96 results on '"Giometti, C S"'
Search Results
2. HIGH-RESOLUTION TWO-DIMENSIONAL ELECTROPHORETIC MAPPING OF HUMAN PROTEINS
- Author
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Anderson, N. L., primary, Edwards, J. J., additional, Giometti, C. S., additional, Willard, K. E., additional, Tollaksen, S. L., additional, Nance, S. L., additional, Hickman, B. J., additional, Taylor, J., additional, Coulter, B., additional, Scandora, A., additional, and Anderson, N. G., additional
- Published
- 1980
- Full Text
- View/download PDF
3. TWO-DIMENSIONAL ELECTROPHORESIS OF HUMAN SALIVA
- Author
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Giometti, C. S., primary and Anderson, N. G., additional
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- 1980
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- View/download PDF
4. Changes in muscle protein composition induced by disuse atrophy - Analysis by two-dimensional electrophoresis
- Author
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Ellis, S, Giometti, C. S, and Riley, D. A
- Subjects
Life Sciences (General) - Abstract
Using 320 g rats, a two-dimensional electrophoretic analysis of muscle proteins in the soleus and EDL muscles from hindlimbs maintained load-free for 10 days is performed. Statistical analysis of the two-dimensional patterns of control and suspended groups reveals more protein alteration in the soleus muscle, with 25 protein differences, than the EDL muscle, with 9 protein differences, as a result of atrophy. Most of the soleus differences reside in minor components. It is suggested that the EDL may also show alteration in its two-dimensional protein map, even though no significant atrophy occurred in muscle wet weight. It is cautioned that strict interpretation of data must take into account possible endocrine perturbations.
- Published
- 1985
5. XRF and XAFS Analysis of Electrophoretically Isolated Nondenatured Proteins
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Kemner, K. M., primary, Kelly, S. D., additional, O'Loughlin, E. J., additional, Khare, T., additional, Moe, L. A., additional, Fox, B. G., additional, Donnelly, M. I., additional, Londer, Y., additional, Schiffer, M., additional, and Giometti, C. S., additional
- Published
- 2005
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- View/download PDF
6. Identification of a heritable deficiency of the folate-dependent enzyme 10-formyltetrahydrofolate dehydrogenase in mice.
- Author
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Champion, K M, primary, Cook, R J, additional, Tollaksen, S L, additional, and Giometti, C S, additional
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- 1994
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7. Protein changes occurring during storage of platelet concentrates.
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Snyder, E. L., Dunn, B. E., Giometti, C. S., Napychank, P. A., Tandon, N. N., Ferri, P. M., and Hofmann, J. P.
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- 1987
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8. Human muscle proteins: analysis by two-dimensional electrophoresis.
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Giometti, C S, Danon, M J, and Anderson, N G
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- 1983
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9. Human muscle proteins.
- Author
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Giometti, C. S., Danon, M. J., and Anderson, N. G.
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- 1983
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10. Sequential muscle biopsy changes in a case of congenital myopathy.
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Danon, Moris J., Giometti, Carol S., Manaligod, Jose R., Swisher, Charles, Danon, M J, Giometti, C S, Manaligod, J R, and Swisher, C
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- 1997
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11. Shotgun Proteomics of Methanococcus jannaschii and Insights into Methanogenesis
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Zhu, W., Reich, C. I., Olsen, G. J., Giometti, C. S., and Yates, J. R., III
- Abstract
Methanococcus jannaschii is an autotrophic hyperthermophilic archaeon isolated from an oceanic hydrothermal vent. Its primary pathway for energy production is methanogenesis from H
2 and CO2 . High-throughput Multidimensional Protein Identification Technology based on microcapillary LC/LC/MS/MS was used to investigate the proteome of M. jannaschii and the methanogenesis pathway in cells grown in complex medium with high H2 supply. A total of 963 proteins have been unambiguously identified. The identified proteins represent ~54% of the whole genome of M. jannaschii. About 44% of the identified proteins are either conserved hypothetical or hypothetical proteins. We identified 83−95% of the proteins predicted to be involved in amino acid biosynthesis, cellular processes, central intermediary metabolism, energy metabolism, protein synthesis, transcription, and purine, pyridine, nucleoside, and nucleotide synthesis. Over 40% of these proteins have better than 50% sequence coverage. Approximately 90% of the predicted methanogenesis proteins were detected. In contrast, only 27−37% of predicted hypothetical proteins, proteins involved in transport and binding, and proteins with regulatory functions were identified. High peptide number, spectrum count, and sequence coverage have been used as indicators of high expression levels and are in good agreement with codon bias analysis. Predicted intein peptides were detected in MJ1043 (DNA-directed RNA polymerase, subunit A ), MJ0542 (phosphoenolpyruvate synthase), MJ0782 (transcription initiation factor IIB), and MJ1422 (putative replication factor C subunit). New peptides created by protein splicing were detected in MJ0885 (DNA dependent DNA polymerase), MJ0542, and MJ0782. The methanogenesis pathway and the enzymes involved are also discussed. Keywords: proteomics • MudPIT • LC/LC/MS/MS • Methanococcus jannaschii • mass spectrometry • SEQUEST • methanogenesis • archaea • intein- Published
- 2004
12. Gel electrophoresis for studying biological function
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Barany, M., Barany, K., and Giometti, C. S.
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- 1998
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13. Tropomyosin heterogeneity in human cells.
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Giometti, C S and Anderson, N L
- Abstract
Tropomyosin preparations from human platelets, human peripheral blood leukocytes from normal individuals and from a patient with chronic lymphocytic leukemia, human lymphoblastoid cells (GM607), human epithelial cells, and human skin fibroblasts have all been found to contain more than one protein when analyzed by two-dimensional gel electrophoresis. Although the lymphoid cell preparations consistently contain two proteins of almost identical molecular weight (Mr = 30,000), the platelet, epithelial cell, and fibroblast preparations contain two or more major proteins with molecular weights between 31,000 and 36,000, in addition to a major protein at 30,000. All of these proteins have characteristics in common with tropomyosin including slightly acidic isoelectric point (approximately pH 4), stability to heat and organic solvents, association with the cytoskeleton, and reactivity with antibody against skeletal muscle tropomyosin. The nonmuscle tropomyosin-like proteins were compared with tropomysins from human skeletal, cardiac, and smooth muscle by peptide mapping after partial proteolysis. The results showed one of the non-muscle proteins to be identical to the major smooth muscle tropomyosin in human uterus (myometrium) and another to be similar but not identical to skeletal muscle alpha-tropomyosin. The remainder of the proteins with tropomyosin characteristics was unique to non-muscle cells. In all, nine distinct human proteins with characteristics of tropomyosin are described. Charge variants of two of these proteins have been described previously.
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- 1984
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14. The fatty liver dystrophy mutant mouse: microvesicular steatosis associated with altered expression levels of peroxisome proliferator-regulated proteins.
- Author
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Rehnmark, S, Giometti, C S, Slavin, B G, Doolittle, M H, and Reue, K
- Abstract
Fatty liver dystrophy ( fld) is an autosomal recessive mutation in mice characterized by hypertriglyceridemia and fatty liver during neonatal development. The fatty liver in fld/fld mice spontaneously resolves between the age of 14-18 days, at which point the animals develop a neuropathy associated with abnormal myelin formation in peripheral nerve. We have investigated the morphological and biochemical alterations that occur in the fatty liver of neonatal fld/fld mice. Studies at the light and electron microscopic level demonstrated the accumulation of lipid droplets and hypertrophic parenchymal cells in fld neonates, with no apparent liver pathology after resolution of the fatty liver. To better characterize the biochemical basis for the development of fatty liver in fld mice, we compared protein expression patterns in the fatty liver of fld mice and in the liver of phenotypically normal (wild-type) littermates using quantitative two-dimensional gel electrophoresis. We detected 24 proteins with significantly altered expression levels (P < 0.001) in the fld fatty liver, 15 of which are proteins that are altered in abundance by peroxisome proliferating chemicals. As these compounds characteristically elicit changes in the expression of mitochondrial and peroxisomal enzymes involved in fatty acid oxidation, we quantitated rates of fatty acid oxidation in hepatocytes isolated from fld and wild-type mice. These studies revealed that hepatic fatty acid oxidation in fld neonates is reduced by 60% compared to wild-type littermates. In hepatocytes from adult fld mice that no longer exhibit a fatty liver, oxidation rates were similar to those in hepatocytes from age-matched wild-type mice. These findings indicate that altered expression of proteins involved in fatty acid oxidation is associated with triglyceride accumulation in the fld fatty liver.
- Published
- 1998
15. A heritable variant of mouse liver ornithine aminotransferase (EC 2.6.1.13) induced by ethylnitrosourea.
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Giometti, C S, Tollaksen, S L, Gemmell, M A, Burcham, J, and Peraino, C
- Abstract
A variant of ornithine aminotransferase (OAT, EC 2.6.1.13) has been detected in an offspring of a male mouse treated with ethylnitrosourea. The evidence presented to support the identification of the protein variant (ENU 2) as altered OAT includes (a) a corresponding 50% decrease in the abundance of a protein, located one charge unit basic to the variant, which comigrates on two-dimensional gel patterns with purified mouse liver OAT; (b) the binding of anti-rat-OAT antibody to the variant; (c) the increased abundance of the variant protein in the livers of mice fed a high protein diet (85% casein); and (d) purification of the variant through an OAT purification protocol.
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- 1988
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16. Evidence for regulatory genes on mouse chromosome 7 that affect the quantitative expression of proteins in the fetal and newborn liver.
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Giometti, C S, Gemmell, M A, Taylor, J, Tollaksen, S L, Angeletti, R, and Gluecksohn-Waelsch, S
- Abstract
A series of deletions around the albino locus on mouse chromosome 7 is believed to include one or more regulatory genes that control the activities of a cluster of liver enzymes. To further characterize the functions of this region of the mouse genome, we have used quantitative two-dimensional electrophoresis to analyze the effects of two of these deletions, c3H and c14CoS, on the expression of liver proteins. More than 400 distinct protein gene products were quantitated in livers from fetal and newborn wild-type homozygous (cch/cch), heterozygous (cch/c3H or cch/c14CoS), and deletion homozygous (c3H/c3H or c14CoS/c14CoS) mice. Livers of fetal and newborn c3H heterozygotes and homozygous wild-type littermates produced qualitatively identical protein patterns after two-dimensional electrophoresis. In livers of c3H homozygous fetuses, however, abnormal amounts (either increased or decreased relative to homozygous wild-type and heterozygous littermates) of 29 proteins were found. Twenty-eight of these 29 protein anomalies were also found in livers of newborn c3H homozygotes. Livers of fetal and newborn mice homozygous for the c14CoS deletion, which overlaps the c3H deletion and produces a similar phenotype, expressed normal amounts of these proteins. One of the 29 proteins (MSN807) has an amino-terminal sequence similar to a 23-kDa translationally controlled protein abundant in mouse erythroleukemia and sarcoma-180 cells. These results suggest that normal chromosome 7 contains genes, located within the region of the c3H but not the c14CoS deletion, that regulate the abundance of specific proteins in the liver. These proteins cannot be related to the phenotypic alterations shared by the c3H and c14CoS deletions.
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- 1992
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17. Detection of heritable mutations as quantitative changes in protein expression.
- Author
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Giometti, C S, Gemmell, M A, Nance, S L, Tollaksen, S L, and Taylor, J
- Abstract
A computerized search for the appearance of heritable mutations (as indicated by changes in protein expression) was conducted on three sets of mice, whose sires had been either untreated, exposed to 3 gray units of gamma radiation, or treated with 150 mg/kg ethylnitrosourea. Proteins from the livers of approximately 800 mice were separated by two-dimensional electrophoresis, and abundances were measured by using image analysis techniques. Heritable mutations were detected by the appearance of new proteins or by the quantitative decrease in abundance of normally occurring proteins. Measurements of the variability of the protein abundance indicate that at least 48 proteins are consistent enough to be used in searches when mutations are expected to result in a 50% reduction in the normal amount of protein. New proteins were found in four offspring from ethylnitrosourea-treated mice, and in each case a nearby spot was found to be significantly diminished. These mutations were confirmed in subsequent generations. A computer-assisted search detected three of these mutations on the basis of the abundance decrease alone. These results indicate that two-dimensional electrophoresis can be used to detect mutations reflected as quantitative changes in protein expression, provided that the proteins to be monitored are quantitatively stable when samples from different individuals are compared.
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- 1987
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18. Electrophoresis '79: Advanced Methods, Biochemical and Clinical Applications: Internal Charge Standardization for two-Dimensional electrophoresis: High-Resolution Two-Dimensional Electrophoretic Mapping of Human proteins
- Author
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Anderson, N L, Edwards, JJ, Giometti, C S, Willard-Gallo, Karen, Tollaksen, SL, Nance, SL, Hickman, B.J., Taylor, J, Coulter, B, Scandora, A E, Anderson, N G, Anderson, N L, Edwards, JJ, Giometti, C S, Willard-Gallo, Karen, Tollaksen, SL, Nance, SL, Hickman, B.J., Taylor, J, Coulter, B, Scandora, A E, and Anderson, N G
- Abstract
info:eu-repo/semantics/published
- Published
- 1980
19. Cytoskeletal proteins from human skin fibroblasts, peripheral blood leukocytes, and a lymphoblastoid cell line compared by two-dimensional gel electrophoresis.
- Author
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Giometti, C S, Willard-Gallo, Karen, Anderson, N L, Giometti, C S, Willard-Gallo, Karen, and Anderson, N L
- Abstract
Differences in proteins between cells grown as suspension cultures and those grown as attached cultures were studied by comparing the proteins of detergent-resistant cytoskeletons prepared from peripheral blood leukocytes and a lymphoblastoid cell line (GM607) (both grown as suspension cultures) and those of human skin fibroblasts (grown as attached cultures) by two-dimensional gel electrophoresis. The major cytoskeletal proteins of the leukocytes were also present in the protein pattern of GM607 cytoskeletons. In contrast, the fibroblast cytoskeletal protein pattern contained four groups of proteins that differed from the patterns of the leukocytes and GM607. Three groups (Cytoskf:8--10, 14--16, and :17--18) showed qualitative differences, and the fourth group (Cytoskf:11 and :13) showed quantitative differences. In addition, surface labeling of GM607 and human fibroblasts with 125I demonstrated that substantial amounts of vimentin and actin are exposed at the surface of the attached fibroblasts, but there is little evidence of similar exposure at the surface of the suspension-grown GM607. Cytoskf:11 and :13 in fibroblast preparations were also labeled with the 125I. These results demonstrate some differences in cytoskeletal protein composition between different types of cells could be related to their ability or lack of ability to grow as attached cells in tissue culture., info:eu-repo/semantics/published
- Published
- 1982
20. Analytical techniques for cell fractions. XXVI. A two-dimentional electrophoretic analysis of basic proteins using phosphatidyl choline/urea solubilization.
- Author
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Willard-Gallo, Karen, Giometti, C S, Anderson, N L, O'Connor, T E, Anderson, N G, Willard-Gallo, Karen, Giometti, C S, Anderson, N L, O'Connor, T E, and Anderson, N G
- Abstract
info:eu-repo/semantics/published
- Published
- 1979
21. Muscle protein analysis. II. Two-dimensional electrophoresis of normal and diseased human skeletal muscle.
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Giometti, C S, primary, Bárány, M, primary, Danon, M J, primary, and Anderson, N G, primary
- Published
- 1980
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22. A two-dimensional electrophoretic analysis of the heat-shock-induced proteins of human cells.
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Anderson, N L, primary, Giometti, C S, primary, Gemmell, M A, primary, Nance, S L, primary, and Anderson, N G, primary
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- 1982
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23. Muscle protein analysis. III. Analysis of solubilized frozen-tissue sections by two-dimensional electrophoresis.
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Giometti, C S, primary and Anderson, N G, primary
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- 1981
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24. Protein changes in activated human platelets.
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Giometti, C S, primary and Anderson, N G, primary
- Published
- 1984
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25. Cytoskeletal proteins from human skin fibroblasts, peripheral blood leukocytes, and a lymphoblastoid cell line compared by two-dimensional gel electrophoresis.
- Author
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Giometti, C S, primary, Willard, K E, primary, and Anderson, N L, primary
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- 1982
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- View/download PDF
26. Muscle protein analysis. I. High-resolution two-dimensional electrophoresis of skeletal muscle proteins for analysis of small biopsy samples.
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Giometti, C S, primary, Anderson, N G, primary, and Anderson, N L, primary
- Published
- 1979
- Full Text
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27. Fast to slow change of myosin in nemaline rod myopathy
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Giometti, C. S., primary and Danon, M. J., additional
- Published
- 1983
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28. Two-dimensional electrophoresis
- Author
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Giometti, C. S., primary and Danon, M. J., additional
- Published
- 1984
- Full Text
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29. Analysis of Proteins from Human Muscle Biopsies by Two-Dimensional Electrophoresis.
- Author
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DANON, M. J., GIOMETTI, C. S., and ANDERSON, N. G.
- Published
- 1982
30. Polyacrylamide gel electrophoretic methods in the separation of structural muscle proteins
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Barany, K., Barany, M., and Giometti, C. S.
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- 1995
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31. Two-dimensional gel electrophoresis mapping of proteins isolated from the hyperthermophile Pyrococcus furiosus
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Giometti, C. S., Tollaksen, S. L., Mukund, S., and Zhou, Z. H.
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- 1995
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32. Methanol toxicity and formate oxidation in NEUT2 mice.
- Author
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Cook RJ, Champion KM, and Giometti CS
- Subjects
- Animals, Formates administration & dosage, Mice, Oxidation-Reduction, Oxidoreductases Acting on CH-NH Group Donors deficiency, Oxidoreductases Acting on CH-NH Group Donors genetics, Formates metabolism, Methanol toxicity, Oxidoreductases Acting on CH-NH Group Donors metabolism
- Abstract
NEUT2 mice are deficient in cytosolic 10-formyltetrahydrofolate dehydrogenase (FDH; EC 1.5.1.6) which catalyzes the oxidation of excess folate-linked one-carbon units in the form of 10-formyltetrahydrofolate to CO(2) and tetrahydrofolate (Champion et al., Proc. Natl. Acad. Sci. USA 91, 11338-11342, 1994). The absence of FDH should impair the oxidation of formate via the folate-dependent pathway and as a consequence render homozygous NEUT2 mice more susceptible to methanol toxicity. Normal (CB6-F1) and NEUT2 heterozygous and homozygous mice had essentially identical LD(50) values for methanol, 6.08, 6.00, and 6.03 g/kg, respectively. Normal mice oxidized low doses of [(14)C]sodium formate (ip 5 mg/kg) to (14)CO(2) at approximately twice the rate of homozygous NEUT2 mice, indicating the presence of another formate-oxidizing system in addition to FDH. Treatment of mice with the catalase inhibitor, 3-aminotriazole (1 g/kg ip) had no effect on the rate of formate oxidation, indicating that at low concentrations formate was not oxidized peroxidatively by catalase. High doses of [(14)C]sodium formate (ip 100 mg/kg) were oxidized to (14)CO(2) at identical rates in normal and NEUT2 homozygous mice. Pretreatment with 3-aminotriazole (1 g/kg ip) in this instance resulted in a 40 and 50% decrease in formate oxidation to CO(2) in both normal and homozygous NEUT2 mice, respectively. These results indicate that mice are able to oxidize formate to CO(2) by at least three different routes: (1) folate-dependent via FDH at low levels of formate; (2) peroxidation by catalase at high levels of formate; and (3) by an unknown route(s) which appears to function at both low and high levels of formate. The implications of these observations are discussed in terms of the current hypotheses concerning methanol and formate toxicity in rodents and primates., (Copyright 2001 Academic Press.)
- Published
- 2001
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33. Structural modifications of Methanococcus jannaschii flagellin proteins revealed by proteome analysis.
- Author
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Giometti CS, Reich CI, Tollaksen SL, Babnigg G, Lim H, Yates JR 3rd, and Olsen GJ
- Subjects
- Amino Acid Sequence, Electrophoresis, Gel, Two-Dimensional, Molecular Sequence Data, Protein Conformation, Sequence Homology, Amino Acid, Archaeal Proteins chemistry, Flagellin chemistry, Methanococcus chemistry, Proteome
- Abstract
Methanococcus jannaschii is an autotrophic archaeon originally isolated from an oceanic thermal vent. The primary metabolic pathway for energy production in this hyperthermophilic microbe is methanogenesis from H2 and CO2. As an autotroph, M. jannaschii requires only CO2 as a carbon source for synthesizing all necessary biomolecules. Changes in the environmental availability of these molecules can be expected to activate regulatory mechanisms manifested as the up and down regulation of specific genes and the concomitant increase and decrease in abundance of the corresponding proteins. In our analysis of the proteome of M. jannaschii, we have observed significant changes in the abundance of a common subset of predominant proteins in response to reduced H2 concentration, limited ammonium availability, and the stage of cell growth (exponential compared with stationary). The masses of tryptic peptides from these proteins match those predicted by M. jannaschii genome open reading frames annotated as flagellin B1 (MJ0891) and flagellin B2 (MJ0892). Multiple proteins with different isoelectric points and molecular weights match each of these proteins, and the abundance of these protein variants changes with growth conditions. These data indicate that structural modifications altering both the isoelectric point and size of the M. jannaschii flagellin B1 and B2 proteins occur in response to growth conditions and growth stage of M. jannaschii and further suggest the regulation of M. jannaschii motility through structural modifications of the building blocks of the flagella.
- Published
- 2001
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34. Identification of membrane proteins in the hyperthermophilic archaeon pyrococcus furiosus using proteomics and prediction programs.
- Author
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Holden JF, Poole Ii FL, Tollaksen SL, Giometti CS, Lim H, Yates Iii JR, and Adams MW
- Abstract
Cell-free extracts from the hyperthermophilic archaeon Pyrococcus furiosus were separated into membrane and cytoplasmic fractions and each was analyzed by 2D-gel electrophoresis. A total of 66 proteins were identified, 32 in the membrane fraction and 34 in the cytoplasmic fraction. Six prediction programs were used to predict the subcellular locations of these proteins. Three were based on signal-peptides (SignalP, TargetP, and SOSUISignal) and three on transmembrane-spanning alpha-helices (TSEG, SOSUI, and PRED-TMR2). A consensus of the six programs predicted that 23 of the 32 proteins (72%) from the membrane fraction should be in the membrane and that all of the proteins from the cytoplasmic fraction should be in the cytoplasm. Two membrane-associated proteins predicted to be cytoplasmic by the programs are also predicted to consist primarily of transmembrane-spanning beta-sheets using porin protein models, suggesting that they are, in fact, membrane components. An ATPase subunit homolog found in the membrane fraction, although predicted to be cytoplasmic, is most likely complexed with other ATPase subunits in the membrane fraction. An additional three proteins predicted to be cytoplasmic but found in the membrane fraction, may be cytoplasmic contaminants. These include a chaperone homolog that may have attached to denatured membrane proteins during cell fractionation. Omitting these three proteins would boost the membrane-protein predictability of the models to near 80%. A consensus prediction using all six programs for all 2242 ORFs in the P. furiosus genome estimates that 24% of the ORF products are found in the membrane. However, this is likely to be a minimum value due to the programs' inability to recognize certain membrane-related proteins, such as subunits associated with membrane complexes and porin-type proteins.
- Published
- 2001
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35. Mouse liver selenium-binding protein decreased in abundance by peroxisome proliferators.
- Author
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Giometti CS, Liang X, Tollaksen SL, Wall DB, Lubman DM, Subbarao V, and Rao MS
- Subjects
- Animals, Cell Division, Electrophoresis, Gel, Two-Dimensional, Liver cytology, Liver metabolism, Male, Mass Spectrometry, Mice, Selenium-Binding Proteins, Carrier Proteins metabolism, Liver drug effects, Peroxisome Proliferators pharmacology
- Abstract
Several studies with two-dimensional gel electrophoresis (2-DE) have shown that the abundance of numerous mouse liver proteins is altered in response to treatment with chemicals known to cause peroxisome proliferation. The peptide masses from tryptic digests of two liver proteins showing dramatic decreases in abundance in response to numerous peroxisome proliferators were used to search sequence databases. The selenium-binding protein 2 (SBP2 formerly 56 kDa acetaminophen-binding protein, AP 56) and selenium-binding protein 1 (SBP1 formerly 56 kDa selenium-binding protein, SP 56) in mouse liver, proteins with a high degree of sequence similarity, were the highest ranked identities obtained. Identity with SBP2 was subsequently confirmed by immunodetection with specific antiserum. Treatment of mice with 0.025% ciprofibrate resulted in the more basic of this pair of proteins being decreased to 30% of control abundance while the acidic protein was decreased to 7% of the control amount. Dexamethasone treatment, in contrast, caused increases of 80% and 20% in the abundance of the acidic and basic forms, respectively. Administration of dexamethasone to mice in combination with ciprofibrate produced expression of the acidic SBP2 at 23% of the control level and the basic SBP2 at 36%, a slightly moderated reduction compared with the decrease that occurred with ciprofibrate alone. These data suggest that peroxisome proliferators such as ciprofibrate cause a decrease in the abundance of the SBP2, which leads to increased cell proliferation, even in the presence of an inhibitor such as dexamethasone. Such a decrease in SBP, thought to serve as cell growth regulation factors, could be central to the nongenotoxic carcinogenicity of the peroxisome proliferators observed in rodents.
- Published
- 2000
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36. A comparison of liver protein changes in mice and hamsters treated with the peroxisome proliferator Wy-14,643.
- Author
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Giometti CS, Tollaksen SL, Liang X, and Cunningham ML
- Subjects
- 3-Hydroxyacyl CoA Dehydrogenases metabolism, Amino Acid Sequence, Animals, Cricetinae, Electrophoresis methods, Electrophoresis, Gel, Two-Dimensional, Enoyl-CoA Hydratase metabolism, Hydrogen-Ion Concentration, Liver ultrastructure, Male, Mesocricetus, Mice, Molecular Sequence Data, Multienzyme Complexes metabolism, Peroxisomal Bifunctional Enzyme, Isomerases, Liver drug effects, Liver metabolism, Peroxisome Proliferators pharmacology, Proteins metabolism, Pyrimidines pharmacology
- Abstract
Interspecies differences in the liver response to Wy-14,643, a potent peroxisome proliferator in rats and mice, have been demonstrated. While both rats and mice show dramatic increases in the number of peroxisomes, the activity of peroxisomal enzymes involved in the beta-oxidation of fatty acids, and heptocyte replication, Syrian hamsters have a more moderate peroxisome proliferation response and no sustained increase in cell replication. Rats and mice, but not hamsters, develop hepatocellular carcinoma after prolonged exposure to Wy-14,643. To further characterize this species difference, two-dimensional gel electrophoresis (2-DE) has been used to compare the effect of 14-day exposure to various dietary concentrations of Wy-14,643 on liver protein expression in male mice and hamsters. Digitized images of the 2-DE protein maps were searched for significant changes. The peroxisome bifunctional enzyme (PBE) enoyl CoA hydratase/3-hydroxyacyl dehydrogenase, which migrates to the same position in mouse and hamster liver protein 2-DE patterns, increased in abundance by more than three times the control level in both mice and hamsters. In addition to the quantitative change in PBE, significant quantitative changes (P < 0.001) were found in 49 mouse liver proteins (47 decreasing and 2 increasing) and in 35 hamster liver proteins (27 decreasing and 8 increasing). There was little overlap in the mouse and hamster proteins showing quantitative changes in response to Wy-14,643, with the exception of PBE and one unidentified liver protein with an approximate molecular weight of 50,000. These results show that although peroxisome proliferation occurs in the livers of both mice and hamsters exposed to Wy-14,643, other species-specific changes in proteins occur that are independent of the peroxisome proliferation response and that could be related to species-specific susceptibility or resistance to liver tumor induction.
- Published
- 1998
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37. Analysis of differential protein expression in normal and neoplastic human breast epithelial cell lines.
- Author
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Williams K, Chubb C, Huberman E, and Giometti CS
- Subjects
- Amino Acid Sequence, Amino Acids, Cells, Cultured, Cytoskeletal Proteins metabolism, Female, Heat-Shock Proteins metabolism, Humans, IMP Dehydrogenase metabolism, Molecular Sequence Data, Sequence Analysis, Subcellular Fractions, Tumor Cells, Cultured, Breast Neoplasms metabolism, Epithelial Cells metabolism, Neoplasm Proteins metabolism
- Abstract
High-resolution two-dimensional gel electrophoresis (2-DE) and database analysis was used to establish protein expression patterns for cultured normal human mammary epithelial cells and thirteen breast cancer cell lines. The Human Breast Epithelial Cell database contains the 2-DE protein patterns, including relative protein abundances, for each cell line, plus a composite pattern that contains all the common and specifically expressed proteins from all the cell lines. Significant differences in protein expression, both qualitative and quantitative, were observed not only between normal cells and tumor cells, but also among the tumor cell lines. Eight percent (56/727) of the consistently detected proteins were found in significantly (P< 0.001) variable levels among the cell lines. Eight proteins present in normal cultured breast epithelial cells were not detected in any of the tumor cell lines. We identified a subset of the differentially expressed proteins using a combination of immunostaining, protein sequencing, comigration, and subcellular fractionation. These identified proteins include the intermediate filament components vimentin and cytokeratins. The cell lines can be classified into four distinct groups based on their intermediate filament protein profile. We also identified heat shock proteins; hsp27 and hsp60 varied in abundance and in some cases in the relative phosphorylation levels among the cell lines. Many of the differentially expressed proteins we identified have roles in cellular proliferation and differentiation, including annexin V, elongation initiation factor 5A, Rho GDP dissociation inhibitor, and prohibitin. We identified inosine-5-monophosphate dehydrogenase in each of the cell lines, and found the levels of this enzyme in the tumor cell lines elevated 2- to 20-fold relative to the levels in normal cells. These results expand the human breast epithelial cell protein database (http:// www.anl.gov/CMB/PMG) which is being built to assist researchers with the identification of abnormal patterns of expression and pathways associated with malignancy.
- Published
- 1998
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38. A two-dimensional electrophoresis database of human breast epithelial cell proteins.
- Author
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Giometti CS, Williams K, and Tollaksen SL
- Subjects
- Breast cytology, Cell Line, Epithelial Cells, Female, Humans, IMP Dehydrogenase analysis, Tetradecanoylphorbol Acetate pharmacology, Tumor Cells, Cultured, Breast chemistry, Computer Communication Networks, Databases, Factual, Electrophoresis, Gel, Two-Dimensional methods, Epithelium chemistry, Neoplasm Proteins analysis, Proteins analysis
- Abstract
As sequencing of the human genome progresses, attention is turning to when and where specific genes are being expressed and how that expression is regulated. The human breast, with the highly specific, but transient, function of milk production (lactation), exemplifies human gene regulation. The molecular mechanisms for the dramatic structural and functional changes involved in shifting from lactation-capable to lactation-incapable tissue are poorly understood, as are the mechanisms that result in deviation from normal breast cell growth into different types of breast neoplasms. We are using quantitative two-dimensional electrophoresis (2-DE) to determine which proteins are present in different types of human breast cells (milk-producing and -nonproducing, estrogen-receptor-positive and -negative, normal and malignant) and which proteins change in abundance in response to stimuli that trigger cell differentiation, growth, or death. A composite map of proteins found in human breast cells is being generated and used as an index of human genes that are differentially expressed, both qualitatively and quantitatively. Proteins found in 15 different types of human breast cells, two from healthy tissue (from milk and reduction mammoplasty tissue) and 13 from tumor tissue, are now included in the composite map. Copies of the human breast epithelial cell protein map are available on the World Wide Web (URL: http:(/)/www.anl.gov/CMB/PMG/ projects/index_hbreast.html) with links to quantitative data and identifications for proteins found to be differentially expressed in these epithelial cells. Links to the Swiss-Prot and enzyme metabolic pathway databases are also provided. The World Wide Web presentation is designed to allow public access to the available 2-DE data together with logical connections to databases providing genome-related information.
- Published
- 1997
- Full Text
- View/download PDF
39. Analysis of proteins from human breast epithelial cells using two-dimensional gel electrophoresis.
- Author
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Giometti CS, Tollaksen SL, Chubb C, Williams C, and Huberman E
- Subjects
- Amino Acid Sequence, Animals, Breast cytology, Cell Differentiation physiology, Epithelial Cells, Epithelium chemistry, Female, Humans, Keratins analysis, Male, Mice, Mice, Nude, Molecular Sequence Data, Neoplasm Transplantation, Sex Characteristics, Tumor Cells, Cultured, Breast chemistry, Electrophoresis, Gel, Two-Dimensional, Neoplasm Proteins analysis, Proteins analysis
- Abstract
The human breast is a highly specialized, complex tissue comprised of a heterogeneous population of cells with varying functions. Interactions between the different cell types, changes in their relative abundance, state of differentiation and function in response to stimuli, as well as the alterations that lead to the aberrant growth associated with malignancy are poorly understood. Two-dimensional gel electrophoresis is being used to compare the proteins found in different breast cells in order to identify the gene products that are common or specific to particular cell types so as to provide markers that will be useful in studies of normal breast cell differentiation and the dedifferentiation or blocked differentiation characteristic of cancer. Protein patterns have been obtained from cells prepared for electrophoresis immediately after isolation from human milk, from cells cultured for fewer than ten passages after isolation from healthy breast tissue removed during reduction mammoplasty, and from cells maintained in long-term tissue culture after isolation from the pleural effusions of patients with breast carcinomas. Differential expression of cytokeratins 8, 18, and 19, shown previously to be predominantly expressed by epithelial cells in the luminal layer of breast tissue, was observed among the cells analyzed. Other non-cytokeratin proteins were also found to be differentially expressed in subsets of both the normal and tumor cells. A composite human breast cell protein pattern was created which includes all the commonly and specifically expressed proteins found in this study. This pattern will be the basis for continuing studies of proteins in the human breast.
- Published
- 1995
- Full Text
- View/download PDF
40. Altered protein expression detected in the F1 offspring of male mice exposed to fission neutrons.
- Author
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Giometti CS, Tollaksen SL, and Grahn D
- Subjects
- Animals, Chi-Square Distribution, Electrophoresis, Gel, Two-Dimensional, Female, Gene Deletion, Liver radiation effects, Male, Mice, Mice, Inbred C57BL, Protein Biosynthesis, Whole-Body Irradiation, Genes radiation effects, Germ-Line Mutation, Liver metabolism, Neutrons, Proteins genetics, Spermatogonia radiation effects
- Abstract
Liver protein expression in F1 offspring arising from spermatogonia exposed to 60 cGy of fission spectrum neutrons from the JANUS reactor was compared to that in offspring from unexposed spermatogonia by using two-dimensional electrophoresis (2DE). Approximately 100 protein spots in 2DE patterns from 167 control offspring and 530 offspring from irradiated sires were monitored for quantitative decreases of 50%, indicative of mutation events causing the loss of one normal copy of a structural gene. Reproducible abnormalities were found only in 3 patterns, all from the offspring of neutron-irradiated sires. Two of the three patterns were from littermates (brother and sister) and both showed an approximately 70% decrease in the amount of liver protein MSN188. The third pattern was from a male mouse sired by a different male and showed an approximately 50% decrease in the abundance of protein MSN94. The decreased abundance of MSN188 and MSN94 was assumed to be due to mutation events referred to as NEUT1 and NEUT2, respectively. Sibling crosses between the 2 mice showing the NEUT1 trait produced offspring with control, decreased and undetectable levels of MSN188 in a ratio of 0.25:0.5:0.25. Test crosses between the F1 offspring expressing the NEUT2 trait back to C57BL/6JANL mice produced offspring expressing normal or decreased amounts of MSN94 in a ratio of 0.5:0.5. Inbreeding of individuals expressing decreased amounts of MSN94 produced mice expressing control, decreased amounts, or no detectable amount of that protein in a ratio of 0.25:0.5:0.25. These results indicate that the decreased abundance of MSN188 or MSN94 originally detected in the F1 offspring is due to a genetically transmissible event. Unlike the heritable protein changes observed previously in the F1 offspring of sires exposed to N-ethyl-N-nitrosourea in which a protein variant was produced, both the NEUT1 and NEUT2 mutation events appear to prevent the production of any protein product. These 2 mutations may thus represent mutation lesions other than point mutations (e.g., deletions or translocations) detectable as quantitative changes in protein expression in the F1 generation.
- Published
- 1994
- Full Text
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41. Mouse liver protein database: a catalog of proteins detected by two-dimensional gel electrophoresis.
- Author
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Giometti CS, Taylor J, and Tollaksen SL
- Subjects
- Animals, Female, Male, Mice, Mice, Inbred Strains, Molecular Weight, Peptide Mapping methods, Sex Characteristics, Species Specificity, Subcellular Fractions chemistry, Databases, Factual, Electrophoresis, Gel, Two-Dimensional, Liver chemistry, Proteins analysis
- Abstract
Alterations in the abundance or structure of mouse liver proteins are being studied using two-dimensional gel electrophoresis (2-DE) to build a database of protein changes correlating with exposure to ionizing radiation or toxic chemicals. Thus far, studies have included the analysis of proteins from the offspring of exposed parents or from the exposed individuals themselves. In order to characterize and identify proteins found altered by such exposures, sex- and strain-related differences in protein patterns have been analyzed, and the subcellular locations of a large portion of the mapped proteins have been determined. As part of these studies, data are collected and stored using a variety of computer hardware and software tools that allow the accumulation of information on the origin of samples, gel identification, experiment description, and protein similarities and differences. This accumulation of information constitutes the mouse liver protein database. Relational database software is used to tie the different facets of the database together so that the results of a variety of experiments can be compared and interrelated. The database optimizes the information obtained from 2-DE gel sets by allowing use of the data for many purposes, including monitoring of gel resolution to ensure the collection of high quality data and correlation of protein effects induced by different agents. This first edition of the Argonne National Laboratory mouse liver protein database lays the foundation for future work and communication that should elucidate the significance of observed protein effects as possible markers of exposure to toxic agents.
- Published
- 1992
- Full Text
- View/download PDF
42. Biochemical characterization of a variant form of cytosolic epoxide hydrolase induced by parental exposure to N-ethyl-N-nitrosourea.
- Author
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Nourooz-Zadeh J, Winder BS, Dietze EC, Giometti CS, Tollaksen SL, and Hammock BD
- Subjects
- Animals, Blotting, Western, Chromatography, Affinity, Clofibrate pharmacology, Electrophoresis, Polyacrylamide Gel, Enzyme Induction, Epoxide Hydrolases chemistry, Epoxide Hydrolases isolation & purification, Female, Isoelectric Point, Mice, Mice, Inbred BALB C, Oxidoreductases metabolism, Pregnancy, Cytosol enzymology, Epoxide Hydrolases metabolism, Ethylnitrosourea toxicity
- Abstract
1. ENU4 mice express a protein variant originally detected in a CBF1 mouse sired by a C57BL/6 mouse exposed to N-ethyl-N-nitrosourea. It appears to be an isoelectric point variant of cytosolic epoxide hydrolase. Affinity purified cytosolic epoxide hydrolase from ENU4 mice has a pI of approximately 5.1 compared to 5.6 in other mouse strains. 2. Clofibrate induced cytosolic epoxide hydrolase to similar levels in five strains of mice. However, CBF1 and ENU4 mice were more sensitive to the induction of palmitoyl CoA oxidase activity. 3. Except for isoelectric point, the physico- and immunochemical properties of cytosolic epoxide hydrolase from ENU4 mice were similar to those of the other mouse strains. Substrate specificities for five of six substrates tested were also similar.
- Published
- 1992
- Full Text
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43. Muscle sarcomere lesions and thrombosis after spaceflight and suspension unloading.
- Author
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Riley DA, Ellis S, Giometti CS, Hoh JF, Ilyina-Kakueva EI, Oganov VS, Slocum GR, Bain JL, and Sedlak FR
- Subjects
- Animals, Atrophy pathology, Edema pathology, Edema physiopathology, Hindlimb physiology, Mitochondria, Muscle ultrastructure, Muscle Contraction physiology, Muscles pathology, Muscles ultrastructure, Myosins metabolism, Organ Size, Rats, Sarcomeres ultrastructure, Staining and Labeling, Muscles physiology, Sarcomeres physiology, Space Flight, Thrombosis physiopathology, Weightlessness adverse effects
- Abstract
Spaceflight (flight) and tail suspension-hindlimb unloading (unloaded) produced significant decreases in fiber cross-sectional areas of the adductor longus (AL), a slow-twitch antigravity muscle. However, the mean wet weight of the flight AL muscles was near normal, whereas that of the suspension unloaded AL muscles was significantly reduced. Interstitial edema within the flight AL, but not in the unloaded AL, appeared to account for this apparent disagreement. In both experimental conditions, the slow-twitch oxidative fibers atrophied more than the fast-twitch oxidative-glycolytic and fast-twitch glycolytic fibers. Immunostaining showed that slow-twitch oxidative fibers expressed fast myosin, producing hybrid fibers containing slow and fast myosin isoforms. Two-dimensional gel electrophoresis of flight AL muscles revealed increased content of fast myosin light chains and decreased amounts of slow myosin light chains and fatty acid-binding protein. In the flight AL, absolute mitochondrial content decreased, but the relatively greater breakdown of myofibrillar proteins maintained mitochondrial concentration near normal in the central intermyofibrillar regions of fibers. Subsarcolemmal mitochondria were preferentially lost and reduced below normal concentration. Elevated fiber immunostaining for ubiquitin conjugates was suggestive of ubiquitin-mediated breakdown of myofibrillar proteins. On return to weight bearing for 8-11 h, the weakened atrophic muscles exhibited eccentric contraction-like lesions (hyperextension of sarcomeres with A-band filaments pulled apart and fragmented), tearing of the supporting connective tissue, and thrombosis of the microcirculation. Segmental necrosis of muscle fibers, denervation of neuromuscular junctions, and extravasation of red blood cells were minimal. Lymphocyte antibody markers did not indicate a significant immune reaction. The flight AL exhibited threefold more eccentric-like lesions than the unloaded AL; the high reentry G forces experienced by the flight animals, but not the unloaded group, possibly accounted for this difference. Muscle atrophy appears to increase the susceptibility to form eccentric contraction-like lesions after reloading; this may reflect weakening of the myofibrils and extracellular matrix. Microcirculation was also compromised by spaceflight, such that there was increased formation of thrombi in the post-capillary venules and capillaries. This blockage led to edema by 8-11 h after resumption of weight bearing by the COSMOS 2044 rats. The present findings indicate that defective microcirculation most likely accounted for the extensive tissue necrosis and microhemorrhages observed for COSMOS 1887 rats killed 2 days after landing.
- Published
- 1992
- Full Text
- View/download PDF
44. Use of principal components analysis for mutation detection with two-dimensional electrophoresis protein separations.
- Author
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Taylor J and Giometti CS
- Subjects
- Animals, Evaluation Studies as Topic, Female, Male, Mice, Multivariate Analysis, Mutation, Electrophoresis, Gel, Two-Dimensional statistics & numerical data, Proteins genetics, Proteins isolation & purification
- Abstract
The application of two-dimensional electrophoresis (2-DE) to mutation detection requires the capability to monitor each protein in a 2-DE pattern for significant changes in abundance indicative of a mutation event. Previously, mutation searches were done using a univariate outlier detection method in which each protein spot was considered independently in a classical outlier search. An alternative approach to analysis of 2-DE patterns for quantitative changes is a multivariate procedure which takes advantage of the observation that protein spots in a 2-DE pattern often represent correlated rather than independent measurements. We have compared the efficiency of univariate and multivariate procedures for mutation detection using data from the Argonne National Laboratory 2-DE database of mouse liver proteins. Analyses involving a total of over 1500 gels were performed to compare the performance of a multivariate method based on principal components analysis (PCA) with the univariate method. Up to 279 spots from each pattern were used for PCA. First, a simulation was performed to assess the detection efficiency of PCA for single protein spots decreased in abundance by 50%. Then, the ability to detect actual mutations was tested using eight confirmed mutations. Results show that, compared to a univariate approach to analysis of data from the mouse model system, the multivariate method increases the number of protein spots on each 2-DE pattern that can be monitored for quantitative changes indicative of mutations by compensating for variables that contribute to the background quantitative variability of protein spots.
- Published
- 1992
- Full Text
- View/download PDF
45. Progesterone-dependent sexual behavior and protein patterns in the ventromedial hypothalamus of the adult female rat.
- Author
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Montemayor ME, Giometti CS, Taylor J, and Roy EJ
- Abstract
Controversy exists concerning mechanisms by which progesterone exerts central nervous system effects on behavior. Progesterone may affect behavior by genomic regulation of protein synthesis. Alternatively, it may work through nongenomic mechanisms, consistent with its short latency to act. In the present study, we have examined the hypothesis that progesterone facilitation of sexual behavior is correlated with modification of the synthesis of specific proteins in the ventromedial hypothalamus (VMH). Ovariectomized rats were treated with either estradiol (4 mug/kg at 0 and 18 h) or estradiol (at 0 and 18 h) plus progesterone (2 mg/kg at 37 h). (35)S-labeled cysteine and methionine were bilaterally infused into the VMH at 37 h (the time of progesterone administration). Following 4 h of infusion, animals were tested for sexual behavior and sacrificed. Newly synthesized VMH proteins were separated by two-dimensional gel electrophoresis followed by fluorography. Analysis of approximately 660 spots/fluorogram in two independent replications indicated that no qualitative or quantitative changes in protein synthesis occurred in response to progesterone. In each replication statistical analysis suggested that the abundance of several proteins may have changed, but no specific proteins were changed in abundance in both replications. Within the range of this technique (10-100 kDa and 4.8-6.7 apparent pI) we found no evidence that progesterone causes alterations in VMH protein synthesis.
- Published
- 1991
- Full Text
- View/download PDF
46. Quantitation of human leukocyte proteins after silver staining: a study with two-dimensional electrophoresis.
- Author
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Giometti CS, Gemmell MA, Tollaksen SL, and Taylor J
- Subjects
- Electrophoresis, Gel, Two-Dimensional methods, Humans, Isoelectric Focusing methods, Molecular Weight, Silver, Staining and Labeling, Tropomyosin blood, Tropomyosin isolation & purification, Blood Platelets chemistry, Blood Proteins analysis, Leukocytes chemistry
- Abstract
The quantitative attributes of human leukocyte proteins detected by silver staining two-dimensional electrophoresis (2-DE) gels were studied by using computer-assisted data analysis. Experiments included (a) analysis of replicate patterns of the same sample, (b) analysis of different dilutions of the same sample, and (c) analysis of samples from different individuals. Over 200 proteins were observed to have coefficients of variation (CV) less than or equal to 15% when data from replicate patterns were analyzed. In contrast, 8 proteins had CV values of less than or equal to 15% when data from different samples were analyzed. The dilution experiment showed that a majority of the proteins detected with some consistency (i.e., observed in at least 80% of the patterns) have a linear relationship between the amount of protein loaded onto a 2-DE gel and the spot volume in the final 2-DE pattern. The slope of the curves and the deviation from linearity were found to be quite protein-specific. These results indicate that optimization of sample purity and minimization of staining protocol variables are required to limit the background quantitative variability between and within 2-DE runs to a level that will allow detection of quantitative changes indicative of biological responses.
- Published
- 1991
- Full Text
- View/download PDF
47. Association of WR-1065 with CHO AA8 cells, nuclei, and nucleoids.
- Author
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Meechan PJ, Vaughan AT, Giometti CS, and Grdina DJ
- Subjects
- Animals, Cell Line, Cells, Cultured metabolism, Cell Nucleus metabolism, Mercaptoethylamines pharmacokinetics, Radiation-Protective Agents pharmacokinetics
- Abstract
The radioprotector WR-1065 (N-(2-mercaptoethyl)-1,3-diaminopropane) has been shown to be the active moiety involved in protecting mammalian cells from the cytotoxic and mutagenic effects of ionizing radiation after administration of WR-1065 or the phosphorylated form, WR-2721. Initial experiments demonstrated that, in our hands, WR-1065 protects Chinese hamster AA8 cells from killing by (a) mechanism(s) other than induction of hypoxia. AA8 cells were then incubated in the presence of [14C]WR-1065 to determine whether association of WR-1065 in vivo was random or targeted to the nucleus or the nuclear matrix. The kinetics of incorporation of labeled material showed rapid incorporation for the first 30 min and little, if any, additional incorporation over the next 2.5 h. Examination of nuclei and nucleoids generated from the AA8 cells indicated that approximately 10% of the drug was localized in the nucleus and the drug that remained was not dislodged with repeated washes of the filters. Association kinetics of the drug with nuclei and nucleoids indicated that there was little increase in drug association with time, suggesting that there may be a limited number of strong association sites in the nucleus, but these sites are either with DNA or with matrix proteins. Exposure of the AA8 cells to 6 Gy of 60Co gamma rays did not significantly alter the association of the drug with AA8 cells. Incubating AA8 cells in [14C]WR-1065 for 30 min and then incubating in drug-free medium indicated that nearly all of the drug was lost from cells within the first 5 min of incubation in drug-free medium. The low level of tightly bound matrix-associated label may be important in generating alterations in matrix organization that have been observed previously in this laboratory.
- Published
- 1991
48. A comparative study of the effects of clofibrate, ciprofibrate, WY-14,643, and di-(2-ethylhexyl)-phthalate on liver protein expression in mice.
- Author
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Giometti CS, Taylor J, Gemmell MA, Tollaksen SL, Lalwani ND, and Reddy JK
- Subjects
- Animals, Clofibric Acid pharmacology, Electrophoresis, Gel, Two-Dimensional, Female, Fibric Acids, Hypolipidemic Agents pharmacology, Liver metabolism, Male, Membrane Proteins biosynthesis, Mice, Microbodies drug effects, Clofibric Acid analogs & derivatives, Diethylhexyl Phthalate pharmacology, Liver drug effects, Membrane Proteins drug effects, Pyrimidines pharmacology
- Abstract
Chemical, physical, or mechanical insults to a cellular system cause changes in the rates of protein synthesis or degradation. To define patterns of quantitative protein changes and identify the biochemical basis of those changes, we are studying fluctuations in mouse liver protein abundance in response to factors such as stages of maturation, changes in diet, and exposure to chemicals. Two-dimensional gel electrophoresis (2DE) together with computer-assisted data analysis is used to collect quantitative data from groups of 2DE patterns. Our analysis of liver protein expression in mice treated with four different chemicals known to cause peroxisome proliferation serves as an example of the changes in protein expression that can be monitored by using this approach. The database we are building indicates that changes specific to a particular treatment as well as generalized effects can be identified, providing the opportunity to identify patterns of protein expression that are indicators of biological response to different types of cellular insults.
- Published
- 1991
49. Quantitative changes in sets of proteins as markers of biological response.
- Author
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Giometti CS, Taylor J, Gemmell MA, Tollaksen SL, Lalwani ND, and Reddy JK
- Subjects
- Animals, Biomarkers, Clofibrate toxicity, Clofibric Acid analogs & derivatives, Clofibric Acid toxicity, Diethylhexyl Phthalate toxicity, Electrophoresis, Gel, Two-Dimensional, Fibric Acids, Liver chemistry, Liver drug effects, Liver Neoplasms, Experimental chemically induced, Male, Mice, Proteins analysis, Pyrimidines toxicity, Carcinogens toxicity, Proteins drug effects
- Published
- 1991
50. Characterization of a protein that appears in the nervous system of the moth Manduca sexta coincident with neuronal death.
- Author
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Montemayor ME, Fahrbach SE, Giometti CS, and Roy EJ
- Subjects
- Aging, Animals, Electrophoresis, Gel, Two-Dimensional, Ganglia growth & development, Molecular Weight, Cell Survival, Moths growth & development, Nerve Tissue Proteins isolation & purification, Nervous System growth & development, Neurons cytology
- Abstract
Two-dimensional gel electrophoresis was used to locate potential neuronal death-related proteins in the moth Manduca sexta. Protein patterns of ganglia of pharate adult moths (taken prior to adult ecdysis) compared with protein patterns of one-day-old adults revealed reproducible changes in protein patterns. An acidic protein of approximately 40,000 Da was present in all samples from adult moths undergoing neuronal death and essentially absent from pharate adult samples.
- Published
- 1990
- Full Text
- View/download PDF
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