. Although the porpoise lipokeratinocyte morpologically resembles the sebokeratocyte of avians in certain respects, nonmembrane-bounded lipid droplets are not released into the intercorneocyte space as they are in avian stratum corneum. Whereas phospholipid/neutral lipid gradients are similar in porpoise and terrestrial mammals, PAS-positive glycoconjugates, specifically glycosphingolipids, are retained in porpoise stratum corneum, but lost from these layers in terrestrials. The novel, non-polar acylglucosyl-ceramides, which also are lost during cornification in terrestrial mammals, are retained in porpoise stratum corneum. The lipid components of porpoise lipokeratinocytes appear to subserve not only barrier function in a hypertonic milieu, but also underlie the unique buoyancy, streamlining, insulatory, and caloric properties exhibited as adaptations to the cetacean habitat.
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    Cell & tissue research 244 (1986), S. 443-448 
    ISSN: 1432-0878
    Keywords: Collagen ; Uterus ; Decidua ; Mouse ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An ultrastructural study of the features and distribution of collagen fibrils was performed in the endometrium of virgin and pregnant (2nd to 11th day) mice. Collagen-containing structures were observed in the cytoplasm of fibroblasts on the 2nd day of pregnancy. Treatment of tissues with lanthanum nitrate established that these structures were intracytoplasmic. Their association with lysosome-like bodies suggested the occurrence of intracellular digestion of collagen, probably connected with remodeling of the endometrial stroma prior to decidualization. On the 4th day of pregnancy, very few collagen fibrils were present in the intercellular space. From the 6th day of pregnancy onwards, “thick” collagen fibrils were observed between decidual cells. The diameter of these fibrils measured up to 300 nm whereas the fibrils present in the endometrium of virgin mice measured 40–68 nm.
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  • 72
    ISSN: 1432-0878
    Keywords: Stomach ; Endocrine cells ; Electron microscopy ; Morphometry ; Man
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    Notes: Summary An ultrastructural morphometric study of the endocrine cells of the oxyntic mucosa of the stomach in gastric biopsies collected from five male and five female healthy volunteers aged 19–31 was performed. No sex-related differences were disclosed. Endocrine cells accounted for 1.2±0.4% of the epithelial volume and 0.9±0.4% of the mucosal volume, i.e., including the lamina propria. After classification of the specific endocrine cell types according to the ultrastructural morphology of secretory granules, the volume densities of ECL, P and D cells (30±9%, 24±7%, and 22±4% of the entire endocrine cell mass, respectively) were higher than those of other endocrine cell types. In particular, EC cells contributed less than 10% and X cells represented a very low proportion of the total cells. Non-granulated profiles of cells which in all other respects appeared to be endocrine were also found with a volume density of 8±4%. D cells were distinguished by the high fraction of cytoplasm occupied by secretory granules (31±5%). Subdivision of the whole mucosa into four horizontal segments revealed the endocrine cells to be mostly distributed in the three lower, with virtually no endocrine cells in the superficial segment. The quantitative ultrastructural analysis of the endocrine cell population of the normal human oxyntic mucosa provided by this study may allow a better evaluation of physiological and pharmacological variations of the endocrine cell population.
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  • 73
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    Cell & tissue research 244 (1986), S. 605-612 
    ISSN: 1432-0878
    Keywords: Heart innervation ; Tissue culture ; Autonomic ganglia ; Non-neuronal cells ; Cell interrelationships ; Electron microscopy ; Guinea-pig
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    Topics: Biology , Medicine
    Notes: Summary The present study describes the ultrastructure of non-neuronal cells and their interrelationships with intracardiac neurones present in cultures dissociated atria and interatrial septum from newborn guinea-pig. When compared with the in situ preparation, most of these features in culture were similar to those observed in situ, but some differences were also apparent. Both mature and immature Schwann cells were observed in culture, and as in situ, the latter were closely associated with intracardiac neurones, whilst the former were more widely separated. The ultrastructure of satellite cells was more variable in culture than in situ: three general types were distinguished on the basis of their 10-nm filament content. This variation could be due to conditions of culture. Interstitial cells were present in culture and closely resembled those described in situ, although there was less space between cultured interstitial cells and their associated cells. Many fibroblasts, some myoblasts and a few mast cells were also found in the culture preparations.
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  • 74
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    Cell & tissue research 244 (1986), S. 595-604 
    ISSN: 1432-0878
    Keywords: Heart innervation ; Tissue culture ; Autonomic ganglia ; Neurones ; Small granule-containing cells ; Electron microscopy ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of cultured intrinsic neurones and SIF (small intensely fluorescent) cells dissociated from the atria and interatrial septum of newborn guinea-pig heart has been studied for the first time and compared with these cells in situ. Mononucleate and binucleate neuronal somata and their processes were observed in the culture preparation; their ultrastructure was similar to that of neurones in intracardiac ganglia observed in situ. The number of neurites associated with neuronal cell bodies increased after the first week in culture. A subpopulation of intracardiac neurones showed abnormalities in culture, comparable to the changes previously described in neurones of the monkey heart after unilateral vagotomy in situ. Small granule-containing cells were observed in culture, corresponding to those described in the heart in situ. One type of large process in the culture preparation containing densely packed mitochondria has not been seen in situ, suggesting that changes in cell ultrastructure due to the conditions of culture cannot be discounted. However, the ultrastructure of the cultured cells was, for the most part, consistent with that of the same cell type in situ, indicating that the culture preparation may be a useful model for investigation of the roles and interactions of intramural neurones in the heart, which are inaccessible for such studies in situ.
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  • 75
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    Cell & tissue research 246 (1986), S. 163-168 
    ISSN: 1432-0878
    Keywords: Cytoskeletal organization ; Filaments ; Triton extraction ; Replica technique ; Electron microscopy ; Amoeba proteus
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    Topics: Biology , Medicine
    Notes: Summary Various stabilization and extraction procedures were tested to demonstrate the ultrastructural organization of the cytoskeleton in normal, locomoting Amoeba proteus. Most reliable results were obtained after careful fixation in glutaraldehyde/lysine followed by prolonged extraction in a polyethylene glycol/Triton X-100 solution. Before dehydration in a graded series of ethanol and critical-point drying, the amoebae were split by the sandwich-technique, i.e., by mechanical cleavage of cells mounted between two poly-L-lysine-coated glass slides. Platinum-carbon replicas as well as thin sections prepared from such cell fragments revealed a cytoskeleton composed of at least four different types of filaments: (1) 5–7-nm filaments organized as a more or less ordered cortical network at the internal face of the plasma membrane and probably representing F-actin; (2) 10–12-nm filaments running separately or slightly aggregated through the cytoplasm and probably representing intermediate filaments; (3) 24–26-nm filaments forming a loose network and probably representing microtubules; and (4) 2–4-nm filaments as connecting elements between the other cytoskeleton constituents. Whereas microfilaments are responsible for protoplasmic streaming and other motile phenomena, the function of intermediate filaments and cytoplasmic microtubules in amoebae is still obscure.
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  • 76
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    Keywords: Penile erection ; Electron microscopy ; Immunohisto(cyto)chemistry ; Vasoactive intestinal polypeptide (VIP) ; Neuropeptide Y (NPY) ; Cercopithecus aethiops (Primates)
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    Notes: Summary The cavernous body of green monkeys contains many unmyelinated and few myelinated axons. The unmyelinated axons form terminals in the adventitia of the arteries, between trabecular muscle cells, in the interstitium, and close to endothelium cells of the sinuses. All terminals displayed predominantly “small clear vesicles” and very few “large granular vesicles”; “small granular vesicles” were not seen. However, in rabbit penises, terminals with many large granular vesicles are prominent. Immunohistochemistry (PAP technique) showed a dense network of VIP- and NPY-reactive fibres around the arteries and around trabecular muscles. The density of nerve fibres was particularly high around the subendothelial cushions of the helicine arteries. Double staining for NPY and VIP revealed that both peptides were colocalized. Immunocytochemistry (preembedding PAP technique) showed VIP- and NPY-reactivity in terminals with small clear vesicles; the reaction product was bound to the cytoplasmic face of different membrane types. Although the intracellular localization of the reaction product is probably due to artefactual displacement during preparation, the uniformity of the terminals questions the view that large and small granular vesicles in all species characterize peptidergic and noradrenergic terminals, respectively. The essential findings can be summarized as (1) a high degree of uniformity of nerve terminals, (2) colocalization of VIP and NPY, (3) heavy innervation of the subendothelial cushions of the helicine arteries, and (4) possible innervation of endothelial cells.
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  • 77
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    Cell & tissue research 256 (1989), S. 573-580 
    ISSN: 1432-0878
    Keywords: Epididymis ; Epithelium ; Monolayer culture ; Histochemistry ; Electron microscopy ; Rat (Sprague-Dawley)
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    Topics: Biology , Medicine
    Notes: Summary N-acetylglucosaminidase (NAG), acid phosphatase (ACP) and alkaline phosphatase (AKP) were localised histochemically in fixed cells from the 37-day-old rat epididymis grown in static monolayer culture for 2–8 days. ACP and NAG were cytosolic enzymes found in perinuclear positions, whereas staining of AKP was consistent with a membranous position. These enzymes were also examined in frozen tissue sections of the epididymis, from rats of the equivalent age, where NAG had intense activity in both supra- and infra-nuclear cytoplasm and ACP was more active apically. For the first time AKP was localised along basolateral membranes of the epithelium and in the lumen of the mid-caput region. The monolayer in culture was of principal cells only and they maintained their polarity and ultrastructural characteristics, but the height of the cells was reduced compared to that obtained in situ.
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  • 78
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    Cell & tissue research 257 (1989), S. 61-67 
    ISSN: 1432-0878
    Keywords: Kidney ; Peripolar cells ; Renal corpuscle ; Electron microscopy ; Raja erinacea, Mustelus canis, Rhinoptera bonasus, Sphryna lewini, Rhizoprionodon terraenovae, Squalus acanthias (Elasmobranchii)
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    Notes: Summary Granulated epithelial cells at the vascular pole of the renal corpuscle, peripolar cells, have been found in the kidneys of five species of elasmobranchs, the little skate (Raja erinaced), the smooth dogfish shark (Mustelus canis), the Atlantic sharpnose shark (Rhizoprionodon terraenovae), the scalloped hammerhead shark (Sphryna lewini), and the cow-nosed ray (Rhinoptera bonasus). In a sixth elasmobranch, the spiny dogfish shark (Squalus acanthias), the peripolar cells could not be identified among numerous other granulated epithelial cells. The peripolar cells are located at the transition between the parietal epithelium of Bowman's capsule and the visceral epithelium (podocytes) of the glomerulus, thus forming a cuff-like arrangement surrounding the hilar vessels of the renal corpuscle. These cells may have granules and/or vacuoles. Electron microscopy shows that the granules are membrane-bounded, and contain either a homogeneous material or a paracrystalline structure with a repeating period of about 18 nm. The vacuoles are electron lucent or may contain remnants of a granule. These epithelial cells lie close to the granulated cells of the glomerular afferent arteriole. They correspond to the granular peripolar cells of the mammalian, avian and amphibian kidney. The present study is the first reported occurrence of peripolar cells in a marine organism or in either bony or cartilagenous fish.
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  • 79
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    Cell & tissue research 234 (1983), S. 469-496 
    ISSN: 1432-0878
    Keywords: Bone marrow (human) ; Neutrophil granulocyte ; Granules ; Electron microscopy
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    Notes: Summary Bone marrow from hematologically healthy adults was exposed to a number of fixation procedures for investigation of the heterogeneity of the granule population in neutrophil granulocytes at the ultrastructural level. Four main cell stages were distinguished: early promyelocyte, late promyelocyte, myelocyte, and mature neutrophil granulocyte and described separately; metamyelocytes and band-form or stab cells are described together. The characteristic changes in the cytoplasm during myelopoiesis were analysed quantitatively. Special attention was given to the development of the granule population. Three types of granule arise in successive cell stages: granules which develop a sub-structure in the matrix (nucleated granules) are formed in early promyelocytes, granules with a homogeneous electron-dense matrix (azurophil granules) in late promyelocytes, and granules with a less electron-dense matrix (specific granules) in myelocytes. The three types of granule remain present during myelopoiesis. The best results in distinguishing the granule types were obtained by prefixation either in 0.1% glutaraldehyde or in 1.5% glutaraldehyde followed by washing in phosphate-buffered Ringer solution to which aminotriazole had been added. Granule counts revealed for the mature neutrophil a total number of granules of about 220 per ultrathin section. This population of granules is composed of about 12% nucleated, 11% azurophil, and 77% specific granules. When our previous findings are taken into account, the existence of three successively formed and morphologically distinguishable types of granule in heterophil (neutrophil) granulocytes has been demonstrated for three mammalian species: the guinea pig, the rat, and man. A separate term for the early promyelocyte stage is proposed: eomyelocyte.
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  • 80
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    Cell & tissue research 234 (1983), S. 579-593 
    ISSN: 1432-0878
    Keywords: Rods ; Cones ; Retina ; Bullfrog ; Synapse ; Electron microscopy
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    Topics: Biology , Medicine
    Notes: Summary Lumps of electron-dense material were observed in synaptic clefts associated with all types of photoreceptors, in the vicinity of the synaptic ribbons, in the retinae of dark-adapted frogs. Frogs were reared under a cyclic illumination (light on at 8:00; light off at 20:00) and then exposed to one of two courses of dark adaptation: one started from 11:00 in the morning, and the other started from 20:00 in the evening. The synaptic clefts of red rods became wider at some places where spherical or polygonal lumps of dense material were accumulated. The frequency and sectional area of the lumps increased faster for the first hour in the regime starting from 20:00 than in the regime starting from 11:00, then they reached the similar saturation levels of about 0.6 (per ribbon) and 1.6 to 1.8×104 (nm2) in both the regimes. In greenrod synapses, plate-shaped lumps of dense material were present in synaptic clefts and interspaces between the processes of second-order neurons. In cone synapses at the end of about 1 h darkness, the frequency and area of the lumps reached maximum values of about 0.12 (per ribbon) and 9×103 (nm2) in the regime starting from 11:00 and, about 0.08 (per ribbon) and 4 × 103 (nm2) in the regime starting from 20:00. On exposure to light, the dense material abruptly disappeared from all types of photoreceptor synaptic clefts. Large dense-core vesicles, occasionally observed in light-adapted rod photoreceptor terminals, seem to participate in exocytosis of the dense material. The number of dense-core vesicles per synaptic ribbon in a terminal was about 0.55 at the end of 3 h light in the morning and about 1.28 at the end of 12 h light in the evening. The increased number of dense-core vesicles during the daytime may contribute to the faster accumulation of dense material in the synaptic clefts. Although the chemical identification or the functional significance of the electron-dense material remains unknown, it is interesting that this material showed a rise and fall in response to darkness and illumination. Also the fact that this material is clearly visible will be helpful for future analysis of frog photoreceptor synapses.
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  • 81
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    Keywords: Ovulation (rabbit) ; Graafian follicle ; Perfusion ; Electron microscopy
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    Notes: Summary Isolated ovaries from untreated, sexually mature rabbits were introduced into an in vitro perfusion system and perfused with a chemically defined medium containing albumin. The ovaries were perfused for up to 15 h (mean 11.5 h) and then processed for morphological investigation. Both at the light- and electron-microscopical levels, most of the ovaries exhibited a normal structure comparable with ovaries in situ. In two cases, however, marked accumulations of bacteria were found, although not inside the follicles. Since ovulation in the rabbit normally occurs between 9.5–13 h after mating or human chorionic gonadotrophin treatment, this model seems adequate for studies of ovulation in vitro. It is, however, important to study the ovaries microscopically after the perfusion to detect artifacts, e.g., bacterial infection, that may have influence on the process of ovulation.
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  • 82
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    Keywords: Tannic acid ; Acetylcholine receptors ; Tissue culture ; Electron microscopy
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    Notes: Summary Spinal cord neurons from 9-day chick embryos were maintained in culture for up to 35 days and then fixed in 4% cacodylate-buffered glutaraldehyde containing 2% tannic acid. After about 15 days in culture a small percentage of the synaptic specializations present were characterized by striking electron-dense striations averaging 15 nm in width, oriented perpendicular to the postsynaptic membrane. These structures increased in frequency with time in culture (to a maximum of about 10% of all synapses in the oldest cultures); they were asymmetrical, protruding approximately 8 nm into the synaptic cleft, and more deeply (approximately 15–18 nm), into the postsynaptic cytoplasm. On the basis of earlier work by Sealock (1980) they are interpreted as concentrations of acetylcholine receptors. Similar membrane differentiations were also seen associated with active-zone areas of a few presynaptic membranes, and the possibility that these represent presynaptic acetylcholine receptors is discussed. Additional observations reported are (1) the presence of striations resembling those seen at the postsynaptic membrane in the membranes of some postsynaptic vesicles, and (2) filamentous links between the striations and cytoskeletal elements of the postsynaptic cell.
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  • 83
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    Keywords: Monoamine-containing cells ; Taste bud ; Paracrine cells ; Mechanoreceptors ; Electron microscopy ; Teleosts
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    Notes: Summary The taste buds on the barbels in three species of teleosts (Cyprinus carpio, Misgurnus anguillicaudatus, Parasilurus asotus) were studied by means of fluorescence and electron microscopy. Intensely yellow-fluorescent cells, which are disk-shaped and located exclusively in a basal position, are observed in the barbel-buds of all fishes examined. The basal cells contain a large number of small clear vesicles approximately 40–60 nm in diameter, which show a tendency to aggregate in the cytoplasm facing the junction of the nerve terminals; chemically transmitting synapses are seen in the latter region. It is suggested from the present observations that the basal cells in the barbel-bud may originate from Schwann cells and have a dual function both as mechanoreceptors and paracrine elements. Since the administration of 5,6-DHT results in an appearance of small dense vesicles among the small clear vesicles, the possibility exists that the basal cell may be capable of taking up monoamines and storing them in the small clear vesicles.
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  • 84
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    Keywords: Exocrine pancreas ; Calcium pool ; Calcium release ; Electron microscopy ; Rat
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    Notes: Summary In an attempt to identify a cellular Ca2+-pool, from which calcium is released when secretagogues are applied, tissue fragments of the rat exocrine pancreas were incubated and fixed with glutaraldehyde in the presence of calcium. By means of this procedure electron-dense deposits were found on plasma membranes. X-ray microanalysis showed that these deposits contain calcium. Stimulation of tissue fragments with the use of the secretagogues carbachol or cholecystokinin reduced the number of deposits by about 80%. When the antagonist atropine was applied after carbachol stimulation, deposits reappeared on cell membranes, which then disappeared again after a second stimulation with cholecystokinin. In the presence of procaine, carbachol was inhibited and only slightly reduced the Ca2+-deposits on the plasma membranes. These results suggest that a calcium pool, from which calcium is released to induce enzyme secretion on stimulation, is located in the cell membrane
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  • 85
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    Keywords: Chloride cells ; Acid stress ; Gill ; Electron microscopy ; Fathead minnow
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    Notes: Summary Fathead minnows, Pimephales promelas, were exposed for 129 days to Lake Superior water acidified with sulfuric acid by means of a flow-through toxicant injection system. The effects of chronic acid stress (pH 6.5, 6.0, 5.5, 5.0) on gill histology were examined. Most of the histological effects were seen at pH 5.5 and 5.0 and were confined primarily to changes in numbers, distribution, and morphology of chloride cells. At low pH levels there tend to be more chloride cells in the gill epithelium and an increased percentage of these cells in the secondary lamellae. In contrast to normal chloride cells, chloride cells from fish exposed to low pH frequently had apical pits while some had bulbous apical evaginations. The occurrence of structural changes in chloride cells during exposure to acid water suggests that chloride cells may be involved in acclimation to acid stress.
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  • 86
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    Keywords: Lactating cell ; Lipid droplets ; Secretory vesicles ; Mitochondria ; Intracellular associations ; Electron microscopy ; Milk secretion
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    Notes: Summary The size, cellular location, and identity of surface-associated components were determined for lipid droplets in lactating cells. Transmission electron-microscopic measurements were made involving 3801 droplets in approximately 211 cells from three rats and 1197 droplets in 66 cells from a mouse. For the purposes of droplet evaluation, cells were divided into seven locations ranging from basal to secreting positions. Droplets were also categorized with respect to contact with other droplets, basolateral plasma membrane, mitochondria, Golgi apparatus, secretory vesicles, and endoplasmic reticulum-cytoplasm (ERC). Data on droplet size showed that droplet growth occurs mainly in the secretory position, confirming previously published findings. Lipid droplets from mouse tissue, although somewhat smaller in size showed similar growth trends to those of the rat. Data on numbers of droplet contacts and percentages of droplet circumferences involved in associations with other cell components showed that the dominant interaction of lipid droplets was with the ERC. However, intimate association of droplets with mitochondria was noted in all cellular locations. In addition, nursed animals exhibited a greater proportion of droplet surface association with secretory vesicles and less in contact with mitochondria in comparison to those not nursed. The significance of these relationships to milk synthesis and secretion is discussed.
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  • 87
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    Cell & tissue research 236 (1984), S. 249-255 
    ISSN: 1432-0878
    Keywords: Oocyte ; Nucleolus ; Silver staining ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The nucleoli of dictyate-stage growing oocytes in rat ovaries were examined both with routine electron microscopy and electron microscopy after silver nitrate and ammoniacal silver nitrate (Ag-AS) staining. The nucleoli of the unilaminar follicular oocytes consist of twisted strands of dense fibrillar components, aggregates of granular components, and small fibrillar centers. After Ag-AS staining, silver grains are numerous on the dense fibrillar strands, fewer on the fibrillar centers, and very sporadic on the granular aggregates. The same stainability of three nucleolar components with the Ag-AS method was also confirmed in the nucleoli segregated by actinomycin D. During the transition of growing oocytes from bilaminar to plurilaminar follicle stage, the nucleolar dense fibrillar strands gradually conglomerate and are transformed into large and compact spherules. The stainability of dense fibrillar components with the Ag-AS method was lost along with this nucleolar transformation. These results may provide some new clues on the functional significance of AgAS-positive proteins in the nucleoli.
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  • 88
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    Cell & tissue research 206 (1980), S. 303-318 
    ISSN: 1432-0878
    Keywords: Area postrema, rat ; Ependyma ; Cyst ; Circumventricular organs ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Peculiar cells forming cysts were observed in the area postrema and sometimes also in the choroid plexus and the tela chorioidea near the area postrema, and were studied in detail by electron microscopy. The cytological features of the cyst cell and its junctional relationship to neighboring cells imply that cyst cells are derived from ependymal and choroid epithelial cells. The cyst cells usually contact directly the perivascular spaces of postremal, choroidal or pial capillaries, where the cytoplasm is often considerably attenuated. The cystic lumen is commonly filled with a flocculent material. The limiting membrane of the cystic lumen, which frequently bears cilia and microvilli, has the same thickness as the surface cell membrane. In many cases, the cyst is surrounded by the cytoplasm of a single cell. In some cases, however, two cells participate in the formation of the cyst, although one is only a slender process and joined by a zonula occludens with the main cyst cell. Horseradish peroxidase (HRP) injected into the cerebrospinal fluid (CSF) space failed to enter the cystic lumen. A possible significance of the cyst in relation to the CSF and blood circulation was considered.
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  • 89
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    Keywords: Vocal motor system ; Songbirds (zebra finch) ; Calcium-binding proteins ; Parvalbumin ; Electron microscopy ; Plasticity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of parvalbumin (PV) within neurons of the vocal motor nucleus hyperstriatum ventralepars caudalis (HVc) was investigated in the forebrain of adult male zebra finches by means of light and electron microscopy using the indirect immunoperoxidase technique. Parvalbumin-reaction product was located in the amorphous material of perikarya, dendrites and nuclei, and associated to microtubuli, postsynaptic densities and intracellular membranes; it was found in some axons and Gray type-2 boutons, but rarely in type-1 boutons and never in the Golgi apparatus. These observations suggest that parvalbumin may regulate calcium-dependent processes at the postsynaptic membrane and in the cytosol. Furthermore, the partial association of parvalbumin to microtubuli points to an involvement in calcium-dependent tubular functions. Calcium currents and microtubular assembly or transport may be relevant for the known functions of HVc in song learning.
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  • 90
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    Cell & tissue research 210 (1980), S. 447-459 
    ISSN: 1432-0878
    Keywords: Lipid ; Kidney tubules, proximal ; Autoradiography ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Lipid metabolism in the cells of the renal proximal convoluted tubules (PCT) was investigated in healthy fowls and in fowls with the Fatty Liver and Kidney Syndrome (FLKS). The tissue was fixed at 10–25 min intervals after intravenous injection of 3H-oleic acid. The distribution of autoradiographic grains was analysed by the “circle method”. In normal cells most of the silver grains were associated with the cytoplasmic organelles. Lipid droplets and Golgi elements had the highest specific activity relative to the nuclear activity, which was little above background level. Lysosome-like bodies and mitochondria had lower values. In the cells of the FLKS-affected birds a large proportion of the grains was located over the lipid droplets, which are abundant in this condition. The specific activity of the cytoplasmic organelles was barely 2-fold higher than the nuclear activity. The results suggest that there is a diminished incorporation of esterified fatty acids by the organelles of these cells and that the excess is transferred to the lipid droplets. The identity of low electron density particles observed in the PCT cells of severely affected birds is discussed.
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  • 91
    ISSN: 1432-0878
    Keywords: CRF immunohistology ; Paraventricular nuclei ; Pituitary gland, pars nervosa ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary A fine network of corticotropin-releasing factor (CRF)-immunopositive fibers was found in the posterior lobe of the pituitary of the rat. The intermediate and distal lobes were free of CRF-immunoreactivity. Varicose, terminal-like axons were frequently observed around capillary vessels. Surgical isolation of the paraventricular nuclei resulted in a complete disappearance of CRF-immunoreactive fibers from the posterior lobe. CRF-immunopositive fibers show the general characteristics of peptidergic axons. These ultrastructural observations support the idea that CRF is secreted into capillary vessels.
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  • 92
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    Cell & tissue research 211 (1980), S. 331-343 
    ISSN: 1432-0878
    Keywords: Gut hormones ; Endocrine cells ; Electron microscopy ; Immunocytochemistry ; Peptidergic innervation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Histological, cytochemical and immunocytochemical methods were used in light and electron microscopical studies to demonstrate the presence of a neuroendocrine system in the gut of the urodele, Salamandra salamandra. Cytochemical stains capable of detecting peptide-producing endocrine cells demonstrate cells reacting with Masson's silver (argentaffin) method, Grimelius' argyrophil silver method, masked metachromasia method and the lead haematoxylin stain. Using antisera raised to a variety of mammalian gut peptides, cells containing bombesin-, gastrin-, somatostatin-, substance P- and glucagon-like immunoreactivity were identified; vasoactive intestinal polypeptide- and substance P-like immunoreactivities were found in nerve fibres in the submucous and myenteric plexus. No immunoreactivity was detected for motilin, gastric inhibitory polypeptide, cholecystokinin or secretin. The ultrastructure of the immunoreactive cells and nerves was revealed by the semithin/thin method. All the cells identified contained numerous electrondense secretory granules, which varied in their chracteristic morphological structure from one cell type to another. The evidence collected in this study indicates that a complex neuroendocrine system regulating gut function is present in this amphibian and may have developed prior to the emergence of the phylum.
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  • 93
    ISSN: 1432-0878
    Keywords: Mammary gland ; Tissue culture ; Collagen gel ; Electron microscopy ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Fragments of human breast epithelium, devoid of all stromal and basal lamina components, which maintain their in vivo topological organisation can be cultured for up to 28 days within a reconstituted rat-tail-derived collagen matrix. These organoids initially undergo a loss of structural and 3-dimensional organisation, typified by loss of lumina formed by epithelial cells, and myosin from myoepithelial cells. Their subsequent reorganisation is dependent on the presence of serum, insulin, hydrocortisone, and cholera toxin in tissue culture medium. After this preliminary phase, a reduction in the concentration of serum, insulin, hydrocortisone, and cholera toxin is necessary to allow the structural differentiation of epithelial and myoepithelial cells. The myoepithelial cells also regain their ability to produce the basal lamina component laminin. The use of bovine-dermal collagen as the matrix, rather than rat-tail-derived collagen is shown to result in more stable organisation and differentiation of the organoids. The successful use of single-cell pellets (derived by trypsinisation of the organoids) in place of organoids in such cultures illustrates that there is no requirement for pre-existing cell/ cell contact or topological organisation of cells prior to embedding within the collagen matrix.
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  • 94
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    Cell & tissue research 247 (1987), S. 457-459 
    ISSN: 1432-0878
    Keywords: Vomeronasal glands ; Autonomic innervation ; Electron microscopy ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphological evidence for a direct autonomic innervation of the mouse vomeronasal glands is presented. Axonal varicosities containing a few densecore vesicles and numerous clear vesicles (36–60 nm in diameter) make synaptic contacts with the secretory cells at the base of the glandular acini. The axonal presynaptic membrane is associated with a distinct dense material and it is separated from the secretory cell by a synaptic cleft of about 12–14 nm. At the postsynaptical level, coated vesicles can be found. Additional postsynaptical specializations have not been observed.
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  • 95
    ISSN: 1432-0878
    Keywords: Thyroid follicle ; Suspension culture ; Cellular polarity ; Tight junctions ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Porcine thyroid follicles, when isolated by enzymatic digestion and suspended in Eagle's MEM containing 10% fetal calf serum, undergo inversion of cellular polarity. After isolation, the strands for the tight junctions (zonulae occludentes) between follicle cells begin to move towards the side of the medium and gather at this side of the lateral plasma membrane during 24 h of incubation. Around this time, microvilli of many follicular cells protrude into the culture-medium. The elements of the Golgi apparatus are located at the luminal as well as the culture-medium side of the cytoplasm, and also at the lateral side of the nucleus after 24 h of suspension culture, and by 94 h of incubation almost all elements of this organelle, as well as lysosomes and the central cilium have migrated to the side of the medium. The migration of the zonulae occludentes is considered to be the initial change in the reversal of the polarity of this cell.
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  • 96
    Electronic Resource
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    Cell & tissue research 248 (1987), S. 49-54 
    ISSN: 1432-0878
    Keywords: Enterochromaffin cells ; Serotonin ; Duodenum ; Immunocytochemistry ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Enterochromaffin cells of the rat duodenum have been studied immunocytochemically by use of a specific antiserum to serotonin. At the light-microscopic level serotonin immunoreactivity was observed in enterochromaffin cells located in the epithelium of the duodenal mucosa. Most of the serotonin-immunoreactive material was localized to the basal portion of the enterochromaffin cells, but small amounts of immunoreactive material were regularly observed in the apical portion. At the electron-microscopic level serotonin immunoreactivity in enterochromaffin cells was found to be concentrated over the dense cores of the cytoplasmic granules. The majority of these granules was located in the basal cytoplasm of the enterochromaffin cells, but serotonin-immunoreactive granules were also observed in the apical cytoplasm immediately beneath the microvilli. These observations indicate that duodenal enterochromaffin cells are bipolar and that they secrete serotonin both basally, to the circulation, and apically, to the gut lumen. Rat duodenal enterochromaffin cells thus appear to have an exocrine as well as an endocrine function.
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  • 97
    Electronic Resource
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    Springer
    Cell & tissue research 248 (1987), S. 675-682 
    ISSN: 1432-0878
    Keywords: Molluscan muscles ; Innervation ; Chromatophores ; Cephalopods ; Electron microscopy ; Eledone cirrhosa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cephalopod chromatophores are made of a central pigment cell surrounded by 10 to 20 radially arranged muscle fibres under direct nervous control. Innervation of these muscle fibres was studied with anterograde cobalt fills of peripheral nerve bundles and light and electron microscopy. Individual axons branch repeatedly to innervate the muscles of chromatophores scattered over several millimeters. Axons contained in several dermal nerves converge to innervate the same chromatophores. Among the chromaophores, axons were found running either singly or in small bundles, often accompanied by sheath cells. Single chromatophore muscles were innervated by at least one axon running across or along its length. Since nerves terminating on chromatophore muscles are very rare, neuromuscular contact seems to be made “en passant”. Varicosities of the axons apposed to the muscles are thought to be presynaptic sites. However, morphological differentiations of the pre-or post-synaptic membranes were not visible. Two types of innervating processes were found containing either electron-clear or a mixture of electron-clear and dark-core synaptic vesicles.
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  • 98
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    Cell & tissue research 254 (1988), S. 341-346 
    ISSN: 1432-0878
    Keywords: Odontoblast ; Calbindin ; Immunohistochemistry ; Electron microscopy ; Teeth ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence of 28 kDa calbindin in human odontoblasts was studied by use of specific antibodies raised against chick duodenal 28 kDa calbindin, in immunofluorescence, immuno-peroxidase, and electron-microscopic labelling experiments. The calbindin-like protein was detected mainly in the cytoplasm of odontoblast cell bodies, in their processes and occasionally in their nuclei. Correspondingly, at the ultrastructural level, immunoreactive material was associated with the cytosol, microfilaments and cilia. These findings suggest that human odontoblasts express a 28 kDa vitamin D-dependent calcium-binding protein, unlike those of rats and mice in which ameloblasts are the only cells immunoreactive for the protein.
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  • 99
    Electronic Resource
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    Springer
    Cell & tissue research 254 (1988), S. 449-454 
    ISSN: 1432-0878
    Keywords: Follicular dendritic cells ; Lymph nodes ; Electron microscopy ; Nude mice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We have studied follicular dendritic cells (FDC) in lymph nodes of normal and thymus dysgeneic nude mice depleted of B-cells by chronic treatment with anti-IgM antibodies. We found that B cell depletion was accompanied by the absence of mature FDC as defined morphologically at the ultrastructural level. Only precursor FDC (p-FDC) could be demonstrated. Upon release of B-cell suppression, the repopulation of lymph nodes with B-cells was associated with the reappearance of fully differentiated FDC in primary follicles of nude mice and in secondary follicles of T-cell competent mice. We conclude that mature B-cells and/or B-cell products are required for the development of mature follicular dendritic cells in the mouse lymph node.
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  • 100
    ISSN: 1432-0878
    Keywords: Electron microscopy ; Malaria parasites ; Merozoites ; Surface coat ; Maturation ; Plasmodium knowlesi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The surface of extracellular merozoites of P. knowlesi is covered with a coat 15–20 nm thick, made up of clusters of filaments standing erect on the plasma membrane. Filaments have stems 2 nm thick, the peripheral ends of which are complex, branching or ending in long trailing threads. Coat filaments occur on the surface of the parasite in regular rows at an early schizont stage, and persist until well after merozoite release. They are sensitive to trypsin and papain, and bind ethanolic phosphotungstate, indicating a proteinaceous nature. They are also removed by exposure to phosphate-buffered saline. Filaments bear negative charges, binding cationised ferritin throughout the depth of the coat and staining with ruthenium red. They cover the whole merozoite surface and mediate intercellular adhesion at distances of 15–150 nm, membrane to membrane. It is suggested that these filaments correspond to a major merozoite surface protein, and are important in the initial capture of red cells.
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