Metabolic development and mitochondrial changes during cyclopiazonic acid production in Penicillium cyclopium

1977 ◽  
Vol 23 (7) ◽  
pp. 856-872 ◽  
Author(s):  
Dianne C. Neethling ◽  
Robert M. McGrath
Keyword(s):  
Secondary Metabolism ◽  
Mevalonic Acid ◽  
Cyclopiazonic Acid ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Rate Limiting Step ◽  
Penicillium Cyclopium ◽  
Intermediate Time ◽  
Atp Generation ◽  
Rate Limiting

During a study of cyclopiazonic acid (total α- and β-cyclopiazonic acids, CA) production by Penicillium cyclopium Westling, it was found that nitrogen depletion controlled the advent of secondary metabolism and that nitrate reduction was not rate-limiting. In concert with N depletion both tryptophan (Trp) biosynthesis and glucose catabolism fell. The rate of Trp production dropped to match the rate of CA biosynthesis. Glucose utilization varied between an early and late preference for the pentose cycle, with a mixture of pentose and Emden–Meyerhof pathways at an intermediate time. This pattern may well reflect the need for erythrose-4-phosphate in Trp biosynthesis. Additions of Trp caused a drop in CA production possibly because of repression of Trp synthetase. Exogeneous dimethylallylpyrophosphate (DMAPP) increased CA biosynthesis, while mevalonic acid had no effect which suggested a rate-limiting step between these two metabolites. These changes probably best fit Bu'Lock's hypothesis that secondary metabolism links fermentative and aerobic metabolism (ATP generation). In this case protein and polyisoprenoid biosynthesis, sugar utilization, and energy requirements would all be interlocked with the advent of secondary metabolism. The possible relationship between the observed metabolic changes and energy control were supported by electron-microscopic examination of the mitochondria in situ, when these ATP-generating organelles underwent major changes. Addition of ATP to the culture depressed CA production, as did excessive aeration. The metabolic regulator cAMP had no effect other than to increase autocatalysis.

The effects of tunicamycin, mevinolin and mevalonic acid on HMG-CoA reductase activity and nuclear division in the myxomycete Physarum polycephalum

1989 ◽  
Vol 92 (3) ◽  
pp. 341-344
Author(s):  
W. Engstrom ◽  
O. Larsson ◽  
W. Sachsenmaier
Keyword(s):  
Physarum Polycephalum ◽  
Nuclear Division ◽  
Reductase Activity ◽  
Mevalonic Acid ◽  
Minor Effect ◽  
Hmg Coa Reductase ◽  
Rate Limiting Step ◽  
Coa Reductase ◽  
A Minor ◽  
Rate Limiting

The effects of two inhibitors of 3-hydroxy 3-methyl glutaryl-coenzyme A reductase (tunicamycin and mevinolin) on nuclear division in the myxomycete Physarum polycephalum were examined. Tunicamycin exerted a minor effect on division in synchronized cultures, whereas mevinolin delayed the second, third and fourth nuclear divisions with increasing efficiency. Mevinolin also appeared to be the more potent inhibitor of HMG-CoA reductase, which catalyses the rate-limiting step in the biosynthesis of cholesterol and other isoprene derivatives. These effects of mevinolin could be partially reversed by the addition of mevalonate, suggesting that mevinolin exerts its inhibitory effects on Physarum nuclear division by decreasing the activity of HMG-CoA reductase.

ACETATE INCORPORATION INTO CHOLESTEROL AND FATTY ACIDS BY LIVER SLICES FROM RATS FED COMMERCIAL OR SEMISYNTHETIC DIETS: THE EFFECT OF DIETARY FATS

1964 ◽  
Vol 42 (1) ◽  
pp. 71-78 ◽  
Author(s):  
K. K. Carroll
Keyword(s):  
Fatty Acids ◽  
Mevalonic Acid ◽  
Dietary Fats ◽  
Acetate Incorporation ◽  
Commercial Diet ◽  
Rate Limiting Step ◽  
Liver Slices ◽  
Cholesterol Biosynthetic Pathway ◽  
Rate Limiting ◽  
Insoluble Portion

Liver slices from rats fed a commercial diet incorporated more acetate-1-C14 into cholesterol than did slices from rats fed semisynthetic diets. The stimulatory effect of the commercial diet was due in part to a saponifiable component of the ether-soluble portion of the diet, possibly linoleic acid; and in part to the ether-insoluble portion of the diet. Dietary fats stimulated acetate incorporation into cholesterol more when mixed with the ether-insoluble portion of commercial diet than when mixed with semisynthetic diet. With either diet, unsaturated fats stimulated incorporation more than saturated fats. The rate-limiting step in these experiments was prior to mevalonic acid in the cholesterol biosynthetic pathway. Acetate incorporation into fatty acids was little affected by the nature of the dietary fat or the diet in which it was fed.

Electron microscopic studies on ultrathin frozen sections of lactating mouse mammary gland

1978 ◽  
Vol 36 (2) ◽  
pp. 46-47
Author(s):  
Jan Zarzycki ◽  
Joseph Szroeder
Keyword(s):  
Mammary Gland ◽  
Protein Denaturation ◽  
Chemical Constituents ◽  
Freeze Substitution ◽  
Electron Microscopic Examination ◽  
Mouse Mammary Gland ◽  
Electron Microscopic ◽  
Preparation Methods ◽  
Microscopic Studies ◽  
Ultrathin Frozen Sections

The mammary gland ultrastructure in various functional states is the object of our investigations. The material prepared for electron microscopic examination by the conventional chemical methods has several limitations, the most important are the protein denaturation processes and the loss of large amounts of chemical constituents from the cells. In relevance to this,one can't be sure about a degree the observed images are adequate to the realy ultrastructure of a living cell. To avoid the disadvantages of the chemical preparation methods,some autors worked out alternative physical methods based on tissue freezing / freeze-drying, freeze-substitution, freeze-eatching techniqs/; actually the technique of cryoultraraicrotomy,i,e.cutting ultrathin sections from deep frozen specimens is assented as a complete alternative method. According to the limitations of the routine plastic embbeding methods we were interested to analize the mammary gland ultrastructure during lactation by the cryoultramicrotomy method.

A Comparative Study of Fractographic Replicas

1974 ◽  
Vol 32 ◽  
pp. 566-567
Author(s):  
Loren Anderson ◽  
Pat Pizzo ◽  
Glen Haydon
Keyword(s):  
Electron Microscopy ◽  
Electron Microscopic Examination ◽  
Direct Examination ◽  
Electron Microscopic ◽  
Reliable Technique ◽  
Service Failures ◽  
Transmission Electron ◽  
Mode Of Failure ◽  
Scanning Electron Microscopic ◽  
Scanning Electron

Transmission electron microscopy of replicas has long been used to study the fracture surfaces of components which fail in service. Recently, the scanning electron microscope (SEM) has gained popularity because it allows direct examination of the fracture surface. However, the somewhat lower resolution of the SEM coupled with a restriction on the sample size has served to limit the use of this instrument in investigating in-service failures. It is the intent of this paper to show that scanning electron microscopic examination of conventional negative replicas can be a convenient and reliable technique for determining mode of failure.

Novel methods for demonstrating osseointegration of hydroxylapatite-coated titanium hip prostheses

1991 ◽  
Vol 49 ◽  
pp. 34-35 ◽  
Author(s):  
P. Frayssinet ◽  
J. Hanker ◽  
D. Hardy ◽  
B. Giammara
Keyword(s):  
Hip Prosthesis ◽  
Ethanol Concentration ◽  
Bone Matrix ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Hip Prostheses ◽  
Cellular Inclusions ◽  
Microscopic Studies ◽  
Diamond Saw ◽  
Plasma Sprayed

Prostheses implanted in hard tissues cannot be processed for electron microscopic examination or microanalysis in the same way as those in other tissues. For these reasons, we have developed methods allowing light and electron microscopic studies as well as microanalysis of the interface between bone and a metal biomaterial coated by plasma-sprayed hydroxylapatite(HA) ceramic.An HA-coated titanium hip prosthesis (Corail, Landos, France), which had been implanted for two years, was removed after death (unrelated to the orthopaedic problem). After fixation it was dehydrated in solutions of increasing ethanol concentration prior to embedment in polymethylmethacrylate(PMMA). Transverse femur sections were obtained with a diamond saw and the sections then carefully ground to a thickness of 200 microns. Plastic-embedded sections were stained for calcium with a silver methenamine modification of the von Kossa method for calcium staining and coated by carbon. They have been examined by back-scatter SEM on an ISI-SS60 operated at 25 KV. EDAX has been done on cellular inclusions and extracellular bone matrix.

Properties of Isolated Mitochondria of Agaricus Bisporus: Their Relationship to Dormancy and the Germination of Spores

1973 ◽  
Vol 31 ◽  
pp. 458-459
Author(s):  
K. S. McCarty ◽  
R. F. Weave ◽  
L. Kemper ◽  
F. S. Vogel
Keyword(s):  
Mitochondrial Dna ◽  
Ethidium Bromide ◽  
Microscopic Examination ◽  
Agaricus Bisporus ◽  
Osmotic Shock ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Isolated Mitochondria ◽  
Dna Strands ◽  
Cytoplasmic Ribosomes

During the prodromal stages of sporulation in the Basidiomycete, Agaricus bisporus, mitochondria accumulate in the basidial cells, zygotes, in the gill tissues prior to entry of these mitochondria, together with two haploid nuclei and cytoplasmic ribosomes, into the exospores. The mitochondria contain prominent loci of DNA [Fig. 1]. A modified Kleinschmidt spread technique1 has been used to evaluate the DNA strands from purified whole mitochondria released by osmotic shock, mitochondrial DNA purified on CsCl gradients [density = 1.698 gms/cc], and DNA purified on ethidium bromide CsCl gradients. The DNA appeared as linear strands up to 25 u in length and circular forms 2.2-5.2 u in circumference. In specimens prepared by osmotic shock, many strands of DNA are apparently attached to membrane fragments [Fig. 2]. When mitochondria were ruptured in hypotonic sucrose and then fixed in glutaraldehyde, the ribosomes were released for electron microscopic examination.

Molecular Structure of the Outermost Cell Wall Component of Spirillum Serpens

1977 ◽  
Vol 35 ◽  
pp. 342-343
Author(s):  
Wah Chiu ◽  
David Grano
Keyword(s):  
Molecular Structure ◽  
Cell Wall ◽  
Outer Membrane ◽  
Gel Electrophoresis ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Cell Wall Component ◽  
Protein Components ◽  
Exposure Technique ◽  
Structural Aspects

The periodic structure external to the outer membrane of Spirillum serpens VHA has been isolated by similar procedures to those used by Buckmire and Murray (1). From SDS gel electrophoresis, we have found that the isolated fragments contain several protein components, and that the crystalline structure is composed of a glycoprotein component with a molecular weight of ∽ 140,000 daltons (2). Under an electron microscopic examination, we have visualized the hexagonally-packed glycoprotein subunits, as well as the bilayer profile of the outer membrane. In this paper, we will discuss some structural aspects of the crystalline glycoproteins, based on computer-reconstructed images of the external cell wall fragments.The specimens were prepared for electron microscopy in two ways: negatively stained with 1% PTA, and maintained in a frozen-hydrated state (3). The micrographs were taken with a JEM-100B electron microscope with a field emission gun. The minimum exposure technique was essential for imaging the frozen- hydrated specimens.

Electron Microscopic Examination of a Transplantable Rat Mammary Carcinoma

1968 ◽  
Vol 26 ◽  
pp. 20-21
Author(s):  
S. Shirahama ◽  
G. C. Engle ◽  
R. M. Dutcher
Keyword(s):  
Electron Microscope ◽  
Osmium Tetroxide ◽  
Electron Microscopic Examination ◽  
Undifferentiated Carcinoma ◽  
Uranyl Acetate ◽  
Sprague Dawley ◽  
Electron Microscopic ◽  
North West ◽  
X Irradiation ◽  
Tissue Specimens

A transplantable carcinoma was established in North West Sprague Dawley (NWSD) rats by use of X-irradiation by Engle and Spencer. The tumor was passaged through 63 generations over a period of 32 months. The original tumor, an adenocarcinoma, changed into an undifferentiated carcinoma following the 19th transplant. The tumor grew well in NWSD rats of either sex at various ages. It was invariably fatal, causing death of the host within 15 to 35 days following transplantation.Tumor, thymus, spleen, and plasma from 7 rats receiving transplants of tumor at 3 to 9 weeks of age were examined with an electron microscope at intervals of 8, 15, 22 and 30 days after transplantation. Four normal control rats of the same age were also examined. The tissues were fixed in glutaraldehyde, postfixed in osmium tetroxide and embedded in Epon. The plasma was separated from heparanized blood and processed as previously described for the tissue specimens. Sections were stained with uranyl acetate followed by lead citrate and examined with an RCA EMU-3G electron microscope.

Artifact-free electron microscopic immunocytochemistry on rat brain tissue

1991 ◽  
Vol 49 ◽  
pp. 272-273
Author(s):  
Veronika Burmeister ◽  
N. Ludvig ◽  
P.C. Jobe
Keyword(s):  
Microscopic Examination ◽  
Free Electron ◽  
Electron Microscopic Examination ◽  
Ultrastructural Localization ◽  
Rabbit Serum ◽  
Electron Microscopic ◽  
Electron Microscopic Immunocytochemistry ◽  
Phosphate Buffered Saline ◽  
Rat Brain Tissue ◽  
Immune Rabbit

Electron microscopic immunocytochemistry provides an important tool to determine the ultrastructural distribution of various molecules in both normal and pathologic tissues. However, the specific immunostaining may be obscured by artifactual immunoreaction product, misleading the investigator. Previous observations show that shortening the incubation period with the primary antibody from the generally used 12-24 hours to 1 hour substantially reduces the artifactual immunostaining. We now extend this finding by the demonstration of artifact-free ultrastructural localization of the Ca2/calmodulindependent cyclic nucleotide phosphodiesterase (CaM-dependent PDE) immunoreactivity in brain.Anesthetized rats were perfused transcardially with phosphate-buffered saline followed by a fixative containing paraformaldehyde (4%) and glutaraldehyde (0.25%) in PBS. The brains were removed, and 40μm sections were cut with a vibratome. The sections were processed for immunocytochemistry as described by Ludvig et al. Both non-immune rabbit serum and specific CaM-dependent PDE antibodies were used. In both experiments incubations were at one hour and overnight. The immunostained sections were processed for electron microscopic examination.

Some Effects of Oxidant Air Pollutants (Ozone and Peroxyacetyl Nitrate) on the Ultrastructure of Leaf Tissues

1972 ◽  
Vol 30 ◽  
pp. 360-361
Author(s):  
William W. Thomson ◽  
Elizabeth S. Swanson
Keyword(s):  
Air Pollutants ◽  
Electron Microscopic Examination ◽  
Peroxyacetyl Nitrate ◽  
Electron Microscopic ◽  
Growth Physiology ◽  
Physiology And Biochemistry ◽  
Entire Plant ◽  
Leaf Tissues ◽  
Gaseous Hydrocarbons ◽  
The Individual

The oxidant air pollutants, ozone and peroxyacetyl nitrate, are produced in the atmosphere through the interaction of light with nitrogen oxides and gaseous hydrocarbons. These oxidants are phytotoxicants and are known to deleteriously affect plant growth, physiology, and biochemistry. In many instances they induce changes which lead to the death of cells, tissues, organs, and frequently the entire plant. The most obvious damage and biochemical changes are generally observed with leaves.Electron microscopic examination of leaves from bean (Phaseolus vulgaris L.) tobacco (Nicotiana tabacum L.) and cotton (Gossipyum hirsutum L.) fumigated for .5 to 2 hours with 0.3 -1 ppm of the individual oxidants revealed that changes in the ultrastructure of the cells occurred in a sequential fashion with time following the fumigation period. Although occasional cells showed severe damage immediately after fumigation, the most obvious change was an enhanced clarity of the cell membranes.

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