scholarly journals FERRITIN IN THE FUNGUS PHYCOMYCES

1971 ◽  
Vol 48 (1) ◽  
pp. 15-28 ◽  
Author(s):  
Charles N. David ◽  
Kenneth Easterbrook
Keyword(s):  
Lipid Droplets ◽  
Developmental Stages ◽  
Buoyant Density ◽  
Sedimentation Coefficient ◽  
Fold Increase ◽  
Iron Level ◽  
Phycomyces Blakesleeanus ◽  
Thin Sections ◽  
Iron Protein ◽  
Ferritin Iron

The iron-protein ferritin has been purified from mycelium, sporangiophores, and spores of the fungus Phycomyces blakesleeanus. It has a protein-to-iron ratio of 5, a sedimentation coefficient of 55S, a buoyant density in CsCl of 1.82 g/cm3, and the characteristic morphology of ferritin in the electron microscope. Apoferritin prepared from Phycomyces ferritin has a sedimentation coefficient of 18S and consists of subunits of molecular weight 25,000. In the cytoplasm of Phycomyces, ferritin is located on the surface of lipid droplets (0.5–2.0 µ in diameter) where it forms crystalline monolayers which are conspicuous in electron micrographs of sporangiophore thin-sections. Ferritin is found in all developmental stages of Phycomyces but is concentrated in spores. The level of ferritin iron is regulated by the iron level in the growth medium, a 50-fold increase occurring on iron-supplemented medium.

Ultrastructure and morphogenesis of the synnema of Ceratocystis ulmi

1973 ◽  
Vol 51 (9) ◽  
pp. 1565-1571 ◽  
Author(s):  
James L. Harris ◽  
Willard A. Taber
Keyword(s):  
Electron Microscopy ◽  
Bark Beetles ◽  
Lipid Droplets ◽  
Developmental Stages ◽  
Dutch Elm Disease ◽  
Thin Sections ◽  
Transmission Electron ◽  
Distinctive Structure ◽  
Elm Bark Beetles ◽  
Scanning Electron

Ceratocystis ulmi, the Ascomycete responsible for Dutch elm disease, may sporulate by means of a distinctive structure, the synnema, common in nature in the tunnels of elm bark beetles. Developmental stages of this structure and its spores were examined by light microscopy, by conventional transmission electron microscopy of thin sections and freeze-etch replicas, and by scanning electron microscopy of whole spores and fruiting structures. The synnema is a tight bundle of darkly pigmented hyphae growing erect from the substrate and terminated by a mass of colorless, wet spores. A layer of slime covering the hyphal bundle and random cross-connections between parallel hyphae appear to function in stabilization of the structure. Organelles typical of Ascomycetes fill the spores, vegetative hyphae, and young synnemal hyphae. However, in mature synnemal hyphae, the cytoplasmic contents degenerate, leaving only membranous vesicles in the partially collapsed hyphae. The spores contain large lipid droplets not found in either vegetative or synnemal hyphae indicating some differences in metabolism of spores and hyphae. Most synnemal spores form on sympodulae, but some spores form on intrahyphal hyphae.

The ultrastructure of oogonia and oocytes in the foetal and neonatal rat

1962 ◽  
Vol 157 (966) ◽  
pp. 99-114 ◽  
Keyword(s):  
Meiotic Prophase ◽  
Developmental Stages ◽  
Post Partum ◽  
Chromatin Condensation ◽  
Complex Form ◽  
Neonatal Rat ◽  
Mitotic Chromosomes ◽  
Thin Sections ◽  
Cytoplasmic Organelles ◽  
Spindle Fibres

Ovaries from eighty foetal and neonatal rats (aged 16·0 days post coitum to 4 days post partum ) were examined under the electron microscope. All the normal developmental stages (oogonia and oocytes in the leptotene, zygotene, pachytene, diplotene and dictyate phases of meiotic prophase) were identified. Patterns of degeneration (‘atretic divisions’, ‘ Z ’ cells and atresia at the diplotene phase), whose histological appearance and incidence have been recorded by Beaumont & Mandl (1962), were also recognized. The nuclei of oocytes at the leptotene phase contain single electron dense threads which become aligned in parallel pairs at the following phase (zygotene). A third finer fibril half-way between them appears at pachytene (tripartite ribbon). The longitudinal segments of threads, visible in ultra-thin sections, become shorter, presumably due to coiling. Nuclei at the diplotene phase contain single threads essentially similar to those seen at leptotene. As the oocyte enters the dictyate or resting phase, electron dense threads disappear from the nucleus. These observations suggest that the threads represent chromosomal ‘cores’. Nucleolus-like components persist throughout meiotic prophase, and at the diplotene phase regain the more complex form typical of oogonia. The cytoplasmic organelles become more numerous and complex as the oocyte approaches the dictyate phase. ‘Atretic divisions’ in oogonia are characterized by the persistence of long segments of nuclear membrane and the appearance of vesicles enveloped by a double membrane resembling the nuclear envelope. The dense masses of ‘chromatin’ (mitotic chromosomes) are more rounded than in normal cells at metaphase, and tend to coalesce. Spindle fibres have not been observed. Cytoplasmic organelles tend to increase in number and complexity. ‘ Z ’ cells (cells degenerating largely at the pachytene phase) show heavy ‘chromatin’ condensation around the tripartite ribbons. The major cytoplasmic changes consist in swelling of the endoplasmic reticulum, vacuolation of mitochondria and increase in incidence of multilamellar bodies. Atretic oocytes at the diplotene phase differ markedly from ‘ Z ’ cells in that ‘chromatin’ condensation around electron dense threads (single) is markedly less prominent. Cytoplasmic changes are similar to those of ‘ Z ’ cells, but also include a rise in the incidence of multivesicular and other complex bodies. All three types of degenerating cells are removed from the ovary by the phagocytic activity of neighbouring somatic cells.

scholarly journals The Isolation and Oharacterization of An Rna Bacteriophage

1964 ◽  
Vol 17 (3) ◽  
pp. 719 ◽  
Author(s):  
CI Davern
Keyword(s):  
Escherichia Coli ◽  
Dna Synthesis ◽  
Culture Medium ◽  
Buoyant Density ◽  
Sedimentation Coefficient ◽  
Caesium Chloride ◽  
Specific Phage ◽  
K 12 ◽  
Male Specific ◽  
Rna Phage

An enrichment procedure for the isolation of RNA bacteriophage is described. The method involves the inoculation of sewage samples into cultures of Escherichia coli K-12 Hfr under conditions where DNA synthesis is restricted by the addition of 5-fiuorodeoxyuridine to the culture medium. Six phage isolates were made and all of them were shown to be male-specific. One of the male-specific phage was further characterized as an RNA phage, having very similar properties to RNA phage already isolated in other parts oftha world. This RNA phage has a buoyant density of 1�42 g/cm3 in caesium chloride. and has a sedimentation coefficient of 79'5 Sin O'Ol:M Tria-HOI buffer, pH 7� 4, at 20�0.

Disappearance of the covalently closed circular cryptic plasmid PO-2 in Salmonella pullorum MS35 containing F77

1974 ◽  
Vol 20 (4) ◽  
pp. 551-557
Author(s):  
Paul W. Stiffler ◽  
D. E. Schoenhard
Keyword(s):  
Salmonella Typhimurium ◽  
Sucrose Gradient ◽  
Buoyant Density ◽  
Sedimentation Coefficient ◽  
Apparent Effect ◽  
Donor Property ◽  
Circular Dna ◽  
Salmonella Pullorum ◽  
Cryptic Plasmids ◽  
Equilibrium Centrifugation

The physical basis of the donor property of Salmonella pullorum donor strains MS8300, MS830, and MS831 carrying the F77 factor from Salmonella typhimurium was investigated by dye-buoyant density equilibrium centrifugation and zonal centrifugation in neutral sucrose gradients. Centrifugation of the MS8300 and MS831 closed circular DNA material in a 20 to 31% neutral sucrose gradient resulted in a profile having one sharp peak of radioactivity with a sedimentation coefficient of 17 S and a broad peak extending from 65 to 70 S. The 17- and 65-S species were isolated from the isogenic F− strain MS83. These appeared identical with those isolated previously in S. pullorum MS53 as the cryptic plasmids PO-1 and PO-2 respectively. Cosedimentation of differentially labeled F77 DNA and the lysate containing the 65-S and 70-S species suggested that the 70-S species is the autonomous F77 factor in strains MS8300 and MS831. Lysates of MS830 similarly treated produced a profile containing the 17-S molecule and possibly some 70-S molecules but no 65-S molecules. It was concluded that the F77 factor was integrated in most cells and that the covalently closed circular state of PO-2 plasmid was lost. The mutation in the cysE gene of the F77 factor carried by MS831 had no apparent effect on the covalently closed circular nature of PO-2 plasmid, although F77 no longer seemed to mobilize the chromosome from the cysE locus.

On Colouring Epon-Embedded Tissue Sections with Sudan Black B or Nile Blue A for Light Microscopy

1963 ◽  
Vol s3-104 (65) ◽  
pp. 109-115
Author(s):  
S. M. McGEE-RUSSELL ◽  
N. B. SMALE
Keyword(s):  
Lipid Droplets ◽  
Nile Blue ◽  
Thin Sections ◽  
Nile Blue A ◽  
Sudan Black B ◽  
Glass Slides ◽  
Acetone Water ◽  
Negative Effect ◽  
Blue Background ◽  
Dark Blue

Sections of osmium-fixed tissues embedded in epon 812 colour with either Sudan black B or Nile blue A solutions to reveal a variety of detail by direct microscopy with normal apochromatic or semi-apochromatic objectives. The clarity of the coloration gives a picture fully comparable to that seen by phase-contrast microscopy. The plastic is not removed, 1-µ sections or thinner sections down to green or gold, are mounted on clean glass slides by drying down from 20% acetone/water after flattening on a hot plate. Colouring is carried out at room temperature in Sudan black B (saturated solution in 70% alcohol) for 1 to 2 h. The result is a reversed or negative effect, for the epon plastic takes the stain avidly, but dense elements of the tissue do not, and appear white against a blue background of stained plastic. Lipid droplets retain a capacity to colour, becoming dark blue to blue-black. Nile blue sulphate ( 1% aqueous solution colours thin sections of tissue in 1 to 2 h at 60° C, acting apparently as a basic dye on most cell elements, and also colours lipid droplets dark blue. After both techniques the sections are mounted in Farrants's medium.

Self-association, cooperativity and supercooperativity of oxygen binding by hemoglobins.

1998 ◽  
Vol 201 (8) ◽  
pp. 1073-1084 ◽  
Author(s):  
A F Riggs
Keyword(s):  
Muscle Tissue ◽  
Buoyant Density ◽  
Fold Increase ◽  
Heme Iron ◽  
Common Mechanism ◽  
Oxygen Binding ◽  
Hypoxic Environment ◽  
Self Association ◽  
The Stability ◽  
O2 Binding

Cooperative ligand binding by tetrameric vertebrate hemoglobins (Hbs) makes possible the delivery of oxygen at higher pressures than would otherwise occur. This cooperativity depends on changes in dimer-dimer interactions within the tetramer and is reflected in a 50 000-fold increase in the tetramer-dimer dissociation constant in human Hb upon oxygenation at pH 7.4, from approximately 2x10(-11)mol l-1 to approximately 10(-6)mol l-1. Hbs that undergo such ligand-dependent changes in association are widespread in non-vertebrates, where the mechanisms are very different from those in vertebrates. Oligomeric Hbs have been identified in organisms in five phyla (molluscs, echinoderms, annelids, phoronids and chordates) that dissociate to subunits upon oxidation of the heme iron and reassociate with the binding of ferric iron ligands such as CN-, N3- or NO2-. Thus, the valence and ligand state of the heme iron control the stability of a critical subunit interface. The broad distribution of this phenomenon suggests a common mechanism of communication between heme and interface that may be almost universal among non-vertebrate Hbs. This interaction may be similar to that known for the homodimeric Hb of the mollusc Scapharca inaequivalvis. Although muscle tissue Hbs or myoglobins (Mbs) are usually monomeric, with non-cooperative O2 binding, the radular muscles of gastropod molluscs and chitons have homodimeric Mbs that bind O2 cooperatively. Cooperative non-muscle tissue Hbs have also been identified. These include the neural Hb of the nemertean worm Cerebratulus lacteus and the Hb of the diving beetle Anisops assimilis, which exhibit deoxygenation-dependent self-association of monomers that is associated with high Hill coefficients. Calculations suggest that the 2-3 mmol l-1 concentration of Hb on a heme basis in the brain of Cerebratulus should substantially extend the time as an active predator in an anaerobic or hypoxic environment. Oxygen from the Hb of Anisops is delivered to a gas bubble and thereby controls the buoyant density. Many Hbs of amphibians, reptiles, birds and some embryonic mammals exhibit a further 'supercooperativity' of O2 binding which depends on reversible deoxygenation-dependent tetramer-tetramer association to form an assemblage with a very low affinity for O2. This phenomenon results in steeper O2-binding curves than exhibited by tetramers alone. The increased cooperativity should result in an increase in the amount of O2 delivered to the tissues and should be especially valuable for avian flight muscles.

A NON-NUCLEOTIDE POLYMER FOUND IN THE DNA OF THE SAND DOLLAR, ECHINARACHNIUS PARMA: II. PRELIMINARY CHARACTERIZATION

1967 ◽  
Vol 45 (2) ◽  
pp. 281-287 ◽  
Author(s):  
Herbert S. Rosenkranz
Keyword(s):  
Buoyant Density ◽  
Sedimentation Coefficient ◽  
Negative Electrode ◽  
Cesium Chloride ◽  
Amino Sugar ◽  
Electrophoretic Analysis ◽  
Positive Test ◽  
Preliminary Characterization ◽  
Acid Hydrolysate

A preliminary characterization of the non-nucleotidic component present in the DNA of Echinarachnius parma was undertaken. This material has an extremely high sedimentation coefficient (907 S). It contains no deoxyribose and presumably no ribose. After acid hydrolysis it was strongly ninhydrin-positive and also gave positive tests for reducing sugars as well as a slightly positive test for amino sugars. Upon electrophoretic analysis of an acid hydrolysate, three ninhydrinpositive spots were detected. One of these migrated to the negative electrode with a mobility identical with that of galactosamine, the other migrated to the positive electrode, and the third was neutral at pH 6.3. The spot with a mobility identical with that of galactosamine also gave a positive test for amino sugar. The material was not attacked by α-amylase. However, digestion with a crude trypsin preparation resulted in loss of the banding property in gradients of cesium chloride. Exposure to purified trypsin did not completely digest it, but caused an increase in buoyant density.

155 CHARACTERIZATION OF LIPID DROPLETS IN BOS INDICUS AND BOS TAURUS EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 226 ◽  
Author(s):  
E. P. López-Damián ◽  
T. Fiordelisio ◽  
M. A. Lammoglia ◽  
M. Alarcón ◽  
M. Asprón ◽  
...  
Keyword(s):  
Lipid Droplets ◽  
Embryo Quality ◽  
Developmental Stages ◽  
Bos Taurus ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Bos Indicus ◽  
Nile Red ◽  
Blastocyst Stage ◽  
Transfer Program

Accurate evaluation of bovine embryos for assessing developmental stage and quality is critical to the success of any embryo transfer program. However, this evaluation process has been reported to be highly subjective in Bos indicus (BI) and can vary as much as 23% compared with that of Bos taurus (BT). These differences in assessment may be related to the quantity of lipid droplets (LD) within the embryo, which has been shown to have a negative effect in cryopreserving embryos. The aim of the present study was to characterize the number and size of LD in different developmental stages of fresh embryos from BI and BT and to compare LD across the three different embryo quality grades (1 = excellent or good, 2 = fair, and 3 = poor). Nonsurgical embryo collection was performed 7 days post-insemination in 10 BI and 10 BT females. Forty-eight embryos were evaluated for stage and grade using stereoscopic microscopy, processed for transmission electron microscopy, and stained with Nile red. Digitalized images were analyzed with ImageJ (National Institutes of Health, Bethesda, MD, USA), contour of lipid droplets were designed, and values of perimeter, area, and fluorescence intensity were assessed. Nonparametric statistical analysis (Mann–Whitney) was utilized. There was no difference in LD number for BT or BI for morulae and blastocyst; however, BI morulae presented larger LD compared with blastocyst stage embryos (286 µm2 v. 223 µm2; P < 0.05). Likewise, BI TF cells had more LD compared with inner cell mass (ICM) cells (48 v. 36; P < 0.05). BT TF cells exhibited larger LD compared with ICM cells (149 µm2 v. 128 µm2; P < 0.05), while BI embryos exhibited a larger area of LD in the ICM compared with the TF (591 µm2 v. 472 µm2; P < 0.05). In all embryos, BI contained more lipid droplets than BT (78 v. 49; P < 0.05). Across all quality grades (good, fair, and poor) there was no difference in the number of LD in BT embryos; however, BI grade-3 embryos presented more LD than grade-1 (36 v. 25). BT embryos LD were larger than BI LD (907 µm2 v. 625 µm2; P < 0.05). Fluorescence images showed higher arbitrary units of fluorescence (auf) for LD in BI. Compared with BT embryos (386 auf v. 280 auf; P < 0.05). These results suggest that BI embryos contain more and smaller LD than BT embryos and the LD described for BI embryo quality grade 1 are larger than those of quality grades 2 and 3, and even though the number of LD in morulae and blastocyst stage embryos are not different LD size is reduced as development occurs. Research funding provided by UNAM-DGAPA-PAPIIT IN200810.

Characteristics of calcium transport and binding by rat myometrium plasma membrane subfractions

1980 ◽  
Vol 239 (3) ◽  
pp. C66-C74 ◽  
Author(s):  
A. K. Grover ◽  
C. Y. Kwan ◽  
J. Crankshaw ◽  
D. J. Crankshaw ◽  
R. E. Garfield ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cytochrome C ◽  
Buoyant Density ◽  
Membrane Vesicles ◽  
Ionophore A23187 ◽  
Plasma Membrane Vesicles ◽  
Thin Sections ◽  
Cytochrome C Reductase ◽  
Ca Uptake ◽  
High Concentrations

A gradient has been designed to yield two subfractions of plasma membrane vesicles from rat myometrium, a low buoyant density (8-24% sucrose) fraction N1 richer in 5'-nucleotidase and a higher buoyant density (24-30% sucrose) fraction N2, instead of a previously described fraction F1. Both N1 and N2 had very low activities of NADPH-cytochrome c reductase and succinate-cytochrome c reductase. Electron micrographs of thin sections of N1 showed clear vesicles, whereas N2 consisted of vesicles with electron-dense bodies attached to them. These plasma membrane vesicles can actively take up Ca. The active uptake of Ca was potentiated by oxalate and phosphate and abolished by the Ca ionophore A23187. Dilution of actively loaded vesicles in isotonic media containing EGTA led to loss of a small proportion of the stored Ca instantaneously and the remainder more slowly in a biphasic manner. Dilution in hypotonic media with EGTA led to a release of a much larger proportion of the accumulated Ca. A23187 at high concentrations (10 microM) caused a release of all the sequestered Ca whether the active Ca uptake had been carried out in the presence or in the absence of oxalate. A23187, 0.5 microM, released all the sequestered Ca from the vesicles that were actively loaded in the absence of oxalate, but only 37% when the vesicles were actively loaded with Ca in the presence of oxalate. Comparison of the composite plasma membrane fraction F1 (8-30% sucrose) and the subfractions N1 and N2 showed that they had different capacities for Ca uptake in the presence and absence of ATP. An attempt has been made to analyze the active Ca-uptake data in terms of various Ca pools.

Die Bildung von RNS-Doppelstrang zur Vermehrung eines RNS enthaltenden Bakteriophagen

1964 ◽  
Vol 19 (7) ◽  
pp. 593-604 ◽  
Author(s):  
Hans Christian Kaerner ◽  
Hartmut Hoffmann-Berling
Keyword(s):  
Molecular Weight ◽  
Buoyant Density ◽  
Sedimentation Coefficient ◽  
Radioactive Material ◽  
Double Stranded Rna ◽  
Infected Cells ◽  
Rna Phage ◽  
Denaturation Profile ◽  
Molecular Weight Form ◽  
Latent Phase

The RNA phage fr induces in Escherichia coli cells the production of double stranded RNA, which is identified by its thermal denaturation profile ( Tm 101 °C in 0,2-m. Na⊕ ), by its nonreactivity with formaldehyde and by its buoyant density in Cs2SO4 (1,609 g cm-3 , compared to that of fr-RNA = 1,634 g cm-3 ). Unless denatured the double strand is resistant to RNase. In its high molecular weight form the double stranded R N A has twice the weight of fr-RNA, as estimated from the sedimentation coefficient (s20 = 14,5). The base ratios are those expected for a double stranded replicative from of fr-RNA. By melting and annealing one of the strands of the non-radioactive material can be exchanged for 32P-fr-RNA from phage particles. Infectiosity of the doublestranded RNAhas not yet been shown. Extracts from infected cells contain double strand bound to the 30 - 50 s fraction; there is also double strand in the supernatant, apparently in the form of relatively low molecular weight fragments. The double stranded RNA, isolated at the height of infection, accounts for 3 - 8% of the cellular RNA. Cells infected with 32P-fr show a surprisingly large part of the infecting RNA bound to ribosomes quite late in the latent phase. The meaning of this result is discussed.

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