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  • Electron microscopy  (1,196)
  • Rat  (826)
  • Springer  (1,967)
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  • 1
    ISSN: 1432-041X
    Keywords: Inner cell mass ; Blastocyst ; Morula ; Implantation ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The number of trophectoderm (TE) and inner cell mass (ICM) cells was determined by complementmediated lysis and differential staining in rat embryos collected at different times during in vivo preimplantation development. At 90 h after fertilization, two groups of morulae were discriminated according to the presence or absence of detectable ICM cells, and the analysis of their total cell number indicated that acquisition of a permeability seal between TE cells begins at the 14-cell stage. On the other hand, our data confirmed that blastocoele formation occurs after the fourth cleavage division in the rat. The total cell number increased exponentially with time in blastocysts recovered between 90 h and 127 h but the cell kinetics of TE and ICM cells were different. The proportion of ICM cells consequently varied throughout blastocyst development, with a peak value for expanded blastocysts at 103 h. Finally, a linear-quadratic relationship was found between the numbers of TE and ICM cells when all the embryos with a detectable ICM were analysed together.
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  • 2
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    Development genes and evolution 206 (1997), S. 503-514 
    ISSN: 1432-041X
    Keywords: Key words Preimplantation mouse embryo ; Brefeldin-A ; Monensin ; Golgi ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The intracellular trafficking of integral membrane and secreted proteins is likely to be a key element involved in the morphogenesis and differentiation of the early mammalian embryo. In this study, we used transmission electron microscopy (TEM) to analyse the effects of brefeldin-A (BFA) and monensin, well known inhibitors of vesicular protein trafficking in somatic cells, on the structure of preimplantation mouse embryos. Both BFA and monensin distinctively altered the morphology of Golgi compartments in the blastomeres of treated morulae. BFA-treated morulae lacked recognizable Golgi complexes but possessed heterogeneous organelle clusters consisting of an abundance of smooth tubular and vesicular membrane compartments in addition to mitochondria, endosomes and lysosomes. Treatment of morulae with monensin was associated with swelling of Golgi compartments in addition to altering the morphology of mitochondria, lysosomes and the plasma membrane. BFA, and to a lesser extent monensin, inhibited cytokinesis as evidenced by the detection of binucleate blastomeres. In addition, BFA induced morulae to decompact. These latter effects have not been reported previously for these agents in mammalian somatic cell lines or other vertebrate or invertebrate embryos. These results provide the first demonstration of the structural effects of BFA and monensin on cells of the early mammalian embryo, some of which are consistent with the known actions of these agents on components of the vesicular protein trafficking system in mammalian somatic cells. This information serves as a foundation for the further use of these agents in studies of vesicular protein trafficking as an agent of preimplantation morphogenesis.
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  • 3
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    Development genes and evolution 194 (1985), S. 429-432 
    ISSN: 1432-041X
    Keywords: Tail bud ; Tail gut ; Gut ; Organogenesis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The formation of the tail portion of the primitive gut was investigated by light and electron microscopy in 10- and 11-day rat embryos. The observations permit the conclusion that the tail gut does not form as a posterior extension of the hindgut but originates from the tail bud mesenchyme by mechanism analogous to the secondary neurulation. It includes cell condensation, aquisition of apicobasal polarity and the radial, rosette-like arrangement around a central cavity. These cells bear the cytological characteristics of both the absorptive epithelial cells and the mesenchymal cells at their apical (adluminal) and abluminal ends respectively.
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  • 4
    ISSN: 1432-041X
    Keywords: Synaptogenesis ; Electron microscopy ; Visual acuity ; Fish development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The morphogenetic differentiation of synapses of the optic tectum of the rainbow trout was investigated at different stages of development (from hatching to adult) and compared with the improvement in visual discrimination (minimum separable). (1) The main phase of synaptogenesis (increase in number of synapses, length of contact zone and number of vesicles) begins about one week after hatching and continues up to the age of one month, when the larvae start swimming freely. (2) Myelination begins 26 days after hatching and induces the end of the synaptogenesis period. (3) The visual discrimination (minimum separable) of trout larvae improves from 30 degrees of arc on the 10th day after hatching to 1 degree on day 30, then to about 14 to 18 min of arc in the adult. The results are discussed with special reference to previous biochemical investigations on changes in the ganglioside composition of the trout brain during comparable periods of development.
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  • 5
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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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  • 6
    ISSN: 1420-9071
    Keywords: Rat ; alloxan diabetes ; intestinal villus ; alkaline phosphatase ; sucrase ; epithelial cells ; villus-crypt axis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The sucrase activity in enterocytes isolated from the villus crypt axis was found to increase in all regions of the villus from day 2 after induction of diabetes, and the increase continued until day 4. In contrast, alkaline phosphatase activity increased mainly in the apical one-third of the villus-crypt column, and the increase occurred abruptly on day 4 with increase in food intake.
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  • 7
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    Cellular and molecular life sciences 42 (1986), S. 169-171 
    ISSN: 1420-9071
    Keywords: Rat ; olfactory pathway selective lesions ; plasma corticosterone ; lactacidemia ; adrenal glands
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In comparison to control rats, basal plasma corticosterone level and lactacidemia significantly increased in rats submitted to a bilateral lesion of the lateral olfactory tract and/or the anterior branch of the anterior commissure. Only the anterior branch of the anterior commissure induced hyperglycemia; that of the lateral olfactory tract exerted an opposite effect.
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  • 8
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    Machine vision and applications 4 (1991), S. 271-285 
    ISSN: 1432-1769
    Keywords: Electron microscopy ; three-dimensional vision ; surface reconstruction ; stereo ; shape from shading ; dynamic programming
    Source: Springer Online Journal Archives 1860-2000
    Topics: Computer Science
    Notes: Abstract The computational reconstruction of surface topographies from scanning electron microscope (SEM) images has been extensively investigated in the past, but fundamental image processing problems still exist. Since conventional approaches adapted from general-purpose image processing have not sufficiently met the requirements in terms of resolution and reliability, we have explored combining different methods to obtain better results. This paper presents a least-squares combination of conventional stereoscopy with “shape from shading” and a way of obtaining self-consistent surface profiles from stereoscopy and “stereo-intrinsic shape from shading” using dynamic programming techniques. Results are presented showing how this combined analysis of multi-sensorial data yields improvements of the reconstructed surface topography that cannot be obtained from individual sensor signals alone.
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  • 9
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    The journal of membrane biology 140 (1994), S. 215-223 
    ISSN: 1432-1424
    Keywords: Insulin receptor ; Membrane reconstitution ; Electron microscopy ; Quaternary structure ; Immunogold labeling
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Insulin receptors were incorporated into liposomes by two different procedures, one using dialysis and one using detergent removal by Bio-Beads. Receptor incorporation was analyzed by gradient centrifugation and electron microscopy. Reconstituted receptors projected up to 12 nm above the membrane and exhibited a T-shaped structure compatible with that previously described for the solubilized receptor. Insulin binding and autophosphorylation experiments indicated that approx. 50% of the receptors were incorporated right-side out. Such random orientation was confirmed by immunogold labeling of the α- and the β-subunit of the receptor. Immunogold labeling of the C-terminus of the β-subunit indicates that it resides about 6 nm off the membrane, while two α-subunit epitopes were labeled at about twice this distance, confirming that the α-subunit is harbored in the cross-bar of the T-structure.
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  • 10
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    Journal of molecular evolution 24 (1987), S. 260-271 
    ISSN: 1432-1432
    Keywords: Mouse ; Rat ; Two-dimensional electrophoresis ; Quantitative variability of proteins ; Qualitative variability of proteins ; Protein classes ; Membrane proteins ; Organ-specific proteins ; Regulatory genes ; Speciation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Proteins of membranes and cytosols were extracted from the livers and brains of mice (inbred strain DBA/6J) and rats (inbred strain DA/Han) and separated by two-dimensional electrophoresis (2-DE). The 2-DE patterns were compared with regard to qualitative (spot position) and quantitative (spot intensity) characteristics of the proteins of these two species. The following results were obtained: (1) Brain had more (higher percentage) conservative proteins (proteins found in both mice and rats) than liver; (2) plasma membranes had more conservative proteins than the cytosols; (3) organ-unspecific proteins contained more conservative proteins than relatively organ-specific proteins; (4) the pattern of distribution of genetic variability among different classes of proteins represented by findings 1–3 was the same for the qualitative and quantative characteristics of the proteins; and (5) some observations indicated that quantitative variability occurred more frequently among proteins than did qualitative variability. Our conclusion is that regulatory sequences in the DNA (regulatory genes) are subjected to functional constraints that differ in strength among different classes of proteins by the same ratios as the constraints acting on the structural genes. The overall effect of the selective pressure is, however, less stringent for regulatory genes than for structural genes. The results obtained here by comparing two different species are very similar to previous results we obtained by studying different subspecies (inbred strains of the mouse). From this finding arises a new concept: the study of molecular evolution on the basis of different classes of proteins. Our results were compared with data from the literature that were obtained in part from studies on cultured cells. The comparison suggested that cultured cells have lost their tissue-specific proteins, and so generate predominantly extremely conservative proteins.
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  • 11
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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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  • 12
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    Journal of molecular evolution 33 (1991), S. 442-449 
    ISSN: 1432-1432
    Keywords: Humans ; Mouse ; Rat ; Codon usage ; Mutation bias ; Selection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A new statistical test has been developed to detect selection on silent sites. This test compares the codon usage within a gene and thus does not require knowledge of which genes are under the greatest selection, that there exist common trends in codon usage across genes, or that genes have the same mutation pattern. It also controls for mutational biases that might be introduced by the adjacent bases. The test was applied to 62 mammalian sequences, the significant codon usage biases were detected in all three species examined (humans, rats, and mice). However, these biases appear not to be the consequence of selection, but of the first base pair in the codon influencing the mutation pattern at the third position.
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  • 13
    ISSN: 1432-1432
    Keywords: Carcinoembryonic antigen ; Evolution ; Gene family ; Human ; Rat ; Synonymous substitutions ; Silent molecular clock ; Evolutionary trees
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Various rodent and primate DNAs exhibit a stronger intra- than interspecies cross-hybridization with probes derived from the N-terminal domain exons of human and rat carcinoembryonic antigen (CEA)-like genes. Southern analyses also reveal that the human and rat CEA gene families are of similar complexity. We counted at least 10 different genes per human haploid genome. In the rat, approximately seven to nine different N-terminal domain exons that presumably represent different genes appear to be present. We were able to assign the corresponding genomic restriction endonuclease fragments to already isolated CEA gene family members of both human and rat. Highly similar subgroups, as found within the human CEA gene family, seem to be absent from the rat genome. Hybridization with an intron probe from the human nonspecific cross-reacting antigen (NCA) gene and analysis of DNA sequence data indicate the conservation of noncoding regions among CEA-like genes within primates, implicating that whole gene units may have been duplicated. With the help of a computer program and by calculating the rate of synonymous substitutions, evolutionary trees have been derived. From this, we propose that an independent parallel evolution, leading to different CEA gene families, must have taken place in, at least, the primate and rodent orders.
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  • 14
    ISSN: 1432-1211
    Keywords: Key words CD1 ; Rat ; Gene ; Organization ; Polymorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The non-major histocompatibility complex (MHC)-encoded CD1 family has recently emerged as a new antigen-presenting system that is distinct from either MHC class I or class II molecules. In the present study, we determined the genomic structure of the rat CD1 locus. It was extremely similar to mouse CD1 genes, especially to CD1D1. The 5′ flanking region of the CD1 gene contained the binding motifs for two cytokine-inducible transcription factors, NF-IL2-A and NF-IL6. Some regulatory elements found in MHC class I genes (enhancer A, enhancer B, and the IFN response element) were absent. It is of interest that a tyrosine-based motif for endosomal localization found in the human CD1b cytoplasmic tail was encoded by a single short exon which was conserved in all CD1 molecules except for CD1a. Southern blot and direct sequencing analyses of inbred rat strains suggested very limited polymorphism in the 5′ region where a hydrophobic ligand-binding groove is encoded; a single base substitution resulted in amino acid alteration of alanine (GCT) to valine (GTT) at codon 119. Comparison of the overall exon-intron organization of CD1 genes revealed that the length of the intron was also characteristic to each of the two classes of CD1 genes, classic CD1 and CD1D; such categorization has hitherto been made according to the sequence similarity of the coding region. This finding provides further support for the hypothesis that the two classes have different evolutionary histories. In contrast to the complete absence of the classic CD1 in rats and mice, the entire region of nonpolymorphic CD1D has been conserved through mammalian evolution. Similar functional properties of rodent CD1 and human CD1d are implied.
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  • 15
    ISSN: 1432-1211
    Keywords: Key words Vβ13 ; CD4/CD8 ratio ; Rat ; Tcrb ; Polymorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Three rat BV13S1 alleles (T-cell receptor β-chain variable gene 13) were characterized by new BV13S1-allele specific monoclonal antibodies (18B1 and 17D5) and sequence analysis of expressed and genomic BV13S1. Two alleles were functional and designated BV13S1A1 present in strains LEW, BUF, PVG, and BV13S1A2 present in BN and WF. Their products differed by six amino acids, two of them in complementarity-determing region (CDR)1 and one in CDR2. A third nonfunctional allele, BV13S1A3P, was found in strains F344 and DA. Apart from a single nucleotide insertion, it was identical to BV13S1A2. All 12 rat strains tested showed association of TCRBC1 with BV8S2/4 alleles but not with the BV13S1 alleles, which may reflect a different gene order of the rat BV compared to mouse. BV13S1A1-encoded T-cell receptors (TCRs) which bind both monoclonal antibody (mAb) 18B1 and mAb 17D5 are over-represented in the CD4 lymphocyte subset. BV13S1A2-encoded TCRs which are stained by mAb 18B1 but not by mAb 17D5 show a slight CD8-biased expression. Preferential usage of BV13S1A1-positive TCRs by CD4 but not by CD8 cells in (LEW×WF)F1 hybrids and cosegregation of BV13SA1 and increased frequency of BV13S1 TCR-positive CD4 cells in a (LEW×BN)×BN backcross suggest structural differences of the two allelic products as the reason for their contrasting CD4/CD8 subset bias.
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  • 16
    ISSN: 1432-1211
    Keywords: Key words Eotaxin ; Chemokine ; Eosinophil ; Lung ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 17
    ISSN: 1432-1211
    Keywords: Key words RT1.S3 ; Grc ; MHC ; Class I ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 18
    ISSN: 1432-1211
    Keywords: Key words CD4 ; Rat ; LEC ; thid ; Chromosomal mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 19
    ISSN: 1432-1211
    Keywords: Key words Class II MHC sequence ; Rat ; Cloning ; RT-PCR ; Polymorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 20
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    Biology and fertility of soils 21 (1996), S. 293-302 
    ISSN: 1432-0789
    Keywords: Ammonium excretion ; Azorhizobium caulinodans ; Auxine 2.4-Dichlor-phenoxy-acetic acid ; Nitrogen fixation ; Paranodulation ; Rice ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Rice seedlings developed nodule-like tumors (para-nodules) along primary and secondary roots when treated with the auxin 2,4-dichlor-phenoxy-acetic acid (2,4-D). Histologically, these tumors appeared as cancerous out-grown lateral-root primordes and were thus comparable with stem nodules of the legume Sesbania rostrata. Azorhizobium caulinodans (a diazotroph known as a specific endophyte of Sesbania rostrata) was introduced and became established inside rice para-nodules and in root tissues around tumor bases. The infection with A. caulinodans followed a typical “crack-entry” invasion at places where paranodule tumors had emerged through the root cortex and epidermis. The bacteria settled with high cell densities in intercellular spaces of the induced tumors and between root cortical cells. Infection of plant cells took place both in the epidermis and in cortical tissue. Intracellularly established A. caulinodans was found inside the cytoplasm, surrounded by membrane-like structures. N2 fixation by tumor-inhabiting Azorhizobium sp. was increased at low O2 tensions (1.5–3 kPa) compared with an untreated control. Only a little activity remained at O2 tensions of 5 kPa and above. The present results confirm that root-tumor induction offers a suitable method of establishing diazotrophs endophytically in the roots of gramineous crops.
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  • 21
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    Biology and fertility of soils 21 (1996), S. 293-302 
    ISSN: 1432-0789
    Keywords: Key words Ammonium excretion ; Azorhizobium caulinodans ; Auxine ; 2 ; 4-Dichlor-phenoxy-acetic acid ; Nitrogen fixation ; Paranodulation ; Rice ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Rice seedlings developed nodule-like tumors (para-nodules) along primary and secondary roots when treated with the auxin 2,4-dichlor-phenoxy-acetic acid (2,4-D). Histologically, these tumors appeared as cancerous out-grown lateral-root primordes and were thus comparable with stem nodules of the legume Sesbania rostrata. Azorhizobium caulinodans (a diazotroph known as a specific endophyte of Sesbania rostrata) was introduced and became established inside rice para-nodules and in root tissues around tumor bases. The infection with A. caulinodans followed a typical “crack-entry” invasion at places where para-nodule tumors had emerged through the root cortex and epidermis. The bacteria settled with high cell densities in intercellular spaces of the induced tumors and betwen root cortical cells. Infection of plant cells took place both in the epidermis and in cortical tissue. Intracellularly established A. caulinodans was found inside the cytoplasm, surrounded by membrane-like structures. N2 fixation by tumor-inhabiting Azorhizobium sp. was increased at low O2 tensions (1.5–3 kPa) compared with an untreated control. Only a little activity remained at O2 tensions of 5 kPa and above. The present results confirm that root-tumor induction offers a suitable method of establishing diazotrophs endophytically in the roots of gramineous crops.
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  • 22
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    Calcified tissue international 25 (1978), S. 179-190 
    ISSN: 1432-0827
    Keywords: Decalcification ; Electron microscopy ; Calcified matrices
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The ultrastructure of calcifying cartilage and bone has been examined under the electron microscope after using three different methods of decalcification. The first was carried out before embedding (by soaking specimens in EDTA or formic acid), the second after embedding (by floating ultrathin sections on formic acid), and the third after embedding (by soaking embedded specimens in EDTA or formic acid), and with later re-embedding. The first procedure invariably induces drastic changes in the fine structure of the cells and calcified matrix, probably as a results of the extraction of organic material along with extraction of mineral. The second and third procedures make it possible to preserve ultrastructural details perfectly in both cells and calcified matrix. Of the two, the third procedure is preferable because of its greater simplicity. In areas that are still calcifying, these post-embedding decalcification techniques reveal the presence of crystal-associated, filamentous organic structures which are not recognizable in specimens decalcified before embedding. These structures, which could have a key role in inducing and regulating crystal formation and growth, are less evident in fully calcified areas (but not at their borders). This may partly be due to the loss of glycan components in the matrix during calcification. The most important determinant, however, seems to be the fact that during calcification the components of the matrix, including collagen fibrils, are involved in an aggregation process which reduces the amounts of free chemical groups available for reaction with the stain solution. Because post-embedding decalcification does not disturb this state of aggregation, the stainability of the matrix and the electron microscopic evidence of its components remain very low.
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  • 23
    ISSN: 1432-0827
    Keywords: Cortisol ; Cortical bone ; Bone biomechanics ; Rat ; Femur
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Doses of 8, 16 (low), 32, 48, 64 (medium), and 150 (high) mg/kg/day of cortisol were administered to groups of 8 growing rats each during 16 days, and their femurs were then submitted to 3-point bending tests at low strain rate. Low doses had no effect. Medium doses, previously shown to improve calcium (Ca) balance and weight gain in the species, augmented diaphyseal elastic and ultimate strength, stiffness, and plastic-to-elastic deformation ratio with respect to untreated controls. This effect was achieved either by enhancing bone mass (volume, sectional moment of inertia, wall/lumen ratio) without changes in material quality parameters (32 mg/kg/day) or, conversely, by increasing bone tissue mechanical properties (stress, modulus of elasticity) not affecting bone geometry (48 and 64 mg/kg/day). The highest dose, known to depress Ca balance and weight gain, impaired diaphyseal mechanical performance in controls by substantially reducing bone mass without major variation in bone material properties, that is, developing a true osteopenic state in mechanical terms. The energy elastically absorbed per unit volume (proportional to the risk of comminute fractures) was greater with the highest dose because of enhanced deformability and diminished bone mass. The biphasic dose-response curves obtained, grossly parallel to those previously demonstrated for metabolic actions of cortisol in the same species, showed that biomechanical repercussion of this treatment on bone depends on different, dose-dependent effects which vary independently in temporal course, intensity, and sign.
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  • 24
    ISSN: 1432-0827
    Keywords: Rat ; Osteoblast-like culture ; Ovariectomy ; Estrogens ; Bone blood supply
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Ovariectomy in the rat induces a rapid osteopenia associated with an elevated bone turnover. One hundred and twenty-day-old rats were ovariectomized (OVX) or sham-operated (n=6–8 per group and per time period studied). 45Ca accretion rate and bone blood flow (microspheres trapping technique) in the femurs were determined at 28, 42, 84, and 119 days after ovariectomy. Both parameters were markedly increased by 84 days and subsided thereafter. At the 42nd day, when bone turnover was maximal, bone marrow and trabecular bone cultures were obtained from shamoperated and ovariectomized animals (n=10/group). Proliferation rate of bone marrow cells and trabecular osteoblast-like cells estimated by fibroblast colony-forming units (FCFU) efficiency and cell counting was markedly increased in primary and secondary cultures in ovariectomy. These data fitted well with the enhanced number of osteoblasts observed in situ in the long bone metaphyses of estrogen-depleted animals. As estrogens were shown in the literature to inhibit proliferation of the red cell line and of other hemopoietic lines, it is possible that estrogens, through a general mechanism, inhibit hemopoietic and stromal lines and also the proliferation of bone marrow-derived trabecular bone cells.
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  • 25
    ISSN: 1432-0827
    Keywords: Bone alkaline phosphatase ; Rat ; Vitamin D ; Sorbitol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In the femoral extremities of the adult rat containing the metaphysis, the epiphyseal cartilage, and the epiphysis, four alkaline phosphatase (AP) forms were distinguished on polyacrylamide gel electrophoresis. Two soluble forms were present in the 160,000 g supernatant: one of Mr 165 kDa and another of Mr 110–115 kDa, which exhibited a strong catalytical activity. Moreover, from the pellet, three membrane-bound forms of Mr 130, 110–115, and 100 kDa could be extacted with sodium deoxycholate. When denaturated AP was visualized by postelectrophoretic autoradiography of the phosphorylated intermediates, subunits always appeared as three monomers of Mr 75–80, 60–70, and 50–60 kDa. As four native forms but only three types of subunits were found to be present in the femur, it seems that, apart from homodimers, some heterodimers could also occur. Three types of diets were administered to three groups of rats for 5 weeks. Two are known to disturb bone mineralization: (1) a vitamin D3-deficient diet, and (2) the same as (1) but enriched with 12% sorbitol. The third was a normal diet containing vitamin D3. Concerning the effects on AP of dietary sorbitol and the vitamin D3-deficient diet, it was found that rats receiving the diet supplemented with sorbitol showed a substantial rise in the activity of the Mr 165 kDa form with the concomitant appearance of a new monomer of Mr 100 kDa. In contrast, rats fed the vitamin D3-deficient diet always displayed an increase in enzyme activity, principally of the Mr 100 and 110 kDa forms. In conclusion, the femur extracts of normal rats contained different forms of AP: either soluble 110–115 and 165 kDa forms or membrane-bound 130, 110–115, and 100 kDa forms. The administration of sorbitol-enriched diet induced a marked increase of the 165 kDa form whereas the administration of vitamin D3-deficient diet increased the 100 and 110 kDa forms.
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  • 26
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    Calcified tissue international 50 (1992), S. 524-526 
    ISSN: 1432-0827
    Keywords: Verapamil ; Bone ; Osteopenia ; Rat ; Female ; Intestinal calcium absorption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Verapamil inhibits the intestinal absorption of calcium (Ca) and increases serum parathyroid hormone in rats. The effects of verapamil on bone tissue after long-term treatment is, however, not well described. Adult female and male Sprague-Dawley rats received verapamil in their drinking water at a dosage of 0.075 mg/ml (low dose) or 0.75 mg/ml (high dose) for 12 weeks; control rats received only drinking water. All rats were fed a diet containing 0.1% Ca and 0.5% P. In female rats, the amount of bone ash per volume was significantly reduced from 0.742 g/ml in controls to 0.713 g/ml after low-dose treatment of verapamil, and to 0.667 g/ml following high-dose treatment (P〈0.01). The tibial length was increased from 39.7 mm in controls to 40.3 mm or to 40.7 mm after low or high doses (P〈0.01). The tibial volume increased from 0.385 ml in controls to 0.397 ml after low doses and to 0.429 ml after high doses (P〈0.01). In contrast, in male rats the amount of bone ash per volume was significantly increased from 0.578 g/ml in controls to 0.580 g/ml after low doses and to 0.620 g/ml after high doses of verapamil (P〈0.01). The tibial bone volume in males as decreased from 0.633 ml in controls to 0.641 ml after low doses and to 0.583 ml after high doses (P〈0.05). The tibial length in the males was not changed by verapamil. The intestinal absorption of Ca was reduced in male rats from 5.28 in controls to 4.03 (serosa/mucosa) after low-dose treatment and to 2.46 after high-dose treatment with verapamil (P〈0.05). In female rats, the intestinal absorption of Ca did not change after verapamil treatment. Thus, chronic treatment with verapamil in female rats induced osteopenia whereas in male rats bone growth was inhibited.
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  • 27
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    Calcified tissue international 51 (1992), S. 30-34 
    ISSN: 1432-0827
    Keywords: PTHRP ; Anabolic ; Rat ; Bone ; Potency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Parathyroid hormone-related protein (PTHRP) has recently been purified from human tumors associated with the syndrome of humoral hypercalcemia of malignancy. The gene encoding PTHRP has been cloned, and based on predicted amino acid sequence, polypeptides comprising the first 36 [36Tyr(1–36) PTHRP amide] and 74 [(1–74)PTHRP] amino acids have been synthesized. Human (h) PTHRP (1–36) and (1–74) are potent bone-resorbing agents, and are catabolic for bone in vivo when given continuously at high doses. Bovine parathyroid hormone (bPTH) (1–34) is also catabolic for bone at high dose levels, but when given in low doses for weeks to months, it is anabolic. Although PTHRP possess several PTH-like properties in bone, hPTHRP (1–34) is reported to be only weakly anabolic in vivo. As polypeptide length influences PTHRP action, we evaluated hPTHRP(1–74) as an anabolic agent for bone in vivo. Twenty-four 4-week-old male Sprague-Dawley rats were given daily subcutaneous injections of hPTHRP(1–74) (1 and 2 nmol/100 g body weight, bw), bPTH(1–34) (4 nmol/100 g bw) or vehicle. Rats were sacrificed on day 12, and serum calcium, phosphorus, and 1,25 dihydroxyvitamin D and femoral bone dry weight, calcium content, and hydroxyproline content were measured. Serum calcium and phosphorus were equivalent in all groups. A significant increase in dry bone weight was observed in both PTHRP-treated groups compared with controls. PTHRP also caused a significant, dose-dependent increase in bone calcium and hydroxypro-line content. Results of these studies indicate that PTHRP (1–74) is anabolic for bone in vivo when administered at low-dosage levels for a prolonged period.
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    Calcified tissue international 52 (1993), S. 361-364 
    ISSN: 1432-0827
    Keywords: Calcitonin ; Sustained release ; Copolymer depot ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Studies were carried out to determine whether monolithic depot formulations, prepared using lactide:glycolide copolymers, could be used to administer salmon calcitonin (sCT) to rats in vivo. Formulations containing 2, 5, or 10% (w/w) sCT were administered subcutaneously to female Wistar strain rats. Release of sCT was determined by measurement of peptide in plasma using a specific radioimmunoassay and by measurement of residual sCT in the depots after recovery at postmortem. Plasma calcium concentrations and cumulative weight gain of the animals were used to measure pharmacological effects of the released sCT. Release of sCT from the depots was controlled by the copolymer and was sustained for periods up to 10 days. However, the release of sCT from the depots did not significantly alter plasma calcium concentrations, and effects on cumulative weight gain were small and transient. Peptide loading of the formulations was shown to modify sCT release. Maximal release of sCT from depots containing 10% peptide occurred over a 7 to 14-day period postadministration, with 5% sCT release occurred between days 11 and 14, and with 2% sCT, the period of maximal release was between days 11 and 18. Release of peptide from the depots was essentially complete by 21 days postadministration irrespective of the peptide loading. These data suggest that lactide:glycolide copolymer depots may have application for the convenient clinical administration of sCT in metabolic bone diseases.
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  • 29
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    Calcified tissue international 51 (1992), S. 137-142 
    ISSN: 1432-0827
    Keywords: Swimming exercise ; Bone histomorphometry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary To specify the exercise-induced changes on different skeletal sites, the effect of a 5-week endurance swin training was studied in rats. Eighteen Lyon strain (Sprague-Dawley) 5-week old female rats were divided into nine sedentary and nine swimming rats. Each swim training session was increased by 15 minutes from 2–6 hours per day. A histomorphometric study was performed at the primary and secondary spongiosa of the distal femur and at the secondary spongiosa of lumbar and thoracic vertebral bodies. After training, bone loss was observed in the secondary spongiosa of lumbar vertebral bodies (24.7%) and in the primary spongiosa of distal femur (15.2%). A tendency to bone loss was also detected in the secondary spongiosa of distal femur (10.8%), whereas no change was detected in thoracic vertebral bodies. In secondary spongiosa, bone loss was accompanied with a thinning of trabeculae. Total eroded surfaces and osteoid surfaces were significantly decreased in the three studied skeletal sites, suggesting a decreased bone turnover. The decreased thickness of osteoid seams in both lumbar vertebrae and distal femur could mean that the osteoblastic activity has also been altered at the cell level, leading to thinning of trabeculae. Five-week swim training with such duration and intensity of exercise appears unable to increase bone volume in rats and, therefore, causes adverse effects. The three studied bones seemed to adapt differently to experimental conditions. The lack of ground reaction forces induced by water immersion might have contributed to the observed bone loss. “Normal” gravity would be an important cofactor in the osteogenic effects of exercise.
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  • 30
    ISSN: 1432-0827
    Keywords: Decalcification ; Electron microscopy ; Bone matrix ; Bone glycoproteins
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    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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  • 31
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    Archives of microbiology 107 (1976), S. 99-107 
    ISSN: 1432-072X
    Keywords: Piptocephalis unispora ; Mucorales ; Kickxellaceae ; Electron microscopy ; Germination ; Spore swelling ; Sporangiospore
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    Topics: Biology
    Notes: Abstract Germination of the sporangiospore of Piptocephalis unispora Benjamin, observed by means of light and electron microscopy, involved the formation of a new inner wall which became continous with the inner layer of the wall of the germ tube. The outer wall layer of the germ tube was continous with the original inner wall layer of the dormant spore. Preliminary details of appressorium structure were noted. Nutritional experiments indicated that sporangiospores required external sources of utilisable nitrogen and carbon compounds for maximal swelling and germ tube production. Limited development occurred when either nutrient was supplied singly. Comparison of germination of the asexual spore with that in other Mucorales, especially the Kickxellaceae, has been made, and the merosporangial status in P. unispora discussed.
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  • 32
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    Archives of microbiology 107 (1976), S. 113-114 
    ISSN: 1432-072X
    Keywords: Achlya ; Electron microscopy ; Nuclear microfilaments ; Antheridia ; Mycology
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    Topics: Biology
    Notes: Abstract This is the first report of intranuclear microfilaments within gametangial nuclei of oömycetous fungi. Longitudinal sections of four to six microfilaments were frequently observed in meiotic antheridial nuclei of Achlya ambisexualis. Each microfilament measured approximately 7–10 nm in diameter. Spindle tubules (25 nm in diameter) were also observed within some of the nuclei possessing microfilaments.
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  • 33
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    Archives of microbiology 107 (1976), S. 167-182 
    ISSN: 1432-072X
    Keywords: Ectothiorhodospira mobilis ; Photosynthetic membranes ; Electron microscopy ; Isolation of membranes ; Structure of membranes ; Composition of membranes
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    Topics: Biology
    Notes: Abstract The lamellar membrane stacks of Ectothiorhodospira mobilis were isolated and purified by a combination of lysozyme and osmotic shock treatment, followed by differential and density gradient centrifugation. Preparations of lamellar membranes were enriched at least 2.4-fold in the ratio of bacteriochlorophyll a to protein. Thin-sectioning, negative staining, platinumcarbon shadowing and freeze-etching were used to study the architecture of the membrane units. Both platinum-carbon shadowing and freeze-etching showed the outer surfaces of the isolated lamellar membrane stacks to be relatively smooth. Particles averaging 7 nm in diameter were seen on several faces following freeze-ctching. Non-polar amino acids amounted to 60% of the total amino acid composition. Lipids constituted 32% of the membrane dry weight. Phosphatidyl ethanolamine and diphosphatidyl glycerol were the major phospholipids. Fatty acids of 10–15 carbons represented a small fraction of both membrane and whole cell fatty acids. Monoenes constituted 36% of the total membrane fatty acids and 38.4% of the total whole cell fatty acids. The major fatty acids of both whole cells and purified membranes were C16:0, C18:1 and cyclopropane C19:0.
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  • 34
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    Archives of microbiology 107 (1976), S. 313-320 
    ISSN: 1432-072X
    Keywords: Micrococcus radiophilus ; Micrococcus radioproteolyticus ; Bacterial cell walls ; Fine structure ; Electron microscopy ; Taxonomy
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    Topics: Biology
    Notes: Abstract The radiation resistant bacteria Micrococcus radiophilus and M. radioproteolyticus were studied by thin sectioning and freeze-etching techniques and the two species were found to be similar in the fine structure. The only significant difference was in the appearance of the surfaces of the cell walls in freeze-etched preparations. Since the two species, together with M. radiodurans, possess a unique cell wall structure and a cell wall peptidoglycan, which is different from that of other micrococci and Gram-positive cocci, it is recommended that they be reclassified into a new genus.
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  • 35
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    Archives of microbiology 108 (1976), S. 55-64 
    ISSN: 1432-072X
    Keywords: Bdellovibrio ; Spirillum ; Cell wall ; Bdelloplast ; Lipoprotein ; Peptidoglycan ; Electron microscopy
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    Topics: Biology
    Notes: Abstract In both freeze-etched and critical-point dried preparations examined by transmission and scanning electron microscopy, respectively, the outer surfaces of the cells of Spirillum serpens VHL assume a wrinkled appearance 10–15 min after challenge by Bdellovibrion bacteriovorus 109D. This wrinkling effect is believed (on circumstantial evidence) to be caused by the bdellovibrio's disruption of the cell wall lipoprotein of the Spirillum. With the exception of those topological changes caused by wrinkling, the outer membrane of the Spirillum cell wall retains a normal appearance as viewed in freeze-etched preparations, even after the Spirillum cell has been converted into a bdelloplast. Although the peptidoglycan layer of the Spirillum cell presumably is weakened somewhat by the invading Bdellovibrio, evidence obtained from freeze-fractured preparations of Spirillum bdelloplasts suggests that the peptidoglycan remains as a discrete cell wall layer, even though the Spirillum cell wall apparently has lost much of its rigidity. That the peptidoglycan backbone remains essentially intact, even after the Spirillum cell has been entered by the Bdellovibrio, is supported by the observation that the soluble amino sugar content of the culture medium, as determined by chemical analysis, does not rise even 5.0 h after the association of the Bdellovibrio with the Spirillum has begun.
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  • 36
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    Archives of microbiology 116 (1978), S. 133-139 
    ISSN: 1432-072X
    Keywords: Lagenisma ; Coscinodiscus ; Infection ; Endosymbiotic bacteria ; Tip growth ; Wall-less thallus ; Host-parasite interface ; Membranes ; Electron microscopy
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    Topics: Biology
    Notes: Abstract Lagenisma coscinodisci is diplanetic and has two different cyst stages. The secondary cyst has a uniform cell wall of fibrillar material. It attaches to a Coscinodiscus frustule and germinates with an infection tube. The cyst becomes filled with an enlarging expulsion vacuole. The infection tube penetrates the diatom cell between the cingula. Inside the host cell the fungus grows as an irregularly branched wall-less thallus. In the hyphae apical vesicles are lacking. The infection tube is plugged by wall material. There are no microtubules which might participate in the morphogenesis of the thallus. The plasmalemma of the diatom is pushed inward but not pierced by the fungus. Along the host-parasite interface it lies closely paralled to the Lagenisma plasmalemma which is extremely straight here and measures about 10 nm instead of about 5–6 nm at the surfaces of other stages. The Coscinodiscus plasmalemma disintegrates at about the same time when the cytoplasm breaks down. The fungus allows bacteria to enter the diatom; there are also endosymbiontic bacteria in unattacked cells — The growth mechanisms are discussed and the host-parasite interface is compared with that of other fungi.
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  • 37
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    Archives of microbiology 121 (1979), S. 9-15 
    ISSN: 1432-072X
    Keywords: R-Bodies ; Kappa particles ; Free-living hydrogen bacteria ; Induction ; Electron microscopy ; Chemical composition ; Defective prophages ; Plasmids
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    Topics: Biology
    Notes: Abstract R-Bodies have been found in a recently isolated pseudomonas-like free-living hydrogen oxidizing bacterium. Their isolation, fine structure and chemical composition are described and compared with the R-bodies from the kappa particles (Caedobacter), obligate endosymbionts of Paramecium aurelia. The 2K 1 R-bodies exhibited essential characteristics of the kappa R-bodies; however, their size and some other structural aspects proved that they represent a new type of R-bodies. The presence of phage tail-like particles in cells induced with Mitomycin C is in favour of the hypothesis that the R-bodies might be coded by defective prophages, or by extrachromosomal elements.
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  • 38
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    Keywords: Acetobacter suboxydans ; Bacteriophage A-1 ; Restriction ; Modification ; Electron microscopy
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    Notes: Abstract A bacteriophage ofAcetobacter suboxydans was isolated and found to correspond to type A phage according to Bradley's classification. The phage contains double stranded DNA. The length of the latency period and burst size could not be precisely determined because of apparent non-synchronous release of phage from single infective cycles. The host range was determined using 24 strains ofAcetobacter andGluconobacter species. Evidence for a probable occurence of host determined restriction and modification was obtained withAcetobacter suboxydans strain ATCC 621. The phage is designated A-1 and it is the first one to be reported forAcetobacter.
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  • 39
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    Archives of microbiology 123 (1979), S. 173-181 
    ISSN: 1432-072X
    Keywords: Bacillus subtilis ; Cell cycle ; DNA replication ; Cell division ; Electron microscopy
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    Topics: Biology
    Notes: Abstract Bacillus subtilis strain Marburg was grown exponentially with a doubling time of 65 min. To follow the time course of various cell cycle events, cells were collected by agar filtration and were then classified according to length. The DNA replication cycle was determined by a quantitative analysis of radioautograms of tritiated thymidine pulse labeled cells. The DNA replication period was found to be 45 min. This period is preceded and followed by periods without DNA synthesis of about 10 min. The morphology and segregation of nucleoplasmic bodies was studied in thin sections. B. subtilis contains two sets of genomes. DNA replication and DNA segregation seem to go hand in hand and DNA segregation is completed shortly after termination of DNA replication. Cell division and cell separation were investigated in whole mount preparations (agar filtration) and in thin sections. Cell division starts about 20 min after cell birth; cell separation starts at about 45 min and before completion of the septum.
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  • 40
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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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  • 41
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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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    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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  • 42
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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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    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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  • 43
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    Keywords: Bacteriolysis ; Penicillin ; Autolysis ; Cell wall ; Electron microscopy ; Staphylococcus aureus
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    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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  • 44
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    Archives of microbiology 163 (1995), S. 300-306 
    ISSN: 1432-072X
    Keywords: Nitrosomonas europea ; Hydroxylamine oxidoreductase (HAO) ; Electron microscopy ; Electron spectroscopic imaging ; Quaternary structure
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    Notes: Abstract The hydroxylamine oxidoreductase from Nitrosomonas europaea was prepared to apparent electrophoretic homogeneity. Electron microscopy of negatively stained preparations of the sample revealed an overall diameter of about 8.8 nm of the enzyme particle. The native structure was determined as a tetrahedron-like assembly of identical subunits exhibiting four protein masses.
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  • 45
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    Archives of microbiology 105 (1975), S. 193-199 
    ISSN: 1432-072X
    Keywords: Bean ; Rust ; Haustorium ; Sheath ; Autoradiography ; Infection ; Electron microscopy ; Phaseolus vulgaris ; Uromyces phaseoli
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    Notes: Abstract Tritium labeled uredospores of Uromyces phaseoli were produced be feeding the host, Phaseolus vulgaris, with 3H-orotic acid. These spores were allowed to germinate on and to penetrate into a bean leaf. 24 hrs after inoculation, the bean rust had formed the first haustorium. All fungal structures, including the fungus walls, were heavily labeled. No label could be detected in the cells that had come into contact with the hyphae. In the infected host cell, the haustorium was labeled heavily, but the sheath around the haustorium and the host cell remained free of label. These results indicate that no detectable amounts of label leach from the bean rust into the host at this stage of infection although it is known that the rust takes up many metabolites. Since the sheath remains free of label and all fungal structures are evenly labeled, it is concluded that the sheath is formed by the host.
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  • 46
    ISSN: 1432-072X
    Keywords: Acidaminococcus fermentans ; Glutamate fermentation ; Electron microscopy ; Immunocytochemistry ; Post-embedding labelling ; Antibody-gold complexes ; Protein A-gold complexes
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    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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  • 47
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    Keywords: Aggregation substance ; Enterococcus faecalis ; Electron microscopy ; Field emission scanning electron microscopy ; Immunogold labelling technique ; Sex pheromone system
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    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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  • 48
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    Keywords: Soluble NAD-dependent hydrogenase ; Alcaligenes eutrophus ; Nocardia opaca ; Electron microscopy
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    Notes: Abstract The soluble NAD-dependent hydrogenase (hydrogen-NAD oxidoreductase, EC 1.12.1.2), consisting of four non-identical subunits, was isolated from Alcaligenes eutrophus H16 and from Nocardia opaca 1b and analyzed by a HPLC gel permeation technique and electron microscopy. The tetrameric enzyme particles from both origins, as determined from negatively stained electron microscopic samples, were found to be elongated and very similar in shape and size. The A. eutrophus enzyme was measured in more detail. It exhibited dimensions of 12.7 nm by 5.5 nm (axial ratio 2.3:1). Dissociation into smaller particles and unspecific aggregation combined with partial inactivation were observed in the presence of the inhibitor NADH. Kept in buffer without added nickel, the enzyme was partially dissociated. Reassociation of tetramers without restored enzyme activity was achieved by addition of 0.5 mM NiCl2. A working model for the structural organization of the tetrameric enzyme particle is presented.
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  • 49
    ISSN: 1432-072X
    Keywords: Lactobacillus ; Medium composition ; Metal cations ; Electron microscopy ; Protoplast-like forms
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    Notes: Abstract The growth of some locally isolated Lactobacillus strains forming D(-) or L(+) lactic acid, Lactobacillus helveticus ATCC 15009 and Lactobacillus delbrueckii subsp. bulgaricus ATCC 11842 was examined in different media. L. helveticus and Lactobacillus LBL strains formed atypical protoplast-like cells in LAPT medium, sensitive to SDS and proteinase. Specific morphological changes in the cell wall structure of these variants were revealed by transmission and scanning electron microscopy. The effect of glucose and various salts on their appearance was investigated. The prevalent role of metal cations, especially of Mg2+, was established.
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  • 50
    ISSN: 1432-072X
    Keywords: d-Ribulose 1,5-diphosphate carboxylase ; Oxygenase activity ; Quaternary structure ; Electron microscopy ; Alcaligenes eutrophus
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    Notes: Abstract d-Ribulose 1,5-diphosphate carboxylase has been purified from autotrophically grown cells of the facultative chemolithotrophic hydrogen bacteriumAlcaligenes eutrophus. The enzyme was homogeneous by the criteria of polyacrylamide gel electrophoresis. The molecular weight of the enzyme was 505000 determined by gel filtration and sucrose density gradient centrifugation, and a sedimentation coefficient of 18.2 S was obtained. It was demonstrated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis that the enzyme consists of two types of subunits of molecular weight 52000 and 13000. Electron microscopy on the intact and the partially dissociated enzyme lead to the construction of a model for the quaternary structure of the enzyme which is composed of 8 large and 8 small subunits. The most probable symmetry of the enzyme molecule is 4:2:2. Michaelis constant (K m ) values for ribulose 1,5-diphosphate, Mg2-, and CO2 were 0.59 mM, 0.33 mM, and 0.066 mM measured under air. Oxygen was a competitive inhibitor with respect to CO2 suggesting that the enzyme also exhibits an oxygenase activity. The oxygenolytic cleavage of ribulose 1,5-diphosphate was shown and a 1:1 stoichiometry between oxygen consumption and 3-phosphoglycerate formation observed.
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  • 51
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    Archives of microbiology 108 (1976), S. 231-242 
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    Keywords: Phytophthora ; Penetration ; Eucalypts ; Roots ; Electron microscopy ; Appressoria ; Plugs
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    Topics: Biology
    Notes: Abstract The mechanisms of penetration of Phytophthora cinnamomi Rands into seedling eucalypt roots were studied by light and electron microscopy. Culture grown seedlings of root-rot tolerant Eucalyptus st johnii and root-rot susceptible Eucalyptus obliqua were inoculated with both zoospores and mycelium. Zoospores encysted on roots of both species and the germ tubes penetrated without the formation of appressoria. Swellings, previously described as appressoria, were formed when the germ tube was slow to enter the host by intracellular penetration. Vegetative hyphae penetrated both inter- and intracellularly into the zones of root elongation and differentiation, often through root hairs. Evidence of hydrolysis of the host cell-wall at the point of penetration was observed in electron micrographs. Several hours after the germ tube penetrated the epidermis, a thick plug of amorphous material formed in the germ tube slightly below the level of the outer walls of the epidermal cells, sealing off the hypha within the root. Behaviour of zoospores and germ tubes and the mechanism of penetration were similar on both hosts. Micrographs do not suggest any kind of a hypersensitive reaction by the host cells during the early stages of infection.
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  • 52
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    Archives of microbiology 115 (1977), S. 185-198 
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    Keywords: Synechococcus lividus ; Cyanobacteria ; Carbon dioxide ; Electron microscopy ; Bleaching-regreening
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    Topics: Biology
    Notes: Abstract The effect of carbon dioxide on pigment and membrane content in Synechococcus lividus was studied by depriving cells of CO2 and examining cell populations biochemically and by electron microscopy. After 120 h of CO2 deprivation, S. lividus lost all detectable chlorophyll a and C-phycocyanin. Such bleached cultures were “mustard yellow”, the result of approximately 1.8 times more carotenoid per cell than green control cultures. Although cells from beached cultures appeared morphologically identical to control green cells when examined by light microscopy, electron microscopic examination revealed them to be devoid of detectable thylakoid membrane. Thylakoid membrane could not be recovered by physical isolation or revealed by freeze etching of bleached S. lividus. In addition, inclusion bodies characteristically found in S. lividus were also absent. Reintroduction of CO2 into bleached cultures resulted in a rapid resynthesis of both chlorophyll a and C-phycocyanin. Electron microscopic examination of these regreening cultures revealed that thylakoid membrane was also rapidly resynthesized. Growth of regreened cultures did not occur until there was the synthesis of a full complement of chlorophyll a, C-phycocyanin, and thylakoid membrane. A time course study of the cytological events occurring during bleaching and regreening is presented.
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  • 53
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    Archives of microbiology 109 (1976), S. 21-30 
    ISSN: 1432-072X
    Keywords: Electron microscopy ; Allomyces ; Gametes ; Fertilization ; Membrane fusion
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    Topics: Biology
    Notes: Abstract The gametes and the process of fertilization were examined by light and electron microscopy in the lower eukaryote Allomyces macrogynus. Differences in gamete morphology included the overall larger size and the presence of a larger nuclear apparatus, along with the association of a side-body complex and many more mitochondria in the female gamete. In this species of Allomyces, fertilization was initiated by contact and fusion of specialized regions of the gamete plasma membranes resulting in a binucleate fusion cell surrounded by plasma membrane contributed by both partners. Following plasmogamy, nuclear fusion was initiated by multiple nuclear membrane contacts between adjacent outer membranes. Following inner membrane fusion, small nucleoplasmic bridges were observed which presumably fused with one another and resulted in a single bridge which widened, forming the mature diploid nucleus. After karyogamy, fusion of the nuclear caps did not always occur and zygotes with and without fused caps were observed. Coalescence of the nucleoli completed the events of fertilization, forming a zygote with a single nuclear apparatus (sometimes with two caps) and two flagella. These observations are discussed in relation to fertilization mechanisms and compared to fertilization in other organisms.
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  • 54
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    Archives of microbiology 118 (1978), S. 67-69 
    ISSN: 1432-072X
    Keywords: Corynebacterium autotrophicum ; Outer Membrane ; Lipopolysaccharides ; Biochemical analysis ; Polymyxin B ; Electron microscopy
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    Notes: Abstract Lipopolysaccharides (LPS) from Corynebacterium autotrophicum were isolated and analyzed. Autotrophically grown cells contained 2–5 mg of partly purified LPS per g dry weight of lyophilized cells. Serological cross reaction with Lipid A antigen of Salmonella minnesota confirmed the presence of LPS in C. autotrophicum. Electron microscopy of negatively stained Polymyxin B-treated cells showed formation of blebs on the Outer Membrane indicating an interaction of Polymyxin B specifically with LPS. Up to now, no Gram-positive organisms are known which contain any LPS. Thus, C. autotrophicum, though giving opposite results when the Gram-staining reaction was applied by several authors, has to be classified into the group of Gram-negative bacteria.
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  • 55
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    Archives of microbiology 119 (1978), S. 303-304 
    ISSN: 1432-072X
    Keywords: Vibrio cholera phage group II ; Properties ; Electron microscopy
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    Topics: Biology
    Notes: Abstract The basic physical, chemical and physiological properties of a group II cholera phage belonging to Mukerjee's classification has been described.
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  • 56
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    Archives of microbiology 118 (1978), S. 305-308 
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    Keywords: Carotenoid mutant strain R-26-Rhodopseudomonas sphaeroides ; Electron microscopy ; Intracellular membranes
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    Notes: Abstract Stained thin-sections and freeze-fractured preparations of the carotenoid-less mutant strain R-26 of Rhodopseudomonas sphaeroides grown photosynthetically revealed 2 morphological kinds of intracellular membrane systems- spherical vesicles distributed throughout the cytoplasm and lamellae confined to the periphery of the cell. The lamellar membranes appeared to be large, flattened vesicles.
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  • 57
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    Keywords: Alcohol dehydrogenase ; Acetaldehyde dehydrogenase ; Clostridium kluyveri ; Electron microscopy ; Polygonal bodies ; Enzyme complex
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    Notes: Abstract The alcohol-acetaldehyde dehydrogenase complex of Clostridium kluyveri has been separated from contaminating β-hydroxybutyryl-CoA dehydrogenase by repeated precipitation with manganese and ammonium sulfate. Mn++ was required for maximum alcohol dehydrogenase activity. The molecular weight of the enzyme complex was 194,000 as determined by sucrose density gradient centrifugation. The enzyme complex has been shown to contain two types of subunits with molecular weights of 55,000±2,600 and 42,000±1,200, respectively which are arranged in “H”-shaped particles. In solutions with an ionic strength above 25 mM the enzyme complex precipitated in the form of lumps as has been shown with specific ferritin-conjugated antibodies. These lumps are assumed to be aggregated polygonal bodies present in C. kluyveri.
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  • 58
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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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    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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  • 59
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    Keywords: Allomyces ; Zoospores ; Cell wall ; Chitin ; Gamma particle ; Encystment ; Electron microscopy ; Calcofluor
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    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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  • 60
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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
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    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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  • 61
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    Archives of microbiology 130 (1981), S. 125-128 
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    Keywords: Rhodospirillum rubrum ; Chromatophores ; Reaction centers ; Liposomes ; Electron microscopy
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    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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  • 62
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    Archives of microbiology 142 (1985), S. 333-339 
    ISSN: 1432-072X
    Keywords: Photosynthesis ; Membrane structure ; Electron microscopy ; Photosynthetic bacteria
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    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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  • 63
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    Archives of microbiology 157 (1992), S. 381-388 
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    Keywords: PhiX174 ; Bacterial lysis ; Escherichia coli ; Electron microscopy ; Membranes ; Cell envelope
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    Notes: Abstract Expression of cloned gene E of bacteriophage PhiX174 induces lysis by formation of a transmembrane tunnel structure in the cell envelope of Escherichia coli. Ultrastructural studies of the location of the lysis tunnel indicate that it is preferentially located at the septum or at polar regions of the cell. Furthermore, the diameter and shape of individual tunnel structures vary greatly indicating that its structure is not rigid. Apparently, the contours of individual lysis tunnels are determined by enlarged meshes in the peptidoglycan net and the force produced at its orifice, by the outflow of cytoplasmic content. Once the tunnel is formed the driving force for the lysis process is the osmotic pressure difference between cytoplasm and medium. During the lysis process areas of the cytoplasmic membrane which are not tightly attached to the envelope are extended inward by the negative pressure produced during lysis. After cell lysis external medium can diffuse through the lysis tunnel filling the inner cell space of the still rigid bacterial ghosts.
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  • 64
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    Archives of microbiology 159 (1993), S. 114-118 
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    Keywords: Bacillus pulvifaciens ; Vegetative cells ; Spotes ; Ultrastructure ; Electron microscopy
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    Notes: Abstract The ultrastructure of vegetative cells and spores of Bacillus pulvifaciens was studied by CTEM and SEM methods. The vegetative cells are rods, 1.6–4.5 μm long and 0.4–0.6 μm wide, exhibiting typical ultrastructural features of Gram-positive bacteria. The spores are of ellipsoidal shape, 0.6×1.2 μm in size, with six longitudinal ribs reaching up to 130 nm in height. There are satelite ribs on both sides of the longitudinal ribs, reaching up to 20 nm in height. Between the longitudinal ribs, additional transversal ribs were observed in SEM. A special tubular layer, separating the outer and inner coat of the spores, was revealed in ultrathin sections. This layer seems to be a typical ultrastructural feature of Bacillus pulvifaciens spores.
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  • 65
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    Archives of microbiology 162 (1994), S. 267-271 
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    Keywords: Key words     Extremely thermophilic eubacterium ; Calderobacterium hydrogenophilium ; Ultrastructure ; Electron microscopy
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    Notes: Abstract       Calderobacterium hydrogenophilum is an extreme thermophilic, obligately chemoautotrophic, hydrogen-oxidizing bacterium. The cells were shown to be non-motile straight rods of average size 0.4 × 2.5 μm. After negative-staining of the whole cells, no flagella were observed. The multilayered cell wall was of type 1 and possessed a crystalline proteinaceous surface layer exhibiting p4 symmetry. The square unit cells had a lattice constant of approximately 11 nm. Cell division occurred by a constriction mechanism. C. hydrogenophilum differred from a similar hydrogen-oxidizing eubacterium, Hydrogenobacter thermophilus, by the absence of intracytoplasmic membrane structures in chemically fixed cells. However, an electron-dense intracytoplasmic hemispherical structure adhering to the inner membrane was frequently observed.
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  • 66
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    Archives of microbiology 160 (1993), S. 284-287 
    ISSN: 1432-072X
    Keywords: Bacterial glucoamylase ; Clostridium thermosacharolyticum ; Cellular location ; Activity states ; Macromolecular organization ; Electron microscopy
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    Notes: Abstract By application of immunocytochemical techniques at the electron microscope level, glucoamylase was localized to the cell periphery in Clostridium thermosaccharolyticum during and following growth on starch, sucrose or glucose. Levels of immunolabelling were found to be relatively independent of growth substrate and of phase of growth, whereas previous studies had demonstrated strong dependence of glucoamylase activity on growth conditions; previously high levels of glucoamylase activity had been detected after growth on starch (i.e. during the stationary phase after growth) and only very low activities detected during exponential growth and following growth on glucose. The results presented demonstrate that levels of the glucoamylase protein are independent of measurable enzyme activity, and imply that the protein is constitutive. This indicates that the protein can exist in active and inactive states in the cell. By analogy with similar systems, we consider it likely that “maturation” or “activation” of newly synthesized glucoamylase occurs during (or following) transport through the cytoplasmic membrane. Electron microscopy of individual protein molecules which had been subjected to negative staining revealed that the enzyme consists of two domains of approximately equal size which are linked by a “hinge” region.
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    Archives of microbiology 162 (1994), S. 267-271 
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    Keywords: Extremely thermophilic eubacterium ; Calderobacterium hydrogenophilium ; Ultrastructure ; Electron microscopy
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    Notes: Abstract Calderobacterium hydrogenophilum is an extreme thermophilic, obligately chemoautotrophic, hydrogen-oxidizing bacterium. The cells were shown to be nonmotile straight rods of average size 0.4x2.5 μm. After negative-staining of the whole cells, no flagella were observed. The multilayered cell wall was of type 1 and possessed a crystalline proteinaceous surface layer exhibiting p4 symmetry. The square unit cells had a lattice constant of approximately 11 nm. Cell division occurred by a constriction mechanism. C. hydrogenophilum differred from a similar hydrogen-oxidizing eubacterium, Hydrogenobacter thermophilus, by the absence of intracytoplasmic membrane structures in chemically fixed cells. However, an electron-dense intracytoplasmic hemispherical structure adhering to the inner membrane was frequently observed.
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  • 68
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    Keywords: Key words Magnetic bacteria ; Biomineralization ; Magnetite ; 16S rRNA ; In situ hybridization ; Ultrastructure ; Electron microscopy
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    Notes: Abstract Natural enrichments of magnetic bacteria from the Itaipu lagoon near Rio de Janeiro were dominated by coccoid-to-ovoid morphotypes that produced unusually large magnetosomes. To determine the phylogenetic position of these unusual microorganisms, 16S rRNA genes were retrieved from bacteria magnetically separated from sediment of the Itaipu lagoon by in vitro amplification and cloning of PCR products into a plasmid vector. Partial sequencing of the obtained clones revealed two clusters of closely related sequences affiliated to a distinct lineage consisting exclusively of magnetic bacteria within the α-subclass of Proteobacteria. For a detailed phylogenetic analysis, several almost complete sequences of the 16S rRNA genes were determined. One representative clone of each cluster provided a PCR template for the in vitro transcription of group-specific polynucleotide probes complementary to a variable region of the 16S rRNA molecule. At least three different morphotypes of magnetic bacteria were reliably identified by post-embedding hybridization of ultra-thin sections. Electron microscopic analyses of hybridized cells enabled for the first time a detailed description of the morphological variety and ultrastructure of phylogenetically identified, uncultured magnetic bacteria. Two distinct coccoid bacteria were identified by the transcript probe complementary to the 16S rRNA sequence mabrj12, whereas the probe complementary to the sequence mabrj58 allowed the identification of an ovoid morphotype that displayed magnetosomes with the largest volumes observed to date.
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    Archives of microbiology 111 (1976), S. 175-183 
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    Keywords: Serratia marcescens ; (Phage tail) bacteriocin ; Electron microscopy
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    Topics: Biology
    Description / Table of Contents: Zusammenfassung Die Genese eines phagenschwanzähnlichen Bacteriocins in Zellen des Gruppe A-bacteriocinogenen (bA+) Serratia marcescens-Stammer Nr. 16 wurde nach Mitomycin C (MC) Induktion elektronenoptisch untersucht. Dieses Bacteriocin (Gesamtlänge 117 nm) besteht aus einem hohlen Stift mit kontraktiler Scheide. Nach 60 min Induktion wurden in Dünnschnitten stäbchenförmige Bacteriocine identifiziert. Sie erscheinen in drei Aggregationsformen: 1. als hexagonale Einschlüsse, 2. als Bänder dicht nebeneinanderliegender Bacteriocine und 3. als Stapel von übereinanderliegenden Bacteriocinschichten, wenn nach 120 min Induktion ein Maximum von ca. 450 Bacteriocinen pro Zelle erreicht wird. Bacteriocine konnten nach der gleichen Induktionszeit von 60 min auch mit der “in situ lysis technique” nachgewiesen werden. Neben Bacteriocinen traten relativ selten und unregelmäßig auch Phagenköpfe auf. Die Stäbchenform teilungsfähiger Zellen blieb bis zum Auftreten von intracellulären Bacteriocinen erhalten. Ihre Umwandlung in geblähte, sphäroplastenähnliche Zellformen, die nach 120 min Induktion lysierten, war zeitlich korreliert mit Feinstrukturveränderungen der Zellwand.
    Notes: Abstract The biosynthesis of a phage tail-like Bacteriocin by cells of the group A-bacteriocinogenic (bA+ Serratia marcescens strain no. 16 after induction with mitomycin C (MC) was examined electronmicroscopically. This bacteriocin (total length 117 nm) consists of a hollow core and a contractile sheath. At 60 min following induction, rod-like bacteriocin-partieles were identifiable in ultrathin sections. The particles were found to comprise three morphologically different forms of aggregation: 1. hexagonal inclusions, 2. contiguous, bank-like particles, and 3. staples of superimposed layers of bacteriocin particles. At 120 min after induction bA+ cells revealed maximally 450 bacteriocin particles. Similarly, the phage tail particles could be demonstrated with the “in situ lysis technique” at 60 min following induction. Occasionally, phage heads were demonstrable, but in no instance were complete phage particles discernible. Dividing cells of the bA+ strain of S. marcescens maintained their rod-form following induction with MC until intracellular phage tail bacteriocin particles were seen. However, at 120 min after induction, the swollen, sphaeroplast-like cells lysed, an event that could be correlated with fine structural alterations of the cell wall.
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    Archives of microbiology 112 (1977), S. 207-218 
    ISSN: 1432-072X
    Keywords: Cryptophyceae ; Algae ; Hemiselmis rufescens ; Chroomonas ; Cryptomonas ; Mitochondrial complex ; Cristae ; Electron microscopy
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    Notes: Abstract The unitary nature of the mitochondrion and the characteristic flattened finger-like morphology of the cristae were demonstrated in the Cryptophyceae. Hemiselmis rufescens contained an unbranched vermi-form mitochondrion in contrast to the variously branched complex, comprising an interconnected peripheral and central reticulum, in Chroomonas sp. and strains of Cryptomonas. The systematic value of the shape and distribution of the mitochondria in the examined genera was suggested.
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    Archives of microbiology 113 (1977), S. 197-204 
    ISSN: 1432-072X
    Keywords: Gliding bacterium ; Simonsiella ; Oral cavity ; Electron microscopy ; Morphology ; Dorsal-ventral differentiation ; Ultrastructure
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    Notes: Abstract The morphology and ultrastructure of the aerobic, Gram-negative multicellular-filamentous bacteria of the genus Simonsiella were investigated by scanning and transmission electron microscopy. The flat, ribbon-shaped, multicellular filaments show dorsal-ventral differentiation with respect to their orientations to solid substrata. The dorsal surface, orientated away from the substrate, is convex and possesses an unstructured capsule. The ventral surface, on which the organisms adhere and glide, is concave and has an extracellular layer with fibrils extending at right angles from the cell wall. The cytoplasm in the ventral region contains a proliferation of intracytoplasmic membranes and few ribosomes in comparison to the cytoplasm in other parts of the cell. Centripetal cell wall formation is asymmetrical and commences preferentially in the ventral region. Quantitative differences in morphology and cytology exist among selected Simonsiella strains. Functional aspects of this dorsalventral differentiation are discussed with respect to the colonization and adherence of Simonsiella to mucosal squamous epithelial cells in its ecological habitat, the oral cavities of warm-blooded vertebrates.
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    Archives of microbiology 109 (1976), S. 195-197 
    ISSN: 1432-072X
    Keywords: Cell wall ; Peptidoglycan ; Electron microscopy ; Bacillus subtilis
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    Notes: Abstract Isolated cell walls of Bacillus subtilis have a striated appearance in the electron microscope. The structure persists when teichoic acids are removed. It is inferred that the structure bears on the arrangement of the peptidoglycan chains.
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    Archives of microbiology 119 (1978), S. 227-229 
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    Keywords: Cell wall ; Electron microscopy ; Methylomonas albus ; Goblet sub-units
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    Notes: Abstract In surface view, the cell wall complex ofMethylomonas albus possesses a hexagonal pattern of ridges. Thin sections reveal a continuous layer of goblet-shaped elements attached to the outermost surface of the lipopolysaccharide membrane. A possible interpretation of the cell wall complex ofM. albus, based on the fine-structural data is presented.
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  • 74
    ISSN: 1432-072X
    Keywords: Defective lysogeny ; Alcaligenes eutrophus ; Simultaneous isolation technique ; Temperate bacteriophages ; Pseudomonas pseudoflava ; Biological characterization ; Electron microscopy
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    Notes: Abstract Widespread defective lysogeny was detected in Alcaligenes eutrophus by electron microscopic analysis of cultures. Mitomycin C treatment of the cultures resulted in the production of defective (inco-) particles. Polysheaths were produced both with and without induction. With the simultaneous isolation technique six phages were isolated for hydrogen-oxidizing strains of the new species Pseudomonas pseudoflava. The phages were able to replicate under autotrophic conditions and were found to have a very restricted host range. Electron microscopic analysis allowed classification into two structural groups. Group I contained phages with contractile tails; group II contained phages with flexible, noncontractile tails. All but one (gb) of the new phages were shown to be temperate by isolation of lysogens and induction with mitomycin C.
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  • 75
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    Archives of microbiology 123 (1979), S. 101-103 
    ISSN: 1432-072X
    Keywords: Bdellovibrio ; Cyanobacteria ; Marine sponges ; Symbiosis ; Infection ; Electron microscopy
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    Notes: Abstract A bdellovibrio-like bacterium was observed infecting unicellular symbiotic cyanobacteria in two coral reef sponges, Neofibularia irata and Jaspis stellifera. The infecting bacterium, which was located between the cell wall and the cytoplasmic membrane of the cyanobacteria, was similar in size and appearance to previously described bdellovibrios. This observation is believed to extend the host range of the bdellovibrios.
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  • 76
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    Archives of microbiology 126 (1980), S. 277-283 
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    Keywords: Bacteriophage ; Myxococcus ; λ ; Superooiled DNA ; Cross-linking ; Electron microscopy
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    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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  • 77
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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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    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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  • 78
    ISSN: 1432-072X
    Keywords: Ribulose bisphosphate carboxylase ; Quaternary structure ; Molecular weight ; Electron microscopy ; Cyanobacteria ; Synechococcus
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    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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  • 79
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    Archives of microbiology 135 (1983), S. 25-29 
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    Keywords: Electron microscopy ; Euglena mutabilis ; Flagellate ; Photomovement ; Photoreceptor ; Phototaxis ; Single-cell analysis ; Videomicroscopy
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    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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  • 80
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    Archives of microbiology 135 (1983), S. 169-175 
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    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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  • 81
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    Archives of microbiology 133 (1982), S. 97-99 
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    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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  • 82
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    Archives of microbiology 106 (1975), S. 195-200 
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    Keywords: Trichophyton terrestre ; Trichophyton rubrum ; Hyphal fusions ; Origin of intra-hyphal hyphae ; Electron microscopy
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    Notes: Abstract A cell observation chamber was designed to perform continuous photomicroscopic observations of hyphal anastomosis and the origin of intra-hyphal hyphae in Trichophyton terrestre and T. rubrum. These data were correlated with ultrastructural features of intra-hyphal hyphae. Hyphal fusions occurred commonly in either species of Trichophyton when incubated alone. In T. terrestre, empty hyphal segments adjoined by live units were invaded at the septa from both directions by new hyphal ingrowth. Continuous observations revealed that the intra-hyphal hyphae subsequently anastomosed via a lateral fusion peg. Similar intra-hyphal hyphae were shown in T. rubrum. Electron microscopic studies revealed ascomycetous septa in both conventional hyphae and intra-hyphal hyphae. For the latter, the cytoplasm and wall of the inner hypha were bounded by cytoplasmic organelles and another cell wall of the outer hypha.
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  • 83
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    Keywords: Thiobacillus ferrooxidans ; Sulfur production ; Sulfur oxidation ; Inhibitors ; Uncouplers ; Electron microscopy
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    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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  • 84
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    Keywords: Key wordsV. cholerae O139 ; Lipopolysaccharide ; Electron microscopy ; Freeze-substitution technique ; Capsule
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    Notes: Abstract The morphological and physical characteristics of the capsule of Vibrio cholerae O139 were examined. An electron microscopic study using the freeze-substitution technique showed that all of the V. cholerae strains of the O139 serogroup examined have a very thin fibrous layer on the outside of the outer membrane. In contrast, the mutants of strain O139, strain MO10T4 (which lacks capsule synthesis), and strain Bengal-2R1 (which fails to synthesize both the capsule and the O-antigen of lipopolysaccharide) were all found to have lost the surface layer. In addition, the capsule layer could also not be observed on the surface of V. cholerae strain O1. To determine the biological characteristics of the capsule of strains of the O139 serogroup, we investigated the serum killing activity and bacterial phagocytosis by polymorphonuclear leukocytes. The O139 strains were more resistant to the serum killing activity than were the V. cholerae O1 strain and the O139 mutant strains, thus suggesting that the existence of the capsule gave a serum-resistant character to the O139 strains. The surface character of the O139 strains had the same hydrophobic character as did that of the O139 mutant strains and the O1 strain. In addition, all the V. cholerae O1 and O139 strains examined, including the mutant strains, were effectively ingested by the human polymorphonuclear leukocytes. The number of ingested bacteria was not significantly different among the strains, and the ingestion of the acapsular O139 mutants thus showed that the capsule does not play an antiphagocytic role. These data suggest that the capsule of V. cholerae O139 has a physiological function different from that of the ordinal hydrophilic capsule that is found in invasive bacteria such as Klebsiella pneumoniae.
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  • 85
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    Keywords: Acetobacterium woodii ; Hydrogen-oxidizing acetate-forming anaerobe ; Fine structure ; Electron microscopy
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    Notes: Abstract Acetobacterium woodii is a Gram-positive anaerobic nonsporeforming bacterium able to grow on H2 and CO2 as sole sources of energy. The product of fermentation is acetic acid. Fine structural analysis showed rod-shaped flagellated cells, and coccoid cells without flagella arranged predominantly in pairs and chains. The cell wall was found to be composed of three layers. The cell surface exhibited a periodic array of particles consisting of subunits. The cytoplasmic membrane showed particles either either in random distribution or in a hexagonal pattern. Intracytoplasmic membranes were rarely observed, whereas inclusion bodies of varying shapes, predominantly in an uncommon disc-shape, could frequently be observed. Their content was dissolved in ultrathin sections indicating hydrophobic nature.
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  • 86
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    Archives of microbiology 119 (1978), S. 87-90 
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    Keywords: Salmonella typhimurium strain LT2 (ColIb) ; Cryptic plasmids ; Electron microscopy
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    Notes: Abstract Small cryptic plasmids of molecular weights ranging from 1 to 3 Mdal were detected by electron microscopy in Salmonella typhimurium strain LT2 (ColIb). They were divided into different size classes. Two of the cryptic plasmids were transferred simultaneously with ColIb to Escherichia coli.
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  • 87
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    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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  • 88
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    Archives of microbiology 130 (1981), S. 339-343 
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    Keywords: Leptospira ; Halophilic ; Electron microscopy ; X-ray analysis ; Inclusions ; Cytoplasmic tubules
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    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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  • 89
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    Archives of microbiology 129 (1981), S. 129-134 
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    Keywords: Cockroach ; Hindgut ; Distribution ; Microbial morphotypes ; Transmission ; Electron microscopy ; Statistical analysis ; Eublaberus posticus
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    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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  • 90
    ISSN: 1432-072X
    Keywords: Biological control ; Cyanobacteria ; Electron microscopy ; Entrapment ; lysis ; Myxococcus ; Phormidium ; Spherule
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    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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  • 91
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    Archives of microbiology 138 (1984), S. 273-277 
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    Keywords: Cyanobacteria ; Secondary metabolite ; Allelopathy ; Photosynthesis ; Electron transport ; Thylakoids ; Herbicides ; Electron microscopy
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    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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  • 92
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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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    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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  • 93
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    Archives of microbiology 142 (1985), S. 259-261 
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    Keywords: Methanogenic bacteria ; Plasmid isolation ; Alkaline lysis ; CsCl gradient ; Restriction endonuclease mapping ; Electron microscopy ; DNA homology
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    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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  • 94
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    Archives of microbiology 143 (1986), S. 400-402 
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    Keywords: E. coli relA +/relA ; Starvation survival ; Guanosine tetraphosphate ; Electron microscopy ; Glycogen
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    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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  • 95
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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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  • 96
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    Archives of microbiology 163 (1995), S. 300-306 
    ISSN: 1432-072X
    Keywords: Key wordsNitrosomonas europea ; Hydroxylamine ; oxidoreductase (HAO) ; Electron microscopy ; Electron ; spectroscopic imaging ; Quaternary structure
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    Notes: Abstract The hydroxylamine oxidoreductase from Nitrosomonas europaea was prepared to apparent electrophoretic homogeneity. Electron microscopy of negatively stained preparations of the sample revealed an overall diameter of about 8.8 nm of the enzyme particle. The native structure was determined as a tetrahedron-like assembly of identical subunits exhibiting four protein masses.
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  • 97
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    Archives of microbiology 146 (1986), S. 267-274 
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    Keywords: Photosynthesis ; Membrane structure ; Electron microscopy ; Ectothiorhodospira ; Serial thin sectioning ; Three dimensional reconstruction
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    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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  • 98
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    Keywords: Trichoderma reesei ; Xylanase ; Ultrastructural localization ; Immunogold labelling ; Electron microscopy
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    Notes: Abstract The intracellular location of the “low-molecular weight, alkaline” xylanase (XYN II) of Trichoderma reesei RUT C-30 was investigated during growth on xylan, using immunoelectron microscopy. A monoclonal antibody, produced against XYN II, was used for this purpose. The enzyme was found at the endoplasmic reticulum and in electron dense 0.2 to 0.8 μm vesicles, as well as in the vacuole, at the plasma membrane and in the fungal cell-wall. No staining occured in the cytoplasm, the mitochondria and the nucleus. No Golgi-like structures could be seen. Addition of the carboxylic ionophore monensin blocked xylanase as well as total protein secretion. The results are discussed with respect to XYN II being secreted by T. reesei via a pathway involving the endoplasmic reticulum and secretory vesicles and/or the vacuole.
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  • 99
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    Keywords: Malonomonas rubra ; Propionigenium modestum ; Malonate decarboxylase ; Methylmalonyl-CoA decarboxylase ; Biotin ; Avidin ; Electron microscopy ; High pressure freezing ; Immunolabeling
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Malonate decarboxylase of Malonomonas rubra is a complex enzyme system involving cytoplasmic and membrane-bound components. One of these is a biotin-containing protein of Mr 120'000, the location of which in the cytoplasm was deduced from the following criteria: (i) If the cytoplasm was incubated with avidin and the malonate decarboxylase subsequently completed with the membrane fraction the decarboxylase activity was abolished. The corresponding incubation of the membrane with avidin, however, was without effect. (ii) Western blot analysis identified the single biotin-containing polypeptide of Mr 120'000 within the cytoplasm. (iii) Transmission electron micrographs of immuno-gold labeled M. rubra cells clearly showed the location of the biotinyl protein within the cytoplasm, whereas the same procedure with Propionigenium modestum cells indicated the location of the biotin enzyme methylmalonyl-CoA decarboxylase in the cell membrane. The biotin-containing protein of the M. rubra malonate decarboxylase enzyme system was not retained by monomeric avidin-Sepharose columns but could be isolated with this column in a catalytically inactive form in the presence of detergents. If the high binding affinity of tetrameric avidin towards biotin was reduced by destructing part of the tryptophan residues by irradiation or oxidation with periodate, the inhibition of malonate decarboxylase by the modified avidin was partially reversed with an excess of biotin. Attempts to purify the biotin protein in its catalytically active state using modified avidin columns were without success.
    Type of Medium: Electronic Resource
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  • 100
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 160 (1993), S. 206-213 
    ISSN: 1432-072X
    Keywords: Treponema denticola ; Spirochetes ; Ultrastructure ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The formation of quasi-multicellular bodies of Treponema denticola was analysed using different electron microscopical methods. These bacteria could develop four different conformations: (i) normal helical forms; (ii) twisted spirochetes, forming plaits; (iii) twisted spirochetes, forming club-like structures; (iv) spherical bodies in different size. Treponemes within spherical bodies, plaits, and clubs proved to be enclosed in a common outer sheath in which the normal arrangement of their axial flagella was lost. The development of the quasi-multicellular bodies starting from the monoforme spirochetes was elucidated and this morphogenetic process is illustrated by a schematic drawing. Factors which might be involved in the induction of the structures are discussed and their possible pathogenetic importance is considered.
    Type of Medium: Electronic Resource
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