1. Masayuki Okumura 1.2.3.4..Marcelo Mester 1, Carlos E. P. Corbett 2 & Joaquim Gama-Rodrigues 1 Department of Gastroenterology 1 and Pathology 2 of Faculdade de Medicina da Universidade de S. Paulo, Instituto de Medicina Tropical de S. Paulo 3 and Instituto Adolfo Lutz, S. Paulo 4, Brazil. Remember: Right is always the laboratory animal; it is irrational and does not know the “ blindman and elephant” tale. INTRODUCTION At the moment the scientists expose the lab animals to the X-ray, they are destroying cellular matrix. Due to the lack of cells, they admit the cellular dedifferentiation or “plasticity” 1, which consist of changing grown-up cells into primitive totipotent cells they are also mind embryos stem or trunk cells. “Stem or trunk cells” are part of a tree with pre-established genetic code and they should develop into specific fruits according to the seeds, without behavior of fertile grounds. HYPOTHESIS Our daily contact with two of the endemic tropical diseases of Brazil enabled us to propose the following hypothesis: a “Fruitful jelly or Amorphous primitive mass” is precursor of the “Stem or Trunk Cells” of modern scientists. During the second half of the last century (1958) we initiated: 1)the surgical TREATMENT OF PORTAL HYPERTENSION in patients with Hepatosplenic Schistosomiasis (Schistosoma mansoni) with one of the techniques employed: Splenectomy + Spleno-renal shunt”. The patients frequently presented changes of their hemograms, often presenting pancytopenia (anemia with RBC values of 2,500,000 – 3,000,000/mm 3 ; leucopenia with WBC values of 2,000 – 3,000/mm 3 and thrombocytopenia with platelet values of 20,000 – 40,000/mm 3 ). We noted that two to four days posoperatively, hypersplenism had already been corrected. Some patients even presented platelet levels of 500,000 to 1,000,000 units/mm 3. Our “Fruitful jelly’ or Amophous Primitive mass” remained at rest, undifferentiated until summoned by a splenectomy, they reacted roughly by liberating billion cells or leukemic reaction. A)If the parasite enters the bone marrow (Fig. 1) or spleen, it progresses freely in the shape of an eosinophilic ring which upon breaking, acquires the shape of a truncated “C” (Fig. 2) and an elongated “C”, seeming to develop by an “indirect mechanism or uncoiling” (Fig. 3) 5.- medullar figure 10. Krause 4 from Yale University School of Medicine estimates a ratio of “only one in 100,000 bone marrows cells may be stem cells” I would like to draw attention to Figs. (2, 7 and 8) which show the eosinophilic ring shaped and spheric of cells which we believe are the stem cells or trunk cells. In fact, they have been showed by other researchers (Chagas (1909) 3, Silva & Camargo (1968) 5, Pinto et al. (1999) 6, Kondo (2000) 7, and recently at A.A.A.S. – Annual Meeting and Science Innovation Exposition, held on 14-19 February 2002 – Boston MA, on Stem Cells Biology: From Basic Research to Clinical Opportunity, by Rossant 8 and Goldman 9. B)If entering the liver (Fig. 4), it develops a spherical shape with basophils particles forming a bulge (Fig. 5) in the membrane followed by the flagellum and undulating membrane, acquiring the shape of a tadpole (Fig.6) and evolving by a “direct mechanism or elongation 9 ”- hepatic figure. 564 The presence of ring-shaped figures in the liver was unusual. The ratio was hundred of hepatic forms to only one medullar (Fig. 7). The opposite occurred in the bone marrow (Fig. 8), leading us to suppose that the amorphous primitive mass remained at rest, quiescent state, undifferentiated, until summoned by an abrupt aggression (roughly) they reacted by liberating the two forms. We can presume that trunk or stem cells work like joined antibodies, therefore, they do not exist normally, but only after stimulation. 7 8 C)In any other cells, the parasite is transformed from a flagellate (tripomastigote – Fig. 18), into an elliptical one (amastigote) and during reproduction the descendants remain grouped forming a pseudocyst or parasitic nest 3 (Fig. 10 - 20). The parasite develop to epi and tripomastigote ( adult form); the increase volume causes the nest break (Fig. 11 – 12), releasing a cytokine-like substance 11 (Fig. 11- 12 - 14) (constitute by conjugation of a pro-cytokine originating from the parasite nest with its DNA, with another pro-cytokine of host cell lesion and with its genetic code) and tripomastigote. Trypanosoma cruzi (Fig. 18) began in the blood stream on the fourth day and increase teathing its maximum on the seventeenth day. It continued rising until the twenty second day, dropping quickly, attaining the minimum and disappearing from the circulation at the beginning of the second month. Parasitism characterized by the presence of amastigotes nests in the heart did not take place with the parasitic bloody elevation; the maximum was attained on the nineteenth day of illness. After the twenty-first day, it decreased abruptly parallel to the drop in blood stream, becoming negative after thirthieth day (Fig. 9). 123 Sternal bone marrow : amastigote nest ( histology, 400 X. H.E.) Sternal bone marrow, imprint: ring – shaped and evolutive forms resulting from unwinding. (Giemsa – photo Kodak film) Bone marrow, imprint: parasite evolved by an “unwinding or indirect mechanism” (Giemsa. - photo Kodak Film) Liver, histology: slide with free parasites in the capillaries and sinusoids (Giemsa. – photo Kodak Film) Liver, imprint: spherical and developing forms resulting for elongation (Giemsa. – photo Fuji Film) Liver, imprint: Evolutionary form resulting from elongation – resembling tadpole. (Giemsa. – photo Kodak Film) BACKGROUND 2)we began to study the correlation between EXPERIMENTAL CHAGA’S DISEASE and “megas” in laboratory animals. When researching experimental Chagas’disease 2 and inoculating Trypanosoma cruzi 3 in mice, we found that the protozoan enters the organism’s cells for its reproduction: Sternal bone marrow, imprint (medullar form):a) eosinophilic ring-shaped, b) C and spindle-shaped amastigotes forms. C) tadpole and d) three hepatic forms, free in medullar fat (Giemsa – photo Fuji Film). Liver, imprint (liver form): spheric cells, with increased volume containing basophilic particles and three medullar forms free in the interstitial space (Giemsa).

2. BRAZILIAN CLONING THEORY For the purpose of understanding the cloning, let us hypothesize that our matrix as a behaves similarly to the ovular cytoplasm. When scientists remove the ovulum’s nucleus to initiate cloning, they are transforming into our multitipotent “Fruitful Jelly or Amorphous Primitive Mass”, with the capability of making any organs, they put the cell to be cloned producing an embryonic stem cells or trunk cells THE DREAMS OF PIONEERS IN HUMAN ORGANS TRANSPLANTS Our opinion is that only molecular biology will be able to definitely clarify the participation of our “Fruitful jelly or Amorphous Primitive Mass”, in the genesis, repair and transformations tissues. This will certainly be an important step on the way to producing tissues “in vitro” and even CHANGING THE TRANSPLANTS TO REPAIR by the simple lesion (cut or biopsy) of target organs. b- If the reaction is inadequate, the cytokine-like substance entering the general circulation (pleiotropy), will stimulate the hematopoietic organs, activate our “FRUITFUL JELLY or AMORPHOUS PRIMITIVE MASS” (Fig. 16 - 17) and transform it into WBC’s (lymphocytes “activated” and macrophages (Fig. 19 – 20)). The last WBC are oriented by the original host cells genetic code, is directed to the respective organ, constituting the acute inflammatory process (Fig. 13). This coincide with the appearance of antibodies in the blood, achieving very high concentration. a- When forming in the interstitial space, the cytokine-like substance 11 by an autocrine or paracrine mechanism attracts the WBC’s (neutrophils, lymphocytes naïve and monocytes) from the local capillaries, forming the first organic reaction. c- When this defense mechanism would be activated with especial combination with cytokines, as an attempt to surround the lesion which may form incipient granuloma (Fig. 15), with macrophages, mastocytes, proliferation of fibroblasts (sub-acute) leading to fibrosis and residual scars (chronic reaction). 9 Gel or amorphous primitive mass. Blood stream: two adults of Trypanosoma cruzi. Macrophage filled with ring shaped of T. cruzi. Macrophage filled with amastigotes 1718192016 SUMMARY AND CONCLUSION We can admit that our “fruitful jelly or amorphous primitive mass” is the precursor of the trunk or embryonic stem cells, emphasizing: REFERENCES KRAUSE, D.S.- et al. – Multi-organ, multi lineage engraftment by a single bone marrow – derived stem cell. Cell, 2001, 105: 369-377. SILVA, L.H.P. & CAMARGO, E.P.- Ciclo evolutivo do Trypanosoma cruzi. In CANÇADO, J.R.- Doença de Chagas. Imprensa Oficial de Belo Horizonte, Minas Gerais, 1968, 87-99. PINTO, P.L.S. et al.- Life cycle of Trypanosoma cruzi ( Y strain) in mice. Rev. Hosp. Clin. Fac. Med. S. Paulo, 1999, 54: 141-146. RIBEIRO, R.B. et al.- Relato sobre a infecção crônica pelo Trypanosoma cruzi (cepa Y) em camundongos Swiss. Rev. Brasileira de Medicina Tropical, 1999, 32 sup. 1: 334. VIANNA, G.- Contribuição para o estudo da anatomia patolojica da “Moléstia de Carlos Chagas”. Mem. Inst. Oswaldo Cruz, 1911, 3: 276-294. OKUMURA, M. et al.- Cytokine – like substance: origin and fate in Chagas’ disease. New hypothesis about the local inflammatory reaction. Rev. Hosp. Clin. Fac. Med. S. Paulo, 1999, 54: 73-74. OKUMURA, M.- Doença de Chagas Experimental. In RAIA, A.A.- Manifestações Digestivas na Moléstia de Chagas. Sarvier, S. Paulo, 1983, pp. 35-59. NOTE: Trypanosoma cruzi was chosen because it is easily visible under the optical microscope due to its size, it has great mobility, functioning as a biological marker and even eliminating the use of stains DESCRIPTORS: Fertile ground, Stem Cells. Trunk Cells, Schistosoma mansoni, Trypanosoma cruzi, Hematopoiesis. Cytokine, Cloning 3)besides the daily variety in number and size. 2)its versatility for change of shape: flagellate – elongated (tripomastigote), elliptical (amastigote); round (RBC, WBC); cubic, paving and columnar (epithelial); spindle (connective tissue and muscle), star-shaped with its branches (neuron, nerve), calcified (bone), etc, and 1)its potential for furnishing the raw material for forming: a) the cell; b) the body of the parasites; c) the red, white blood cells and platelets and d) the energy; HOLDEN, C & VOGEL, G.- Plasticity: Time for A Reappraisal? Science, 2002, 296: 2126- 2129. OKUMURA, M. & CORRÊA NETTO, Produção Experimental de “MEGAS” em Animais Inoculados com Trypanosoma cruzi. Rev. Hosp. Clin. Fac. Med. S. Paulo, 1961, 16: 338- 341. CHAGAS, C.- Nova tripanosomiase humana. Estudo sobre a morfologia e o ciclo evolutivo do Schizotrypanum cruzi, n. gen. n. sp, agente etiológico de nova entidade mórbida do homem. Mem. Inst. Oswaldo Cruz, 1909, 2: 1-62. KONDO, M. et al.- Cell-fate conversion of lymphoid-commited progenitors by instructive actions of cytokines. Nature, 2000, 407: 383-386. ROSSANT, J.- Stem Cells in the Early Embryo. A.A.A.S.- Annual Meeting and Science Innovation Exposition, 14-19 February, Boston, MA, 2002: 56. GOLDMAN, S.A.- Neural Stem and Progenitor Cells of the Human Brain. A A A S.- Annual Meeting and Science Innovation Exposition, 14-19 February, Boston, MA, 2002: 56. D) Because of the cyclical nature of the disease, its repetition determines the increase in volume of the target organs, originating the respective “megas” (megaesophagus, megastomach, megaduodenum, megacolon, cardiomegaly, in addition to hepatosplenomegaly) 13 (Fig. 21 – 22 – 23 - 24). Graphic showing parasitemia and parasitism. 11 10 12 13 14 Colon, Auerbach plexus: integral neuron with parasites without inflammatory reaction (histology- HE. 400X) Colon, Auerbach plexus rupture of nest within parasitic neuron and ruptured of host cell without inflammatory reaction (cytokine should be forming at this time) (histology. HE 400 X) Colon, Auerbach plexus: rupture neuron without reaction ( histology- HE. 400X). 13- Colon, Auerbach plexus: Begnning of acute inflammatory process. Colon, Auerbach plexus: Lymphocytes stained cytokine. 15 Colon, Auerbach plexus: Granuloma formation 23 Hepatosplenomegaly and mega bladder 21 Megastomach, Megaduodenum and Rx. Animal preserved in formalin. 22 Evolutive forms of Chagasic cardiomegaly 24 Megaesophagus, megastomach, megaduodenum and normal control: X rays with contrast: meagesophagus with achalasia of the cardia and megacolon with achalasia of pelvi-rectal sphincter.