Selected Bibliography

VIRUS ISOLATION AND CHARACTERIZATION

1. Centrifugal systems in blood fractionation. Problems and prospects. Anderson, N. G. Vox Sang 23(1), 135-40. (1972).

2. Mass purification of nucleopolyhedrosis virus inclusion bodies in the K-series centrifuge. Breillatt, J. P., Brantley, J. N., Mazzone, H. M., Martignoni, M. E., Franklin, J. E., and Anderson, N. G. Appl Microbiol 23(5), 923-30. (1972).

3. Separation of Treponema pallidum from tissue substances by continuous-flow zonal centrifugation. Thomas, M. L., Clark, J. W. Jr., Cline, G. B., Anderson, N. G., and Russell, H. Appl Microbiol 23(4), 714-20. (1972).

4. Isopycnometric serology: a new technique based on buoyant density changes in latex beads. Anderson, N. G. and Breillatt, J. P. Nat New Biol 231(21), 112-4. (1971).

5. Evolutionary significance of virus infection. Anderson, N. G. Nature 227(265), 1346-7. (1970).

6. Purification of influenza virus in the K-II zonal centrifuge. Gerin, J. L. and Anderson, N. G. Nature 221(187), 1255-6. (1969).

7. Isolation of T antigen from solid hamster tumors induced by adenovirus type 31. Jainchill, J. L., Candler, E. L., and Anderson, N. G. Proc Soc Exp Biol Med 130(3), 770-5. (1969).

8. Detection of persistent infections using biophysical methods. Anderson, N. G. Natl Cancer Inst Monogr 29, 393-403. (1968).

9. Evaluation of "virus-like" particles in the plasmas of 255 patients with leukemia and related diseases. Newell, G. R., Harris, W. W., Bowman, K. O., Boone, C. W., and Anderson, N. G. N Engl J Med 278(22), 1185-91. (1968).

10. Purification of large quantities of influenza virus by density gradient centrifugation. Reimer, C. B., Baker, R. S., Van Frank, R. M., Newlin, T. E., Cline, G. B. , and Anderson, N. G. J Virol 1(6), 1207-16. (1967).

11. Respiratory syncytial virus isolation by combined continuous flow-isopycnic banding centrifugation. Cline, G. B., Coates, H., Anderson, N. G., Chanock, R. M., and Harris, W. W. J Virol 1(4), 659-64. (1967).

12. Unusual particles in human plasma from leukemia and lymphosarcoma. Harris, W. W., Anderson, N. G., Bartlett, T. W., Rutenberg, E. L., McCauley, L. L., and Kniseley, R. M. Natl Cancer Inst Monogr 21, 389-94. (1966).

13. Problems in biocontainment. Cho, N., Barringer, H. P., Amburgey, J. W., Cline, G. B., Anderson, N. G., McCauley, L. L., Stevens, R. H., and Swartout, W. M. Natl Cancer Inst Monogr 21, 485-502. (1966).

CENTRIFUGE TECHNOLOGY DEVELOPMENT

1. Anderson, Norman G. The development of zonal centrifuges and ancillary systems for tissue fractionation and analysis. (1966). Washington], U.S. Dept. of Health, Education, and Welfare, Public Health Service, National Cancer Institute; [for sale by the Superintendent of Documents.

2. K-series centrifuges. IV. Measurement and control of temperature. Brantley, J. N., Willis, D. D., Breillatt, J. P., Gibson, R. F., Patrick, L. C., and Anderson, N. G. Anal Biochem 36(2), 434-42. (1970).

3. K-series centrifuges. 3. Effect of core taper on particle capture efficiency. Perardi, T. E. and Anderson, N. G. Anal Biochem 34, 112-22. (1970).

4. K-series centrifuges. II. Performance of the K-II rotor. Perardi, T. E., Leffler, R. A., and Anderson, N. G. Anal Biochem 32(3), 495-511. (1969).

5. K-series centrifuges. I. Development of the K-II continuous-sample-flow-with-banding centrifuge system for vaccine purification. Anderson, N. G., Waters, D. A., Nunley, C. E., Gibson, R. F., Schilling, R. M., Denny, E. C., Cline, G. B., Babelay, E. F., and Perardi, T. E. Anal Biochem 32(3), 460-94. (1969).

6. Analytical techniques for cell fractions. XV. Rotor B-XXIX--a new high-resolution zonal centrifuge rotor for virus isolation and cell fractionation. Anderson, N. G., Nunley, C. E., and Rankin, C. T. Jr. Anal Biochem 31(1), 255-71. (1969).

7. Use of the zonal centrifuge to separate particles containing transplantation antigen. Popp, R. A., Popp, D. M., Anderson, N. G., and Elrod, L. H. Biochim Biophys Acta 184(3), 625-33. (1969).

8. Analytical techniques for cell fractions. 13. Rotor A-XVI, a plastic gradient-reorienting rotor for isolating nuclei. Elrod, L. H., Patrick, L. C., and Anderson, N. G. Anal Biochem 30(2), 230-48. (1969).

9. Transport phenomena in zonal centrifuge rotors. II. A mathematical analysis of the areas of isodensity surfaces in reorienting gradient systems. Hsu, H. W. and Anderson, N. G. Biophys J 9(2), 173-88. (1969).

10. Flow studies in the K-II centrifuge. ORNL-4419. Perardi, T. E. and Anderson, N. G. ORNL-NSIC Rep , 33-4. (1969).

11. Analytical techniques for cell fractions. XI. Rotor B-23--a zonal centrifuge rotor for center or edge unloading. Anderson, N. G., Rankin, C. T. Jr., Brown, D. H., Nunley, C. E., and Hsu, H. W. Anal Biochem 26(3), 415-9. (1968).

12. Isolation of a membrane fraction enriched in nerve-end membranes from rat brain by zonal cetrifugation. Cotman, C., Mahler, H. R., and Anderson, N. G. Biochim Biophys Acta 163(2), 272-5. (1968).

13. Analytical techniques for cell fractions. 8. Analytical differential centrifugation in angle-head rotors. Anderson, N. G. Anal Biochem 23(1), 72-83. (1968).

14. The molecular anatomy of cells and tissues. (The MAN program). ORNL-4171 Spec. Anderson, N. G. Ornl , 1-133. (1968).

15. Isolation of rat liver cell membranes by combined rate and isopycnic zonal centrifugation. Anderson, N. G., Lansing, A. I., Lieberman, I., Rankin, C. T., and Elrod, H. Wistar Inst Symp Monogr 8, 23-35. (1968).

16. Analytical techniques for cell fractions. V. Characteristics of the B-XIV and B-XV zonal centrifuge rotors. Anderson, N. G., Waters, D. A., Fisher, W. D., Cline, G. B., Nunley, C. E., Elrod, L. H., and Rankin, C. T. Jr. Anal Biochem 21(2), 235-52. (1967).

17. Analytical techniques for cell fractions. VII. A simple gradient-forming apparatus. Anderson, N. G. and Rutenberg, E. Anal Biochem 21(2), 259-65. (1967).

18. Analytical techniques for cell fractions. VI. Multiple gradient-distributing rotor (B-XXI). Candler, E. L., Nunley, C. E., and Anderson, N. G. Anal Biochem 21(2 ), 253-8. (1967).

19. Preparative zonal centrifugation. Anderson, N. G. Methods Biochem Anal 15, 271-310. (1967).

20. Improved continuous flow centrifugation with banding. Cline, G. B., Nunley, C. E., and Anderson, N. G. Nature 212(61), 487-9. (1966).

21. Zonal centrifuges and other separation systems. Anderson, N. G. Science 154(745 ), 103-12. (1966).

22. Separation of subcellular components and viruses by combined rate- and isopycnic-zonal centrifugation. Anderson, N. G., Harris, W. W., Barber, A. A., Rankin, C. T. Jr., and Candler, E. L. Natl Cancer Inst Monogr 21, 253-83. (1966).

23. Lipid peroxidation in rat tissue particulates separated by zonal centrifugation. Barber, A. A., Rankin, C. T. Jr., and Anderson, N. G. Natl Cancer Inst Monogr 21, 333-44. (1966).

24. Isolation of native glycogen by combined rate-zonal and isopycnic centrifugation. Barber, A. A., Harris, W. W., and Anderson, N. G. Natl Cancer Inst Monogr 21, 285-302. (1966).

25. Continuous-flow centrifugation combined with isopycnic banding: rotors B-8 and B-IX. Anderson, N. G., Barringer, H. P., Amburgey, J. W. Jr., Cline, G. B., Nunley, C. E., and Berman, A. S. Natl Cancer Inst Monogr 21, 199-216. (1966).

26. An introduction to particle separations in zonal centrifuges. Anderson, N. G. Natl Cancer Inst Monogr 21, 9-39. (1966 ).

27. Design of the B-V continuous-flow centrifuge system. Barringer, H. P., Anderson, N. G., and Nunley, C. E. Natl Cancer Inst Monogr 21, 191-8. (1966).

28. Zonal rotors with removable seals: rotors B-X and B-XI. Barringer, H. P., Anderson, N. G., Nunley, C. E., Ziehlke, K. T., and Dritt, W. S. Natl Cancer Inst Monogr 21, 165-74. (1966).

29. The design and operation of the B-IV zonal centrifuge system. Anderson, N. G., Barringer, H. P., Babelay, E. F., Nunley, C. E., Bartkus, M. J., Fisher, W. D., and Rankin, C. T. Jr. Natl Cancer Inst Monogr 21, 137-64. (1966).

30. The development of low-speed "A" series zonal rotors. Anderson, N. G., Barringer, H. P., Cho, N., Nunley, Ce Babelay Ef, Canning, R. E., and Rankin, C. T. Jr. Natl Cancer Inst Monogr 21, 113-36. (1966).

31. An evaluation of the B-V (continuous-flow) and B-IV (density gradient) rotors by use of live polio virus. Reimer, C. B., Newlin, T. E., Havens, M. L., Baker, R. S., Anderson, N. G., Cline, G. B., Barringer, H. P., and Nunley, C. E. Natl Cancer Inst Monogr 21, 375-88. (1966).

CENTRIFUGAL CLINICAL ANALYZERS

1. Fast analyzers for biochemical analysis. Tiffany, T. O., Burtis, C. A., and Anderson, N. G. Methods Enzymol 31(Pt A), 790-833. (1974).

2. Increased rate of analysis by use of a 42-cuvet GeMASAEC fast analyzer. Burtis, C. A., Johnson, W. F., Attrill, J. E., Scott, C. D., Cho, N., and Anderson, N. G. Clin Chem 17(8), 686-95. (1971).

3. Analytical techniques for cell fractions. XVII. The G-IIC fast analyzer system. Mashburn, D. N., Stevens, R. H., Willis, D. D., Elrod, L. H., and Anderson, N. G. Anal Biochem 35(1), 98-112. (1970).

4. Basic principles of fast analyzers. Anderson, N. G. Am J Clin Pathol 53(5), 778-85. (1970).

5. GeMSAEC: a new analytic tool for clinical chemistry total serum protein with the biuret reaction. Hatcher, D. W. and Anderson, N. G. Am J Clin Pathol 52(6), 645-51. (1969).

6. Computer interfaced fast analyzers. Anderson, N. G. Science 166(903), 317-24. (1969).

7. Analytical techniques for cell fractions. XIV. Use of drainage syphons in a fast-analyzer cuvet-rotor. Anderson, N. G. Anal Biochem 32(1), 59-69. (1969).

8. Analytical techniques for cell fractions. XVI. Preparation of protein-free supernatants with a "Z"-path rotor. Anderson, N. G. Anal Biochem 31(1), 272-8. (1969).

9. The development of automated systems for clinical and research use. Anderson, N. G. Clin Chim Acta 25( 2), 321-30. (1969).

10. Analytical techniques for cell fractions. XII. A multiple-cuvet rotor for a new microanalytical system. Anderson, N. G. Anal Biochem 28(1), 545-62. (1969).

11. Adaptation of GeMSAEC fast analyzers to immunochemical studies. ORNL-4419. Anderson, N. G. ORNL-NSIC Rep , 48-9. (1969).

12. Detection of persistent infections using biophysical methods. Anderson, N. G. Natl Cancer Inst Monogr 29, 393-403. (1968).

13. Purification of large quantities of influenza virus by density gradient centrifugation. Reimer, C. B., Baker, R. S., Van Frank, R. M., Newlin, T. E., Cline, G. B. , and Anderson, N. G. J Virol 1(6), 1207-16. (1967).

14. Respiratory syncytial virus isolation by combined continuous flow-isopycnic banding centrifugation. Cline, G. B., Coates, H., Anderson, N. G., Chanock, R. M., and Harris, W. W. J Virol 1(4), 659-64. (1967).

15. Unusual particles in human plasma from leukemia and lymphosarcoma. Harris, W. W., Anderson, N. G., Bartlett, T. W., Rutenberg, E. L., McCauley, L. L., and Kniseley, R. M. Natl Cancer Inst Monogr 21, 389-94. (1966).

PROTEOMICS

1. Back to the future: the human protein index (HPI) and the agenda for post-proteomic biology. Anderson, N. G., Matheson, A., and Anderson, N. L. Proteomics 1(1), 3-12. (2001).

The effort to produce an index of all human proteins (the human protein index, or HPI) began twenty years ago, before the initiation of the human genome program. Because DNA sequencing technology is inherently simpler and more scalable than protein analytical technology, and because the finiteness of genomes invited a spirit of rapid conquest, the notion of genome sequencing has displaced that of protein databases in the minds of most molecular biologists for the last decade. However, now that the human genome sequence is nearing completion, a major realignment is under way that brings proteins back to the center of biological thinking. Using an influx of new and improved protein technologies--from mass spectrometry to re-engineered two-dimensional (2-D) gel systems, the original objectives of the HPI have been expanded and the time frame for its execution radically shortened. Several additional large scale technology efforts flowing from the HPI are also described.

2. Analytical techniques for cell fractions. XXIII. A stable thermal gradient device for heat denaturation studies on proteins. Anderson, N. L., Eisler, W. J., and Anderson, N. G. Anal Biochem 91(2), 441-5. (1978).

The ISO-DALT two-dimensional electrophoretic system (1,2), based on the method of O'Farrell (3), is capable of performing large numbers of analysis on complex mixtures of proteins. However, both separations employed are carried out under dissociating or denaturing conditions and no enzyme activities are readily observable in the analyzed proteins. In order to identify the spots corresponding to particular enzymes, it is therefore necessary to employ some nondestructive resolving technique first and as a second step to perform both enzyme and two-dimensional electrophoretic analyses on the fractions generated. By correlating enzyme activity with intensity of various spots on the two-dimensional gels throughout the series of initial fractions, identifications, can be made. This approach, unlike the more direct immunoprecipitation methods (4), requires the running of large numbers of enzyme analyses and two-dimensional gels and some convenient initial resolving procedure. Convenient and rapid techniques for the analyses (5,6) and gels (1,2) have been described previously in this series and elsewhere. This paper deals with the use of selective denaturation in a temperature gradient as an initial resolving procedure and describes a simple thermal gradient device for generating such a gradient.

3. Proteome and proteomics: new technologies, new concepts, and new words. Anderson, N. L. and Anderson, N. G. Electrophoresis 19(11), 1853-61. (1998).

The goal of proteomics is a comprehensive, quantitative description of protein expression and its changes under the influence of biological perturbations such as disease or drug treatment. Quantitative analysis of protein expression data obtained by high-throughput methods has led us to define the concept of "regulatory homology" and use it to begin to elucidate the basic structure of gene expression control in vivo. Such investigations lay the groundwork for construction of comprehensive databases of mechanisms (cataloguing possible biological outcomes), the next logical step after the soon to be completed cataloguing of genes and gene products. Mechanism databases provide a roadmap towards effective therapeutic intervention that is more direct than that offered by conventional genomics approaches.

4. Twenty years of two-dimensional electrophoresis: past, present and future. Anderson, N. G. and Anderson, N. L. Electrophoresis 17(3), 443-53. (1996).

5. Simultaneous measurement of hundreds of liver proteins: application in assessment of liver function. Anderson, N. L., Taylor, J., Hofmann, J. P., Esquer-Blasco, R., Swift, S., and Anderson, N. G. Toxicol Pathol 24(1), 72-6. (1996).

Proteins implement most biological functions at the molecular level. As one might expect based on this fact, it appears that the altered functional states associated with toxic effects involve changes in the abundance or structure of proteins. Although numerous specific assays exist to measure changes in the abundance of individual proteins, practical limitations have prevented widespread use of multiple protein assays for the global characterization of toxicity. Recent developments in protein analytical technology are rapidly changing this picture. Two-dimensional gel electrophoresis, a technique capable of resolving and quantitating hundreds of proteins simultaneously, is becoming an automated, high-throughput tool. In parallel, techniques have been developed that allow the resulting deluge of protein measurements to be organized into a prototype Molecular Effects Database describing xenobiotic effects in rodent liver. This database can detect, classify, and characterize a broad range of liver toxicity mechanisms. It currently contains approximately 10 million protein measurements, including data on the liver effects of 43 compounds, with a further 50 compounds to be added in 1995. Observed effects range from very broad (sex steroids alter levels of 45% of all liver proteins) to very specific (e.g., hepatic hydroxymethyl glutaryl coenzyme A reductase inhibitors). Companion 2-dimensional databases describing rodent brain and kidney have been initiated, as have linkages to the genomic sequence databases. Assimilation of this approach into research and regulatory toxicology poses an interesting challenge--one that is likely to lead to a radically more sophisticated understanding of toxicity and its biological basis.

6. An updated two-dimensional gel database of rat liver proteins useful in gene regulation and drug effect studies. Anderson, N. L., Esquer-Blasco, R., Hofmann, J. P., Meheus, L., Raymackers, J., Steiner, S. , Witzmann, F., and Anderson, N. G. Electrophoresis 16(10), 1977-81. (1995).

We have improved upon the reference two-dimensional (2-D) electrophoretic map of rat liver proteins originally published in 1991 (N. L. Anderson et al., Electrophoresis 1991, 12, 907-930). A total of 53 proteins (102 spots) are now identified, many by microsequencing. In most cases, spots cut from wet, Coomassie Blue stained 2-D gels were submitted to internal tryptic digestion [2], and individual peptides, separated by high-performance liquid chromatography (HPLC), were sequenced using a Perkin-Elmer 477A sequenator. Additional spots were identified using specific antibodies.

7. A two-dimensional gel database of human plasma proteins. Anderson, N. L. and Anderson, N. G. Electrophoresis 12(11), 883-906. (1991).

An updated two-dimensional electrophoretic map of human plasma proteins is presented, together with a complete listing of the individual protein spots, their locations, size and isoelectric points relative to internal charge standards. Forty-nine polypeptide species are identified, many consisting of multiple spots differing in glycosylation or sequence (e.g., immunoglobulins). A further series of 35 as yet uncharacterized proteins is indicated.

8. A two-dimensional gel database of rat liver proteins useful in gene regulation and drug effects studies. Anderson, N. L., Esquer-Blasco, R., Hofmann, J. P., and Anderson, N. G. Electrophoresis 12(11), 907-30. (1991).

A standard two-dimensional (2-D) protein map of Fischer 344 rat liver (F344MST3) is presented, with a tabular listing of more than 1200 protein species. Sodium dodecyl sulfate (SDS) molecular mass and isoelectric point have been established, based on positions of numerous internal standards. This map has been used to connect and compare hundreds of 2-D gels of rat liver samples from a variety of studies, and forms the nucleus of an expanding database describing rat liver proteins and their regulation by various drugs and toxic agents. An example of such a study, involving regulation of cholesterol synthesis by cholesterol-lowering drugs and a high-cholesterol diet, is presented. Since the map has been obtained with a widely used and highly reproducible 2-D gel system (the Iso-Dalt system), it can be directly related to an expanding body of work in other laboratories.

9. Effects of toxic agents at the protein level: quantitative measurement of 213 mouse liver proteins following xenobiotic treatment. Anderson, N. L., Giere, F. A., Nance, S. L., Gemmell, M. A., Tollaksen, S. L., and Anderson, N. G. Fundam Appl Toxicol 8(1), 39-50. (1987).

By analyzing two-dimensional electrophoretic patterns of mouse liver proteins with a computerized image analysis system, we have observed quantitative changes in the abundance of more than 70 proteins in mice treated with various agents. Aroclor 1254, a mixture of polychlorinated biphenyls known to induce a broad spectrum of microsomal activity, induces the largest group of changes (60 proteins altered at p less than 0.001 significance). Phenobarbital produces a small set of characteristic changes that forms part of the much larger Aroclor 1254 effect. Ibuprofen treatment produces a phenobarbital-like pattern of change, with the addition of at least one protein change not observed with any of the other treatments. Cycloheximide and carbon tetrachloride each induces a different characteristic pattern of protein alteration. We have assigned most of the mouse liver proteins to a specific subcellular fraction, and it appears that the predominant class of proteins altered by each compound is present in the soluble phase, rather than in the microsomal fraction. The ability to survey large numbers of tissue proteins for involvement in pharmacologic and toxic effects may allow a more comprehensive understanding of the mechanisms of action in vivo and provide new markers of tissue damage.

10. "Map" of proteins resolved from human chorionic villi by two-dimensional electrophoresis. Trnka, P. L., Pergament, E., and Anderson, N. G. Clin Chem 30(12 Pt 1), 2040-2. (1984).

Two-dimensional electrophoresis was applied to specimens of human chorionic villi obtained during the first trimester of gestation, the object being to simultaneously map several hundred polypeptide gene products. Genetically normal specimens were homogenized in a urea-based denaturant and the supernates were electrophoresed with use of the "ISO-DALT" system. Four categories of proteins are distinguished on the map: previously identified proteins present in chorionic villi and other cell types; unidentified proteins present in chorionic villi and other cell types; proteins present in chorionic villi and amniotic fluid but not in other cell types; and proteins probably originating from the amnio-chorionic plate. The reference map for chorionic villi provided in this study may serve as the basis of determining whether genetic analyses conducted in the first trimester accurately represent the fetal genotype.

11. Protein changes in activated human platelets. Giometti, C. S. and Anderson, N. G. Clin Chem 30(12 Pt 1), 2078-83. (1984).

Using two-dimensional electrophoresis, we mapped both the total and the cytoskeletal proteins of human platelets before and after activation with thrombin or the calcium ionophore A23187. Activation resulted in increased abundance of the phosphorylated form of myosin light chains with an approximate molecular mass of 20 kDa, decreased abundance of two proteins with molecular masses of approximately 18 and 25 kDa, and, in the case of activation with thrombin, the appearance of a new chain of protein spots (named "Thromb:1"). The latter, found associated with isolated detergent-insoluble cytoskeletons, reacted with antibody to human fibrinogen and thus were identified as gamma-gamma dimers of fibrin. The total number of proteins associated with the cytoskeleton increased after activation with either thrombin or A23187, but we observed some differences in which proteins were bound, and for how long.

12. Human muscle proteins: analysis by two-dimensional electrophoresis. Giometti, C. S., Danon, M. J., and Anderson, N. G. Neurology 33(9), 1152-6. (1983).

Proteins from single frozen sections of human muscle were separated by two-dimensional gel electrophoresis and detected by fluorography or Coomassie Blue staining. The major proteins were identical in different normal muscles obtained from either sex at different ages, and in Duchenne and myotonic dystrophy samples. Congenital myopathy, denervation atrophy, polymyositis, and Becker's muscular dystrophy samples, however, showed abnormal myosin light chain compositions, some with a decrease of fast-fiber myosin light chains and others with a decrease of slow-fiber light chains. These protein alterations did not correlate with any specific disease, and may be caused by generalized muscle-fiber damage.

13. Analysis of human leukemic cells by use of high-resolution two-dimensional electrophoresis. I: results of a pilot study. Anderson, N. L., Wiltsie, J. C., Li, C. Y., Willard-Gallo, K. E., Tracy, R. P., Young, D. S., Powers, M. T., and Anderson, N. G. Clin Chem 29(5), 762-7. (1983).

We analyzed mononuclear leukocytes from patients with various human leukemias by high-resolution two-dimensional electrophoresis. Tumor cells of the granulocytic, monocytic, and lymphoid lineages [obtained from chronic granulocytic leukemia in blast transformation, acute monocytic leukemia, and chronic lymphocytic leukemia (CLL), respectively] can be easily recognized by using a series of cell-type marker proteins identified by comparison of fractionated normal cell populations. B and T cell types of CLL could be distinguished, the results correlating well with those obtained by use of monoclonal-antibody staining methods. In two cases representing almost pure B-cells (classical CLL; 0% T, 85% B) and T-cells (cutaneous T-cell leukemia; 77% T, 0% B), 27 of 29 marker proteins showed quantitative B/T differences comparable to those observed in comparisons of normal B-and T-lymphocytes prepared by cell sorting. These results indicate that cells from relatively well-differentiated leukemias show complex patterns of gene expression very similar to those of the corresponding normal cells and strongly support the use of large marker panels in cell-type determination. Less-well-differentiated acute leukemias [such as acute undifferentiated and acute granulocytic (FAB:M1)] appear to yield protein patterns corresponding less closely to recognizable mature cell types, and may show expression of novel proteins related to the state of differentiation.

14. High-resolution protein separation and identification methods applicable to virology. Anderson, N. G. Curr Top Microbiol Immunol 104, 197-217. (1983).

15. A two-dimensional electrophoretic analysis of the heat-shock-induced proteins of human cells. Anderson, N. L., Giometti, C. S., Gemmell, M. A., Nance, S. L., and Anderson, N. G. Clin Chem 28(4 Pt 2), 1084-92. (1982).

Using two-dimensional electrophoresis, we have investigated the responses of human cells in culture to heat shock and to various chemical agents producing a similar effect. These treatments result in the induction of increased synthesis of several specific proteins. One (HShock:1, SDS-molecular mass about 65000) is increased by about 350-fold over the amount in untreated cells. Computer analysis of time-course studies indicates, however, that rates of synthesis of various proteins other than the classical "heat shock proteins" are affected, some of these alterations following time courses quite different from the main (HShock) inductions. The heat shock effect is thus much more complicated than previously realized. We purified the HShock:1 protein from heat-shocked human lymphoblastoid cells, and prepared a rabbit antiserum specific for HShock:1 on nitrocellulose two-dimensional gel transfers of total lymphoblastoid cell protein. A survey of mouse tissues shows high concentrations of an HShock:1-like protein in the testis, and human testes also appears to contain substantial (though lower) concentrations. These results are consistent with the hypothesis (derived from the tissue-culture studies) that the heat shock effect is a general response to the need for increased protein catabolism within the cell. Increased concentrations of HShock:1 are also observed in preparations of blood leukocytes collected from patients after surgery, indicating that some types of physiological trauma may induce the heat shock proteins in man. Using the antiHShock:1 antibody in an immunoassay, it will be possible to systematically examine HShock:1 concentrations in plasma and leukocytes, thereby opening up the possibility of a clinical test based for the first time upon an inducible aspect of cellular gene expression.

16. The Human Protein Index. Anderson, N. G. and Anderson, L. Clin Chem 28(4 Pt 2), 739-48. (1982).

17. Design and implementation of a prototype Human Protein Index. Taylor, J., Anderson, N. L., Scandora, A. E. Jr., Willard, K. E., and Anderson, N. G. Clin Chem 28(4 Pt 2), 861-6. (1982).

This paper describes information-handling aspects of the TYCHO I analysis system (Clin, Chem. 27: 1807--1820, 1981), which analyzes two-dimensional electrophoresis gels, matches the individual protein spots with those in a reference pattern, and stores various information--including spot measurements, identifications, treatment profiles, set memberships, and comments--in a computerized database. This and additional information such as amino acid composition and cellular localization is then accessible from an interactive program that includes a pictorial user interface and presents much of the data in graphical form. Use of the TYCHO I system is illustrated by examples drawn from analyses of gel patterns from human leukocytes.

18. Proteins of human urine. III. Identification and two-dimensional electrophoretic map positions of some major urinary proteins. Edwards, J. J., Tollaksen, S. L., and Anderson, N. G. Clin Chem 28(4 Pt 2), 941-8. (1982).

We mapped the proteins of human urine by high-resolution two-dimensional electrophoresis, utilizing the ISO-DALT system. Wide-range pH gradients and narrow-range acid gradients were both used in the first-dimension separations. The patterns revealed proteins ranging in relative molecular mass from 10 000 to 90 000. Proteins identified in the map included transferrin, albumin, hemopexin, alpha 2-HS glycoprotein, alpha 1-antitrypsin. Gc globulin, alpha 1-acid glycoprotein, Zn alpha 2-glycoprotein, retinol binding protein, beta 2-microglobulin, the immunoglobulin light chains, and MAUP (most acid urinary protein). The use and utility of internal-charge and molecular-mass standards are described. We used electrophoretic transfer of proteins to nitrocellulose sheets and subsequent detection by immunological methods to identify some proteins.

19. Proteins of human milk. I. Identification of major components. Anderson, N. G., Powers, M. T., and Tollaksen, S. L. Clin Chem 28(4 Pt 2), 1045-55. (1982).

Traditionally, human milk proteins are identified largely by reference to bovine milk. Hence, to identify the major proteins in human milk, we subjected human and bovine milk, in parallel, to high-resolution two-dimensional electrophoresis. Isoelectric precipitation at pH 4.6 was our criterion for distinguishing whey proteins from those of the casein complex. The alpha- and beta-caseins were identified on the basis of relative abundance, relative molecular mass, and relative isoelectric points. Kappa casein was identified as a series of four spots, which disappear from bovine skim milk treated with rennin (chymosin; EC 3.4.23.4) during the clotting process. Para kappa-casein does not appear on the standard ISO-DALT pattern after treatment of bovine milk with rennin, but does appear in BASO-DALT pattern, indicating its high isoelectric point. No protein disappeared from ISO-DALT patterns of human milk after rennin treatment, and no new protein comparable to bovine para kappa-casein appeared in the BASO-DALT patterns; this suggests that kappa-casein is absent from human milk. The proteins identified in human milk patterns include the alpha and beta casein families, lactalbumin, albumin, transferrin, IgA, and lactoferrin. Numerous additional proteins seen in patterns for human milk remain to be identified.

20. Proteins of human urine. II. Identification by two-dimensional electrophoresis of a new candidate marker for prostatic cancer. Edwards, J. J., Anderson, N. G., Tollaksen, S. L., von Eschenbach, A. C., and Guevara, J. Jr. Clin Chem 28(1), 160-3. (1982).

A protein series common to the urine and prostatic tissue of 16 of 17 patients with prostatic adenocarcinoma has been identified by high-resolution two-dimensional gel electrophoresis. These proteins, designated PCA-1, have a relative molecular mass in sodium dodecyl sulfate of about 40000. Analyses of urines from eight age-matched controls, seven patients with other ty pes of urogenital malignancies, two patients with benign prostatic hyperplasia, and five patients with malignancies not associated with the urogenital system failed to show PCA-1 in the patterns. These preliminary findings suggest that this protein should be systematically investigated as a candidate marker for prostatic adenocarcinoma in man.

21. Photo/essay. The human protein index. Anderson, N. G. and Anderson, N. L. Jama 246(22), 2620-1 . (1981).

22. Muscle protein analysis. III. Analysis of solubilized frozen-tissue sections by two-dimensional electrophoresis. Giometti, C. S. and Anderson, N. G. Clin Chem 27(11), 1918-21. (1981).

Proteins from frozen histological sections of human muscle were analyzed by two-dimensional gel electrophoresis. Patterns so obtained were identical to those from whole homogenates of muscle prepared from frozen tissue powders that had much higher protein concentrations. To increase the number of proteins visible on gels of samples low in protein content, the gels were silver stained, or the proteins were labeled with [14C]iodoacetamide before electrophoresis and the gels were fluorographed. The latter method allow use of a single frozen-tissue section for two-dimensional electrophoretic analysis and brings the technique closer to practicable clinical use.

23. The TYCHO system for computer analysis of two-dimensional gel electrophoresis patterns. Anderson, N. L., Taylor, J., Scandora, A. E., Coulter, B. P., and Anderson, N. G. Clin Chem 27(11), 1807-20. (1981).

We describe here a computer system for the analysis of high-resolution two-dimensional gel-electrophoresis patterns, with some initial applications. The system (called TYCHO) comprises programs for image acquisition, background subtraction and smoothing, spot detection, gaussian spot modeling, and pattern matching and comparison. It is based on a conventional minicomputer, but makes extensive use of a high-speed array processor in the image-processing and -modeling steps. Used in concert with the ISO-DALT two-dimensional electrophoresis system (Anal. Biochem. 85:331-354, 1978), TYCHO allows quantitative measurement of hundreds of proteins in complex biological samples, and constitutes the initial data-reduction system required for work towards a Human Protein Index.

24. Proteins of human semen. I. Two-dimensional mapping of human seminal fluid. Edwards, J. J., Tollaksen, S. L., and Anderson, N. G. Clin Chem 27(8), 1335-40. (1981).

The proteins in human seminal plasma were mapped by high-resolution two-dimensional electrophoresis (ISO-DALT and BASO-DALT systems). When analyzed under dissociating conditions, samples from normal fertile males revealed a pattern of over 200 proteins, ranging in mass from 10 000 to 100 000 daltons. Comparison of the mapped proteins from these males and those who had undergone vasectomy allowed us to identify one series of glycoproteins as missing from the semen from vasectomized individuals. Glycoproteins isolated by affinity chromatography with use of concanavalin A were also mapped. Some of the protein spots were identified either by co-electrophoresis with purified proteins or by the electrophoretic transfer of proteins to nitrocellulose sheets and subsequent detection by immunological procedures. The proteins identified include a number of serum proteins as well as prostatic acid phosphatase and creatine kinase. Proteolytic events shown to occur during the liquefaction of semen that occurs early after collection indicate the importance of carefully controlled collection and preparation methods for clinical evaluation of seminal plasma. Ethylenediaminetetraacetic acid and phenylmethylsulfonyl fluoride inhibit this proteolysis.

25. Two-dimensional analysis of human lymphocyte proteins: I. An assay for lymphocyte effectors. Willard, K. E. and Anderson, N. G. Clin Chem 27(8), 1327-34. (19 81).

We describe an assay for lymphocyte effectors that is capable of establishing the existence of regulators of lymphocyte gene expression (including post-transcriptional control and protein processing) and has the ability to characterize the response at the molecular level. The hypothesis that circulating effectors substances excreted through the kidney can be actively present in human urine was tested with this assay. Thus, biologically active protein molecules in urine were detected at concentrations of less than 1 mg/L and over a wide range of dilutions. Activities were detected and quantitated by culturing human lymphocytes with human urinary proteins in the presence of [35S]methionine and subsequently analyzing the labeled lymphocyte proteins by two-dimensional gel electrophoresis. Thus, protein analysis by two-dimensional gels was used to indirectly detect changes produced in cultured lymphocytes after exposure to regulatory molecules. Proteins or sets of lymphocyte proteins appeared or disappeared after exposure to normal or pathological human urinary proteins. Normal human urinary proteins triggered the appearance of sets of proteins referred to by number as the "Urocon" proteins and suppressed the synthesis of protein sets referred to as "Urocof" proteins. In addition to the normal alterations described, urinary proteins from individuals with influenza or acute leukemia and after renal transplantation were capable of inducing unique alterations in lymphocyte patterns.

26. Muscle protein analysis. II. Two-dimensional electrophoresis of normal and diseased human skeletal muscle. Giometti, C. S., Barany, M., Danon, M. J., and Anderson, N. G. Clin Chem 26(8), 1152-5. (1980).

We used high-resolution two-dimensional electrophoresis to analyze the major proteins of normal and pathological human-muscle samples. The normal human-muscle pattern contains four myosin light chains: three that co-migrate with the myosin light chains from rabbit fast muscle (extensor digitorum longus), and one that co-migrates with the light chain 2 from rabbit slow muscle (soleus). Of seven Duchenne muscular dystrophy samples, four yielded patterns with decreased amounts of actin and myosin relative to normal muscle, while three samples gave patterns comparable to that for normal muscle. Six samples from patients with myotonic dystrophy also gave normal patterns. In nemaline rod myopathy, in contrast, the pattern was deficient in two of the fast-type myosin light chains.

27. Analytical techniques for cell fractions. XXVII. Use of heart proteins as reference standards in two-dimensional electrophoresis. Giometti, C. S., Anderson, N. G., Tollaksen, S. L., Edwards, J. J., and Anderson, N. L. Anal Biochem 102(1), 47-58. (1980).

28. Alterations of gene expression in Novikoff hepatoma cells induced by a factor in human urine. Willard, K. E. and Anderson, N. G. Biochem Biophys Res Commun 91(3), 1089-94. (1979).

29. Analytical techniques for cell fractions. XXVI. A two-dimentional electrophoretic analysis of basic proteins using phosphatidyl choline/urea solubilization. Willard, K. E., Giometti, C. S., Anderson, N. L., O'Connor, T. E., and Anderson, N. G. Anal Biochem 100(2), 289-98. (1979).

30. Muscle protein analysis. I. High-resolution two-dimensional electrophoresis of skeletal muscle proteins for analysis of small biopsy samples. Giometti, C. S., Anderson, N. G., and Anderson, N. L. Clin Chem 25(11), 1877-84. (1979).

We have been developing a clinically useful method for high-resolution two-dimensional electrophoretic analysis of small (5--10 mg) human muscle biopsy samples with sufficient resolution to resolve the major contractile proteins and enzymes. Using rabbit psoas muscle as a model, we describe methods for sample preparation and two-dimensional electrophoresis. Basic proteins, which appear as streaks when conventional isoelectric focusing is used in the first dimension, are resolved through a modification of the nonequilibrium pH gradient electrophoresis method [Cell 12, 1133 (1977)]. In the two-dimensional patterns obtained from rabbit muscle, we identify the components of 10 enzymes and of myosin, actin, tropomyosin, and troponin. These patterns indicate charge heterogeneity in a large fraction of the proteins. Comparison of rabbit and normal human muscle patterns shows many similarities, but much additional work is required to confirm identifications. We conclude that analysis of small biopsy samples is feasible, but that all aspects of human sample acquisition, storage (when necessary), and preparation require thorough study before the method becomes routine in human muscle research and, ultimately, in the diagnosis of some muscle diseases.

31. Proteins of human urine. I. Concentration and analysis by two-dimensional electrophoresis. Anderson, N. G., Anderson, N. L., and Tollaksen, S. L. Clin Chem 25(7), 1199-210. (1979).

32. Red cell proteins. I. Two-dimensional mapping of human erythrocyte lysate proteins. Edwards, J. J., Anderson, N. G., Nance, S. L., and Anderson, N. L. Blood 53(6), 1121-32. (1979).

Human erythrocyte lysate proteins were resolved into over 250 discrete spots by two-dimensional electrophoresis using isoelectric focusing in the first dimension and electrophoresis in the presence of sodium dodecyl sulfate, (SDS) in the second. The overwhelming excess of hemoglobin has made such analyses difficult in the past. However, with the ISO-DALT two-dimensional electrophoresis system, large numbers of red cell proteins can be mapped in the presence of hemoglobin. When hemoglobin and several other major proteins are removed by adsorption to DEAE-cellulose, additional minor components are seen, giving a total of over 275. With the use of purified preparations, the map positions of five cell enzymes or their subunits were determined: pyruvate kinase, catalase, glucose-6-phosphate dehydrogenase, hypoxanthine phosphoribosyltransferase, and carbonic anhydrase. The mapping techniques described complement and extend those traditionally used to find human red cell protein variants.

33. Microheterogeneity of serum transferrin, haptoglobin and alpha 2 HS glycoprotein examined by high resolution two-dimensional electrophoresis. Anderson, N. L. and Anderson, N. G. Biochem Biophys Res Commun 88(1), 258-65. (1979).

34. Analytical techniques for cell fractions. XXV. Concentration and two-dimensional electrophoretic analysis of human urinary proteins. Anderson, N. G., Anderson, N. L., Tollaksen, S. L., Hahn, H., Giere, F., and Edwards, J. Anal Biochem 95(1), 48-61. (1979).

35. Analytical techniques for cell fractions. XXII. Two-dimensional analysis of serum and tissue proteins: multiple gradient-slab gel electrophoresis. Anderson, N. L. and Anderson, N. G. Anal Biochem 85(2), 341-54. (1978).

36. Analytical techniques for cell fractions. XXI. Two-dimensional analysis of serum and tissue proteins: multiple isoelectric focusing. Anderson, N. G. and Anderson, N. L. Anal Biochem 85(2), 331-40. (1978).

37. The future of clinical chemistry. Anderson, N. G. Am J Med Technol 44(3), 233-7. (1978).

38. High resolution two-dimensional electrophoresis of human plasma proteins. Anderson, L. and Anderson, N. G. Proc Natl Acad Sci U S A 74(12), 5421-5. (1977).

The two-dimensional electrophoretic technique of O'Farrell has been adapted to the analysis of human plasma proteins, and 30 polypeptides have been identified in the pattern produced. Genetic variants involving charge (isoelectric point) or size (molecular weight in the presence of sodium dodecyl sulfate) changes should be routinely detectable in at least 20 proteins at once, facilitating studies of human mutation rates.

OTHER SUBJECTS

1. Anderson, Norman G. Embryonalentwicklung, Retrogenese und Krebs. (1975). Mainz
Wiesbaden, Akademie der Wissenschaften und der Literatur ;
Steiner in Kommission.

2. Anderson, Norman G., Townsend, J. Ives, Society of the Sigma Xi. University of Tennessee Chapter., and Oak Ridge National Laboratory. Biology Division. Lectures in biological sciences. (1963). Knoxville, University of Tennessee Press.

3. Anderson, Norman G., Coggin, Joseph H., and Oak Ridge National Laboratory. Proceedings of the first [-second] Conference and Workshop on Embryonic and Fetal Antigens in Cancer. (1971). [Washington] , U.S. Atomic Energy Commission; [available from the National Technical Information Service, Springfield, Va.

4. Large-scale oligonucleotide synthesizers. I. Basic principles and system design. Anderson, N. G., Anderson, N. L., Taylor, J., and Goodman, J. Appl Biochem Biotechnol 54(1-3), 19-42. (1995).

The central problem in scaling up oligonucleotide synthesis is to expose each element of a large bed to the same conditions obtaining in very small ones, for the same intervals of time. Our analysis suggests that scale-up is chiefly limited by fluid path length through the bed. By using annular beds in zonal centrifuge rotors of unique design, with fluid flow controlled by combining centrifugal force with differences in physical density between reagents, reagent fronts may be kept exactly perpendicular to the direction of flow in each bed element. Under these conditions, bed volume may be increased by increasing rotor length and diameter. The rotor is lined with polypropylene or Teflon, and has a thick tempered glass end window. Transparent rotary valves of a unique design allow any of 47 reagents to be selected and the direction of flow through the rotor to be controlled. A photodiode spectrophotometer provides complete absorption spectra on fluid in the rotor inlet and outlet lines every 4 s, and a large balance weighs effluent from the synthesizer continuously. The entire operation is controlled by a work station, and steps may be programmed by time, absorbance, or reagent mass. Reagents are identified by spectra, and trityls are integrated on line. A detailed time-stamped log file provides a complete record of each synthesis.

5. Fetal antigens shared as transplantation rejection antigens on chemically induced mouse and hamster sarcomas. Coggin, J. H. Jr., Adkinson, L., and Anderson, N. G. Cancer Res 40(5), 1568-73. (1980).

6. Construction of tygon manifolds and connectors. Anderson, N. G. Anal Biochem 86(1), 337-8. (1978).

7. Molecular basis for programming in development. Introduction. Anderson, N. G. and Coggin, J. H. Jr. Cancer Res 36(9 PT 2), 3384-6. (1976).

8. The effect of exposure to dibutyryl cyclic adenylic acid on the membrane antigenicity of SV40 tumor cells. Hannon, W. H., Anderson, N. G., and Coggin, J. H. Jr. Cancer Res 36(9 PT 2), 3429-34. (1976).

When SV40 tumor cells (F5-1) in culture were treated with dibutyryl cyclic adenylic acid (DbcAMP),marked alterations occurred in their growth and morphology. Additionally, the incorporation or uptake of labeled thymidine, uridine, phenylalanine, and choline were reduced by this treatment. These modifications with DbcAMP exposure produced conditions simulating those of a contact-inhibited state. The immunization of hamsters with X-irradiated tumor cells previously cultured in the presence of DbcAMP indicated that the tumor-specific transplantation antigens and/or fetal antigens were altered during short-term expsoure to DbcAMP. Further examination of membrane antigens of SV40 tumor cells using the isotopic antiglobulin technique demonstrated a significant reduction in the tumor membrane antigen on cells cultured with DbcAMP.

9. Interactive macromolecular sites. II. Role in prebiotic macromolecular selection and early cellular evolution. Anderson, N. G. J Theor Biol 60(2), 413-9. (1976).

10. Interactive macromolecular sites. I. Basic theory. Anderson, N. G. J Theor Biol 60(2), 401-12. (1976).

11. Early detection of pregnancy-associated serum protein using antiserum to placental antigens. Holladay, D. W., Caton, J. E., Ball, F. L., Holleman, J. W., and Anderson, N. G. Immunol Commun 5(1-2), 1-11. (1976).

Antisera against human placental proteins were developed in goats and rabbits, using immunoadjuvants and a prolonged injection schedule. The antisera were absorbed with normal serum proteins and then tested in immunodiffusion against normal and pregnancy sera. Two bands of precipitation due to pregnancy antigens were observed in pregnancy sera as early as 18 days after conception. Detection of these antigens has possibilities for application as an early pregnancy test.

12. Analytical techniques for cell fractions. XX. Cyclic affinity chromatography: principles and applications. Anderson, N. G., Willis, D. D., Holladay, D. W., Caton, J. E., Holleman, J. W., Eveleigh, J. W., Attrill, J. E., Ball, F. L., and Anderson, N. L. Anal Biochem 68(2), 371-93. (1975).

13. Analytical techniques for cell fractions. XIX. The cyclum: an automatic system for cyclic chromatography. Anderson, N. G., Willis, D. D., Holladay, D. W., Caton, J. E., Holleman, J. W., Eveleigh, J. W., Attrill, J. E., Ball, F. L., and Anderson, N. L. Anal Biochem 66(1), 159-74. (1975).

14. Proceedings: Third Conference of Embryonic and Fetal Antigens in Cancer. Introduction. Anderson, N. G. and Coggin, J. H. Jr. Cancer Res 34(8), 2032-3. (1974).

15. Proceedings: Searching for human tumor antigens. Anderson, N. G., Holladay, D. W., Caton, J. E., Candler, E. L., Dierlam, P. J., Eveleigh, J. W., Bail, F. L., Holleman, J. W., Breillatt, J. P., and Coggin, J. H. Jr. Cancer Res 34(8), 2066-76. (1974).

16. Proceedings: Proposed mechanisms by which autochthonous neoplasms escape immune rejection. Coggin, J. H. Jr., Ambrose, K. R., Dierlam, P. J., and Anderson, N. G. Cancer Res 34(8), 2092-101. (1974).

17. Engineering versus disease. Anderson, N. G. Ann Biomed Eng 2(1), 1-18. (1974).

18. Cancer, differentiation and embryonic antigens: some central problems. Coggin, J. H. Jr. and Anderson, N. G. Adv Cancer Res 19(0), 105-65. (1974).

19. Molecular mechanisms in blocked ontogeny and retrogenesis. Anderson, N. G. and Coggin, J. H. Jr. Ann N Y Acad Sci 230, 508-15. (1974).

20. Phase specific surface autoantigens on membranes of fetus and tumors. Coggin, J. H. Jr., Ambrose, K. R., and Anderson, N. G. Adv Exp Med Biol 29(0), 483-90. (1973).

21. Development of a miniature fast analyzer. Burtis, C. A., Mailen, J. C., Johnson, W. F., Scott, C. D., Tiffany, T. O., and Anderson, N. G. Clin Chem 18(8), 753-61. (1972).

22. Variable-speed drive for the mechanical stage of a microscope used as the optical component in a microdensitometer. Caton, J. E., Willis, D. D., and Anderson, N. G. Anal Biochem 46( 1), 232-8. (1972).

23. Cytostatic antibody and SV40 tumour immunity in hamsters. Ambrose, K. R., Anderson, N. G., and Coggin, J. H. Jr. Nature 233(5318), 321-4. (1971).

24. Interruption of SV40 oncogenesis with human foetal antigen. Ambrose, K. R. , Anderson, N. G., and Coggin, J. H. Nature 233(5316), 194-5. (1971).

25. Tumor immunity in hamsters immunized with fetal tissues. Coggin, J. H. Jr., Ambrose, K. R., Bellomy, B. B., and Anderson, N. G. J Immunol 107(2), 526-33. (1971).

26. Analytical techniques for cell fractions. 18. Use of cellulose wicks to monitor agglutination reactions. Anderson, N. G. Anal Biochem 38(1), 175-89. (1970).

27. Fetal antigen capable of inducing transplantation immunity against SV40 hamster tumor cells. Coggin, J. H., Ambrose, K. R., and Anderson, N. G. J Immunol 105(2), 524-6. (1970).

28. Analytical differential centrifugation: an analysis of the sedimentation properties of synaptosomes, mitochondria and lysosomes from rat brain homogenates . Cotman, C., Brown, D. H., Harrell, B. W., and Anderson, N. G. Arch Biochem Biophys 136(2), 436-47. (1970).

29. Induction of tumor-specific transplantation immunity in hamsters with cell fractions from adenovirus and SV40 tumor cells. Coggin, J. H., Elrod, L. H., Ambrose, K. R., and Anderson, N. G. Proc Soc Exp Biol Med 132(1), 328-36. (1969).

30. Analytical techniques for cell fractions. X. High-pressure ninhydrin reaction system. Anderson, N. G., Stevens, R. H., and Holleman, J. W. Anal Biochem 26(1), 104-17. (1968).

31. Lipid class and fatty acid composition of rat liver plasma membranes isolated by zonal centrifugation. Pfleger, R. C., Anderson, N. G., and Snyder, F. Biochemistry 7(8), 2826-33. (1968).

32. Analytical techniques for cell fractions. IX. Measurement and transfer of small fluid volumes. Anderson, N. G. Anal Biochem 23(2), 207-18. (1968).

33. Automatic, high-resolution analysis of urine for its ultraviolet-absorbing constituents. Scott, C. D., Attrill, J. E., and Anderson, N. G. Proc Soc Exp Biol Med 125(1), 181-4. (1967).

34. High-pressure column chromatography. I. Design of apparatus and separation of bases, nucleosides, and nucleotides. Green, J. G., Nunley, C. E., and Anderson, N. G. Natl Cancer Inst Monogr 21, 431-40. (1966).

35. Centrifugal freezing. I. A system for rapid freezing of aqueous cell suspensions. Anderson, N. G., Green, J. G., and Mazur, P. Natl Cancer Inst Monogr 21, 415-30. (1966).

36. The isolation of oral structures from Tetrahymena pyriformis by low-speed zonal centrifugation. Whitson, G. L., Padilla, G. M., Canning, R. E., Cameron, I. L., Anderson, N. G., and Elrod, L. H. Natl Cancer Inst Monogr 21, 317-21. (1966).

37. Studies on synchronized cells: radiation-induced division delay in the flagellate Astasia longa. Padilla, G. M., van Dreal, P. A., and Anderson, N. G. Radiat Res 28(1), 157-65. (1966).