Dr. S. DeCarvalho, Laboratory of Haematology, Doctors Hospital and Laboratory of Cancer Virology and Immunology, Rand Development Corporation, Cleveland, Ohio. 

It is now generally accepted that RNA, conveying DNA's information, directs specific proteinopoiesis. This notion, derived mainly from cytochemical and biochemical evidence, is being utilized to demonstrate specific proteinopoietic activities of native ribonucleic acids extracted from different cellular systems. It has been shown that RNA from yeast cells adapted to copper specifically protects unadapted strains against lethal doses of copper (1), and that a new type of RNA appears in yeast cells when catalase synthesis is induced by exposure to oxygen (2).

True redifferentiation of mouse ascites tumour cells by RNA from normal calf liver may have been obtained by Niu (3-5). On the other hand, normal mouse kidney cells treated by RNA from ascites tumour cells were able to induce local tumours in mice (3). We have shown that RNA from human leukaemic and tumour cells can be cytopathogenic for cultures of primary lines of human cells (6, 7), and oncogenic and leukaemogenic for animals (8). The interplay of normal and malignant RNA with malignant and normal cells obtained with the Novikoff tumour model points towards the possibility of redifferentiating human malignant cells with human normal RNA (9-12). RNA information transfer between sickle haemoglobin -producing and normal human erythroblasts was apparently obtained by Weisberger (13). Significant reduction in tumour takes of normal mouse liver were obtained by Harel et al (14) by the effect of normal mouse liver RNA on FLS ascites carcinoma cells of the Swiss mouse. Proof of penetration of labelled RNA into normal and tumour cells was obtained by Schwarz et al (31).

The foregoing evidence led to the hypothesis that RNA from malignant cells is biologically different from RNA from non-malignant cells and that the degree of differentiation or undifferentiation of the malignant cells might reflect the relative amounts of normal and abnormal RNA. Abnormal RNA would be unable to direct specific protein synthesis; enzyme deletions and antigenic simplification, both found in malignant cells, could be the indirect result of the unfitness of the abnormal RNA (15). Of the two classes of RNA now described by Sibatani (16) the high metabolically active, low in guanidine content and the less metabolically active, high in guanidine content, the last was reported a few years ago to be predominant in malignant cells (17).

A pattern for alterations of normal RNA under carcinogenic influences (viral, physical, chemical) may have been set by the experiments of Tsugita and Fraenkel-Conrat (18) and Siegel (19), and the findings of Alderson (20). The transformation of cytosine into uracil in the RNA of the tobacco mosaic virus by nitrous acid resulted in a mutant producing a protein differing from the natural TMV protein in the replacement of proline by leucine in a certain location. It is recalled that a similar difference in one single amino-acid is all that distinguishes haemoglobin A from sickle haemoglobin and other haemoglobins. Similar mutagenic alterations on the TMV RNA were brought about by 4-nitroquinoline-N-oxide (32), a substance which is leukaemogenic for the mouse by mechanisms suggesting endovirogenesis (33).

Departing from the hypothesis that malignant cells remain undifferentiated because of a deficiency of normal proteinopoietic RNA, an attempt was made to supply these cells in situ,  with RNA extracted from normal cells. Leukaemic marrow and RNA from normal bone marrow seemed an ideal system for such experiment.

It must be emphasized that the aim of the work recorded here is to examine some biological properties of the RNA from normal marrow, specifically to determine if the haemopoietic information it contains can be transferred by a sort of sub-cellular homografting of a template to other cells. This work by no means contains a therapeutic or pharmacological implication, and it is presented for the possible biological consequences rather than for its content of clinical repercussions.

RNA from Human Normal Bone Marrow:

Non-leukaemic bone marrow was obtained from a 48-year-old white male presenting a history of epigastric distress which on X-ray was shown to be due to herniation of gastric mucosa through the pylorus. The following were his haematological parameters: Haematocrit value 52; haemoglobin 15.30 g per cent; red blood cells 5,200,000 per mm³; white blood cells 8,000 per mm³;differential count in the peripheral blood: neutrophiles 57 per cent, lymphocytes 42 per cent, and monocytes 1 per cent.

25 ml. of marrow were drawn from a sternal puncture in 2.5 ml. of 3.8 per cent sodium citrate. This marrow contained a total of 1.75 x 10⁹ cells per ml. and the myelogram was within normal limits.

RNA from this marrow was extracted by phenol. The alcohol-precipitate was redisolved in isotonic phosphate-buffered saline at pH 7.1. The final concentration of RNA was adjusted to 90 mg/ml. This preparation was filtered through 03 Selas porcelain filters and kept at -20^o C for 24 h when it was thawed to 2^o C for use.

Leukaemic Patients' Histories:

H.M., 57-year-old white male. Admitted for the first time on October 28, 1959, because of petechiae and small ecchymoses over chest and limbs. Three weeks before admission started noting easy fatigue and tiredness, pallor of skin and mucosae, petechiae and small ecchymoses and papular infiltrations over the skin of chest and legs. His blood data were: haemoglobin 8.65 g per cent; red blood cells 2,560,000/mm³; white blood cells 11,400/mm³; 52 per cent stem cells, 30 per cent lymphocytes, 11 per cent neutrophiles, 1 per cent plasmocytes; 40,000/mm³ platelets, 0 reticulocytes. The sternal bone marrow was hypoplastic and paucicellular with partial megaloblastic transformation of the erythropoiesis. The leukemic cells, representing about 15 per cent of all elements, were of myeloblastoid resemblance.

The treatment from October 30, 1959, to March 14, 1960 had consisted of transfusion, methyl- prednisolone and 6-mercaptopurine.

During the period of treatment the haemoglobin oscillated between 8 and 12 g per cent, the leukocyte count from 14 to 40,000/mm³ and the platelets from 40 to 80,000/mm³ with 25-90 per cent of leukaemic cells. No remissions were ever achieved, although the general condition was satisfactory,the patient was asymptomatic, ambulatory and even resumed part-time work. Two weeks before this experiment, all treatment was discontinued. Tables 1 and 2 contain marrow data a few minutes before the experiment started.
 
 
 

Table 1. Myelogram of Patient H.M. (% of 500 cells counted)

Place of  instillation	Before normal RNA	2 days after 1st  instillation of normal RNA	10 days after normal RNA
Sternum	Leukaemic cells-- 84	63	91.5
.	Erythroblasts------ 9	21	6
.	Myelocytes---------7	14.5	2.0
.	Megakaryocytes--0	1.5	0.5
Place of instillation	Before buffered saline	2 days after buffered saline	10 days after buffered saline
Iliac bone	Leukaemic cells--92.5	92	92
.	Erythroblasts-------5	6.5	5.5
.	Myelocytes---------2.5	1.5	2.5
.	Megakaryocytes---0	0	0

M.H., a 71-year-old white male. Admitted on April 1, 1960, because of weakness, dizziness and faintness. The laboratory findings on admission were : haemoglobin 7.5 g per cent; red blood cells 2,160,000/mm³; white blood cells 4,100/mm³; of the last, 50 per cent were myeloblastoid anaplastic stem cells and 5 per cent basophils. The bone marrow was cellular and monomorphic, 95 per cent of its cells being leukaemic cells. Treatment consisted, until 2 weeks before this experiment, of transfusions and antibiotics.
 
 
 

.Table 2. Myelograms of Patient M.H. (% of 500 cells counted).

Place of  instillation	Before normal RNA	2 days after normal RNA	10 days after normal RNA
Sternum	Leukaemic cells----99	55	99
.	Erythroblasts---------1	30	0.5
.	Myelocytes-----------0	13.5	0.5
.	Megakaryocytes----0	1.5	0
Place of instillation	Before buffered saline	After buffered saline	10 days after buffered saline
Iliac bone	Leukaemic cells---99.5	99	100
.	Erythroblasts--------0.5	0.5	0
.	Myelocytes-----------0	0.5	0
.	Megakaryocytes----0	0	0

Intra-Osseous Instillation of Normal Marrow RNA:

A puncture was made in the middle of the second sternal piece. Marrow was aspirated for morphological study. Through the same needle 1 ml. of the normal RNA preparation was injected. At the same time a puncture was made in the right iliac crest at a point 1 cm behind the anterior superior spine at an angle pointing to this spine. Marrow was obtained for morphological study followed by injection of 1 ml. of phosphate-buffered saline as control. Later aspirations of marrow were made in the same marked places.

Differential counts represent percentages of cells in Wright stained smears. Each value is the average of three different counts. Absolute total counts were made from a suspension of 2 ml. of marrow diluted to 10 ml. with citrate-buffered saline. Counts were made in Levy-Hausser counting chambers. Each value represents an average of three counts in 64 large squares. Absolute figures for leukaemic cells, myelocytes and megakaryocytes were calculated from the percentages in smears and total counts. Absolute counts of erythroblasts were obtained directly  in cell suspensions with the violet light microscope method described by Wilkins and us (21, 22) making use of the specific haem absorption in the Soret band. This method permitted to check the accuracy of the indirect absolute estimations which we found to represent an approximation of +/- 7 per cent.

Tables 1 and 2 summarize the cellular composition of the marrows obtained before and after instillation of normal marrow RNA and control-buffered saline.

Because of the slight, though clear, change in the haemopoietic direction and the quick reverse to the previous full leukaemic stage after this first RNA instillation, an amount of normal marrow RNA 10 times larger was reinstilled in patient H.M. 900 mg of the same RNA in 10 ml. of buffered saline was instilled in the sternal manubrium 30 days after the first RNA instillation in the second sternal piece. Samples of marrow were taken before and 2, 6, 10, and 14 days after this reinstillation. Table 3 gives the results.
 
 

Table 3. Relative and Absolute Differential Cell Counts of Marrow Cells of Patient H.M. after Second Reinstillation of 900 mg of Normal Human Marrow in the Sternal Manubrium.

Cells	Time after  RNA instillation	.	.	.	.	.	.	.	.	.
.	0  days	.	2  days	.	6  days	.	10  days	.	14 days	.
.	Rel.	Abs.	Rel.	Abs.	Rel.	Abs.	Rel.	Abs.	Rel.	Abs.
Leukaemic Cells	91	3.7	2	0.1	20	0.6	40	1.2	90	3.6
Erythro- blasts	6.5	0.25	65	2	56	1.6	36	1	6	0.2
Myelo- cytes	1.5	0.1	31	0.9	22	0.6	13	0.4	4	0.2
Megakaryo- cytes	0	0	2	0.6	2	0.1	0.3	0.001	0	0
Total absolute cell numbers	4.05	.	3.6	.	2.9	.	2.6	.	4.0	.

Relative counts: Percentages in 1,000 cells in smears.
Absolute counts: Millions in 2 ml. of marrow. 

Morphological studies and differential cell counts of the marrows of the two leukaemic patients clearly indicated a local haemopoietic response following instillation of the RNA from normal marrow. For reasons not well understood, this haematopoietic response either fades away or it is completely obscured by the leukaemic proliferation. There seems to be a relationship between the amount of RNA instilled and the degree of haemopoietic response as well as the period of time for reversion to the previous leukaemic stage. When 90 ug of RNA were instilled, only a slight haemopoietic response was noted with reversion to the statu quo ante within 10 days. When 10 times that amount was re-instilled, a full haemopoietic response was obtained in 2 days with progressive decline within 14 days.

Failure of intravenous administration of RNA to induce marrow changes may probably be accounted for by quick inactivation in the circulation by ribonuclease or inhibition by strong combination of the RNA with plasma proteins. Data on this problem are apparently conflicting. Gierer and Schramm (23) have indicated that highly purified RNA from tobacco mosaic virus is largely inactivated by proteins in serum. However, injections of tumoral and leukaemic RNA have been successful in inducing disease in mice (8, 24-26), thus indicating persistence of activity after injection. It may be that these preparations are not so highly purified as those of Schramm and that residual protein affords some degree of protection. Ellem (27) and we found (7) that in testing the cytopathogenicity of RNA preparations, it is very important to wash off the medium of the tissue culture and to operate in conditions of osmolality, pH, temperature and time for absorption which proved the most favourable to the nucleic acid, while extracellular. The foregoing conditions were to a certain extent reproduced in the method of intra-osseous instillation and this fact probably accounts for the success of local application and the failure of the intravenous administration of the RNA.

While it is possible to obtain these conditions in vitro, it will probably be necessary to learn how to reproduce them in vivo before we will be able to bioassay specific proteinopoietic activities of the ribonucleic acids from different sources.

A third patient provided the opportunity for further probing into the possibilities of protecting RNA intravenously injected. W.B., a 40-year-old male, was diagnosed of acute stem-cell leukaemia about three months after the first symptoms of weakness and fatigue. The patient had not received any form of treatment, not even transfusions. By the time of the experiment the haematological findings were: sternal bone marrow: 95 per cent leukaemic cells; haemoglobin 7.00 g per cent; red blood cell 2.8 x 10⁶/mm³; white blood cell 4 x 10⁴/mm³ of which 80 per cent were leukaemic cells, reticulocytes 0 per cent, platelets 2 x 10⁴/mm³.

An intravenous infusion of 1 g of calcium disodium versenate in 150 ml. of 5 per cent glucose in water was dripped for 1 h. Urine collected before, during this time, and in the following 2 h was assayed for calcium content, and showed an elimination of 1/5 of the total previous blood calcium. Blood calcium-levels, however, did not show appreciable differences due to bone calcium mobilization in accordance with reported findings (28). Magnesium in the blood dropped from 3 to 0.7 mg per cent.

Immediately following the last drops of the chelating agent 1,480 mg of RNA in buffered versanate saline obtained from pooled normal human bone marrow were injected intravenously.

The same procedure was repeated for 3 days. 1-2 h after the EDTA-RNA administration, chills and temporary elevation of temperature were noted. Table 4 summarizes the haematological findings in the first 2 weeks of observation.
 
 

Table 4. Effect of Intravenous EDTA Followed by Normal Marrow RNA on Leukaemic Blood.

Day	Hb	RBC	WBC	Leukaemic cells	Platelets	Reticulo- cytes
.	(g%)	106 /mm3	103 /mm3	% of WBC	104 /mm3	(%)
0	7.0	2.8	50	80	2	0
0.5	7.0	2.8	57	85	2	0
1	7.0	3.0	28	40	4	0.1
2	7.8	3.2	18	30	5	1.0
2.5	.	.	.	.	.	.
3	8.10	3.4	12	10	6	1.4
3.5	.	.	.	.	.	.
4	8.80	3.5	12	10	6	2.7
5	9.1	3.7	12	10	6	3.1
6	10.1	3.8	10	6	6	4
7	10	3.9	10	6	6	4.2
15	8	3.2	26	15	4	3.1

The haematopoietic response by the systemic RNA, as shown by the moderate reticulocyte elevation and haemoglobin build up, was maintained for about the same time as that observed in the local stimulation by regional instillation of RNA.

The question of whether the phenomena described represent specific haemopoietic redifferentiation of the leukaemic cells by a haemopoietic RNA template or if they represent non-specific haemopoietic stimulation of reticulum cells by the preparation cannot be solved until RNA from organs other than bone marrow is used in the same way. Local changes of megaloblastic into normoblastic marrow were induced by Horrigan (29) following local instillation of vitamin B₁₂. In 1927 Larsell et al (30) injected nucleic acid preparations from erythrocytes of the fowl intravenously into rabbits and into 11 anaemic patients. Marked haemopoietic stimulus was noted as judged by peripheral blood cell cuts. With the method of preparing the nucleic acids used by these authors, degradation of the nucleic acids and heavy protein contamination was to be expected and thus transfer of intact templates may probably be excluded. Non-specific stimulation of the haemopoiesis by thymidine residues and analogues was probably the mechanism of action in this case. This and similar works established the basis for the therapeutic use, now discontinued, of nucleotides in aplastic anaemia to stimulate haemopoiesis. However, one problem is to stimulate haemopoiesis and another is to redifferentiate anaplastic leukaemic stem cells. Further evidence for true redifferentiation of the leukaemic cells may have to be gained by the use of normal RNA on cultures of leukaemic cells in the absence of potentially haemopoietic normal reticulum cells.

References:

1. Minigawa T, Biochim. Biophys. Acta 16: 539 (1955).

2. Chantrenne H, Arch. Biochim. Biophys. 65: 414 (1956).

3. Niu MC, Cordova CC, and Niu LC, Proc. Natl. Acad. Sci. U.S.A. 47: 1689 (1961).

4. Niu MC, Cordova CC, Niu LC, and Radbill CL, Proc. Natl. Acad. Sci. U.S.A. 48: 1964 (1962).

5. Niu MC, "Eighth Intern. Cancer Cong.", 110, (Medgiz Publishing House, Moscow, 1962).

6. DeCarvalho S, "Proc. Third Canad. Cancer Conf.", 329 (Academic Press, New York, 1959).

7. DeCarvalho S, J. Lab. Clin. Med., 55: 694 (1960).

8. DeCarvalho S, J. Lab. Clin. Med., 55: 706 (1960).

9. DeCarvalho S, and Rand HJ, Nature 189: 815 (1961).

10. Griffin AC, and O'Neal MA, Eighth Intern. Cancer Cong. 111, (Medgiz Publishing House, Moscow, 1962).

11. O'Neal MA, Ward V, and Griffin AC, Proc. Amer. Assoc. Cancer Res. 3: (March, 1962).

12. Askenova NN, Bresler VM, Vorobyev VI, and Olenov JM, Nature 196: 443 (1962).

13. Weisburger AS, Proc. Natl. Acad. Sci. U.S.A. 48: 68 (1962).

14. Harel J, Lacour F, and Verger C, Cancer Chemother. Abst. 3: 592 (October, 1962).

15. Busch H, "An Introduction to the Cytochemistry of the Cancer Cell", (Academic Press, New York, 1962).

16. Sibatani A, Yamana K, Kimura K, and Takashashi T, Nature 186: 215 (1960).

17. DeLamirande G, Allard C, and Cantero A, Cancer Res. 15: 329 (1955).

18. Tsugita AK, and Fraenkel-Conrat H, Proc. Natl. Acad. Sci. U.S.A., 47: 636 (1960).

19. Siegel A, Virology 11: 156 (1960).

20. Alderson T, Nature 185: 905 (1960).

21. Wilkins MHF, and DeCarvalho S, Blood 8: 944 (1953).

22. DeCarvalho S, Blood 10: 453 (1955).

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25. Hays EF, Simmons NS, and Belk WS, Nature 180: 1419 (1957).

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33. Kinosita R, in "Viruses, Nucleic Acids and Cancer, Seventeenth Symp. Fundamental Cancer Res. (Univ. Texas, in the press); and personal communications (1962,1963).

Additional References for RNA as a Therapeutic Agent:

1. DeCarvalho S, and Rand HJ, "Comparative Effects of Liver and Tumour Ribonucleic Acids on the Normal Liver and the Novikoff Hepatoma Cells of the Rat", Nature, vol. 189, no. 4767, pp. 815-817 (March 11, 1961).

2. Niu MC, Cordova CC, and Niu LC, "Ribonucleic Acid-Induced Changes in Mammalian Cells", Proc. Natl. Acad. Sci. USA, vol. 47, pp. 1689-1700 (October, 1961).

3. DeCarvalho S, "Effect of RNA from Normal Human Marrow on Leukaemic Marrow In-Vivo", Nature, vol. 197, no. 4872, pp. 1077-1080 (March 16, 1963).

4. Frenster JH, "Nuclear Polyanions as De-Repressors of Synthesis of Ribonucleic Acid", Nature, vol. 206, no. 4985, pp. 680-683 (May 15, 1965).

5. Czihak G, "Evidence for inductive properties of the micromere RNA in sea urchin embryos",
Naturwissenschaften vol. 52, no. 6, pp.141-142 (1965).

6. Kronenberg LH, and Humphreys T, "Double-Stranded Ribonucleic Acid in Sea Urchin Embryos", Biochemistry vol. 11, no. 11, pp. 2020-2026 (1972).

7a. Kolodny GM, "Evidence for transfer of macromolecular RNA between mammalian cells in culture", Exp. Cell Res. vol. 65, no. 2, pp. 313-324 (April, 1971).

7b. Kolodny GM, "Cell to cell transfer of RNA into transformed cells", J. Cell Physiol. vol. 79, no. 1, pp. 147-150 (February, 1972).

7c. Kolodny GM, Culp LA, and Rosenthal LJ, "Secretion of RNA by normal and transformed cells", Exp. Cell Res. vol. 73, no. 1, pp. 65-72 (July, 1972).

8. Torelli U, Torelli G, and Cadossi R, "Double-Stranded Ribonucleic Acid in Human Leukemic Blast Cells", Europ. J. Cancer vol. 11, no. 2, pp. 117-121 (February, 1975).

9. Frenster JH, and Herstein PR, "RNA in Gene De-Repression", in: Proc. AAAS Symposium, December 28-30, 1972, "The Role of RNA in Reproduction and Development", edit. by Niu MC, and Segal SJ, pp. 330-338, North-Holland Publishing Co. Amsterdam, 1973.

10. Kern DH, Chow N, and Pilch YH, "Lymphocyte Populations Participating in Cellular Antitumor Immune Responses Mediated by Immune RNA", J. Natl. Cancer Inst. 60, 335-344 (1978).

11. Frenster JH, "Oncogenes as Molecular Targets within Active Chromatin", in: AACR-NCI-EORTC International Conference: "Molecular Targets and Cancer Therapeutics: Discovery, Development, and Clinical Validation", Washington, DC, November 16-19, 1999, and Published in: Clinical Cancer Research, vol. 5, suppl. l, p. 3855s, (624), (November, 1999).

12. Frenster JH, "Activation of DNA Transcription within Repressed Chromatin", 14th John Innes Symposium, September 5-8, 2001.

13. Maruo S, Nanbo A, and Takada K, "Replacement of the Epstein-Barr Virus Plasmid with the EBER Plasmid in Burkitt's Lymphoma cells", J. Virology, vol. 75, no. 20, pp. 9977-9982 (October, 2001).

14. Frenster JH, "Yeast  RNA  Re-Programming  of  Already-Active  Mammalian Chromatin", RNA2002, p. 592, (June, 2002).

15. Lanz RB, Chua SS, Barron N, Söder BM, DeMayo F, and O'Malley BW, "Steroid Receptor RNA Activator Stimulates Proliferation as Well as Apoptosis In Vivo", Mol. Cell. Biol., vol. 23, no. 20, pp. 7163-7176 (October, 2003).

16. Park W-S, Hayafune M, Miyano-Kurosaki N, and Takaku H, "Specific HIV-1 env gene silencing by small interfering RNAs in human peripheral blood mononuclear cells", Gene Therapy,  vol. 10, no. 24, pp. 2046-2050 (November, 2003).

17. Santulli-Marotto S, Nair SK, Rusconi C, Sullenger B,  and Gilboa E, "Multivalent RNA Aptamers That Inhibit CTLA-4 and Enhance Tumor Immunity", Cancer Research 63, 7483-7489, November 1, 2003.

18. Coughlin CM, Vance BA, Grupp SA, and Vonderheide RH, "RNA-transfected CD40-activated B cells induce functional T cell responses against viral and tumor antigen targets: implications for pediatric immunotherapy".

19. Davidson BL, "Hepatic Diseases --- Hitting the Target with Inhibitory RNAs", New Eng. J. Med., vol. 349, no. 24, pp. 2357-2359 (December 11, 2003).

20. Geiss G, Jin G, Guo J, Bumgarner R, Katze MG, and Sen GC, "A Comprehensive View of Regulation of Gene Expression by Double-Stranded RNA-Mediated Cell Signaling", J. Biol. Chem. vol. 276, pp. 30178-30182 (2001).

21. Persengiev SP, Zhu X and Green MR, "Nonspecific, concentration-dependent stimulation and repression of mammalian gene expression by small interfering RNAs (siRNAs)",  RNA, vol. 10, no. 1, pp. 12-18 (January, 2004).

22. Sayer NM, Cubin M, Rhie A, Bullock M, Tahiri-Aloui A , and James W , "Structural Determinants of Conformationally Selective, Prion-Binding Aptamers", J. Biol. Chem. vol. 279, no. 13, pp. 13102-13109 (March 26, 2004).

23. Mathas S, Lietz A, Anagnostopoulos I, Hummel F, Wiesner B, Janz M, Jundt F, Hirsch B, Jöhrens-Leder K, Vornlocher H-P, Bommert K, Stein H, and Dörken B, "c-FLIP Mediates Resistance of Hodgkin/Reed-Sternberg Cells to Death Receptor–induced Apoptosis",  J. Exp. Med., vol. 199, no. 8, pp. 1041-1052 (April 19, 2004).

24. Kuwabara T, Hsieh J, Nakashima K, Taira K, and Gage FH, "A Small Modulatory dsRNA Specifies the Fate of Adult Neural Stem Cells", Cell, vol. 116, no. 6, pp.779-793  (19 March 2004).

25. Takamizawa J, Konishi H, Yanagisawa K, Tomida S, Osada H, Endoh H, Harano T, Yatabe Y, Nagino M, Nimura Y, Mitsudomi T, and Takahashi T, "Reduced Expression of the let-7 MicroRNAs in Human Lung Cancers in Association with Shortened Postoperative Survival".

26. Frenster JH, and Hovsepian JA, "Activator RNA Exchange during Interphase Chromatin Reprogramming".

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Perspectives in atheroprophylaxis.
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Oncology. 1973;27(1):3-29. Review. No abstract available.
PMID: 4566134 [PubMed - indexed for MEDLINE]

9:  DeCarvalho S.
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unexplored features of fetal-maternal isoimmunity, longest recorded survival
following use of leukemostatic maternal isoantibody.
Oncology. 1973;27(1):52-63. No abstract available.
PMID: 4264687 [PubMed - indexed for MEDLINE]

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Nature. 1964 Sep 12;203:1187-8. No abstract available.
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Nature. 1964 Apr 18;202:255-8. No abstract available.
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PMID: 14026289 [PubMed - OLDMEDLINE for Pre1966]

15:  DECARVALHO S, RAND HJ.
 Antigens in human tumors revealed in suppressed rabbits rendered tolerant to
normal human tissues.
Exp Mol Pathol. 1963 Feb;2:32-9. No abstract available.
PMID: 14026288 [PubMed - OLDMEDLINE for Pre1966]

16:  DECARVALHO S, RAND HJ, UHRICK JR.
 Differences in information content of ribonucleic acids from malignant tissues
and homologous organs as expressed by their biological activities.
Exp Mol Pathol. 1962 Apr;1:96-103. No abstract available.
PMID: 13884719 [PubMed - OLDMEDLINE for Pre1966]

17:  DECARVALHO S, RAND HJ.
 Effects of iso- and hetero-antibodies against specific antigens of the
transplantable Novikoff hepatoma of the rat (solid form).
Nauchni Tr Vissh Med Inst Sofiia. 1962 Mar 10;193:950-2. No abstract available.
PMID: 13884720 [PubMed - OLDMEDLINE for Pre1966]

18:  DECARVALHO S, RAND HJ.
 Comparative effects of liver and tumour ribonucleic acids on the normal liver
and the Novikoff hepatoma cells of the rat.
Nature. 1961 Mar 11;189:815-7. No abstract available.
PMID: 13721076 [PubMed - OLDMEDLINE for Pre1966]

A brief history of Activator RNAs:

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euchromatin: "the most active portion of the genome within the cell nucleus".