Sequential development of embryoblast like memory entities in human malignant tumors , an evidence of cancer cell metamorphosis retrodifferentiation.

Every cancer cell can partially or completely return to an embryonic genotype-phenotype: We capture the cycle of the activation of an individual cellular memory, which allowed a group of squamous tumor cells with mutations caused by the Human Papilloma Virus to return collectively to an embryoblast- like entities . Somatic injured cells have the plasticity to transform their morphology into an embryonic phenotype by expression of dormant genes when they enter a state of cellular emergency The hallmark of these entities is that its structure can exist in two visible chiral conformations and therefore behaves at distance molecular communication which results in clusters of malignant cells aligned in progressive order sequences, cancer retrodifferentiation recapitulate early stages of the embryogenesis with high immunopositivity expression for neuron specific enolase . Our findings document how malignant tissues reactivated ancestral storage memory and elaborate inside tumor glands crystalline proteins (Tc) biomimicry blastocyst- embryoblast fluid-filled cavity. Under physical cellular stress it is possible replicate these entities from the epithelium of minor salivary glands in humans determining that dormant genes expression can be induced by artificial stimulation. These observations confirm the importance of the foetal characteristics of cancer as a source of useful information that can contribute to better understanding of the biology of cancer at molecular, cellular and micro-environmental micro-environmental levels , to develop new target therapeutic alternatives not only in cancer but also in treatment of autoimmune, viral diseases, in regenerative medicine and rejuvenation. Initial experiments our laboratory show that we can artificially reproduce the assembly of these embryoblast-like entities by inducing slight controlled physical-chemical stress on the epithelium of minor salivary glands in healthy young individuals, as shown in the images . Expression of dormant genes can be induced by artificial stimulation.

identified in cancer tissues 10 years ago: interfacial geometry dictates cancer cell tumorigenicity [6], and matrix geometry determines the optimal cancer cell migration strategy and modulates responses to interventions. Whereas tumor cells exploit geometry for metastasis [7], the geometry helps confined cells to acquire a stem cell phenotype [8]. Today we postulate in accordance with our findings that this geometry of spirals and triangular patterns that we call triplet crystals (Tc) represents the crystalloid mold proteins on which the spatial order that gives rise to what we call embryoblast memory entities in malignant tumors is built.
Tumorigenesis resembles the self-organizing process of early embryo development. With the recent profound advances in the field of developmental biology, it has become apparent that the early development of embryos shares many similarities with cancer development in terms of both biological behaviors and similarities in the cell invasive epigenetic regulation of gene expression and protein profiling. Thus, it is evident that tumorigenesis mimics a self-organizing process of early embryo development [9][10][11][12][13][14][15]. The aim of this research is to demonstrate the intimate connection that cancer has with embryogenesis. Cancer recapitulate early stage of embriogenesis

Material and methods
We collected and re-examined all of our materials, in which we identified recurrent patterns of triangular and spiral chiral crystals (Tc) as geometric attractors, in cancer tissues. In the past 5 years, corresponding to more than 1077 microscopic/macroscopic specimens, including carcinomas, adenocarcinomas and sarcomas.
It is important to mention that the images documented in this work belong to living patients, with a natural history of cancer, who have not previously undergone chemotherapy or radiotherapy. The size of the giant masses documented show that they developed in approximately 9 years, with silent clinical growth. When these large masses were resected, none of these patients had developed metastases and it was for this specific reason that they were given the option of surgery as their condition allowed it.

Neuron-specific enolase -immunostaining
This isoenzyme, a homodimer, is found in mature neurons and cells of neuronal origin. Detection of NSE with antibodies can be used to identify neuronal cells and cells with neuroendocrine differentiation Sixty formalin-fixed and paraffin-embedded tissue sections with the most representative hot spot of Tc identified in malignant tumors were analyzed using neuron-specific enolase . We performed immunohistochemistry using the standard protocol method with paraffin sections. The scoring was done as follows: Ni (no immunostaining); low (10% or less immunopositivity); or high (>10% immunoreactive cells).

Statistics analysis
The index of Tc geometric complex assembly in cancer tissues was determined, as well as neuronspecific enolase antibody immunostaining positivity index in correlation with Tc expression areas.
Chi-squares for proportions were estimated using EPI-INFO software (v 6.04; Center for Disease Control and Prevention, Atlanta, GA, USA).
This study was approved by the ethics subcommittee of the University Cooperative of Colombia, Villavicencio, Colombia, and followed the guidelines of the Ministry of Health (No. 8430 of 1993) and the principles established by the Declaration of Helsinki. All patients signed an informed consent form for the use of their biological materials for diagnostic and research purposes.

RESULTS
From 1077 malignant tumors, Tc geometric complexes were identified in 1050 cases. These findings show identification of highly ordered geometric structures in more than 97.5% of the analyzed malignant tumor tissues (P = 0.00001; Table 1). Benign tumors and inflammatory entities do not evidence these structures. Tc represents the geometric triangular -spiral cleavage precursor in the formation of embryoblast entities.

Photomicrography evidence
We were able to capture a unique and perhaps unrepeatable image: the evolutionary cycle of structures with embryoblast-like phenotype generated from squamous epithelial cells injured by Human Papilloma Virus in a cervical cytology sample with cancer in situ. The cycle consists of 12 stages that show step by step how these structures are gestated.
As can be seen in the images, these entities are gestated from fractal crystalloid protein memory modules that have an embryoblast like phenotype, where clusters of malignant cells aligned in progressive order sequences with architectural metamorphosis from triangular -spiral cleavage microscopic -macroscopic structures, is activated when the cell suffers irreversible damage from specific mutations, as in this case through the action of the Human Papilloma Virus (Fig 1).
We identified three memory modules in this metamorphosis retrodiferentiation of these entities: a) Protein Crystalloid memory module: Stages 1 to 6 (Fig 1).
We can observe the intelligent sequence of a chain of events. Protein Crystalloid entities measuring 4 to 9 um appear separate, aligned and functionally interconnected. In stage 1 we can observe a polar entity made up of 2 molecular crystals proteins of triangular and spiral shapes that appear in a chiral position. We had previously described this event as GTCHC complexes [3] With this evidence, we can now state that these complexes initiate the process and originate from molecular liquids generated by the secretion of glandular tumor epithelia. These liquids inside the tumor glands generate movements for and against the clock, creating real molecular whirlwinds from which the spirals and triangles are formed, and when they solidify, they represent the crystalloid proteins of this process, which is in essence an eminently physical phenomenon. We wish to draw attention to the images documented below that show how malignant tumor glands adopt an emerging embryoblast -blastocyst like function. Remember that the blastocyst is an embryonic structure that forms during the early stages of development in which the morula develops as a fluid-filled cavity, transforming itself into a blastocyst or embryoblast.
In stages 2 to 5 we can see fractal copies of these protein crystalloid entities that have spontaneously replicated.
In stage 6, the entities gather all the information in a single structure and perforations can be seen on its surface. This entity full of holes on its surface and which we call Triplet Crystal (Tc) is constituted by a triplet of spirals and triangles perfectly and beautifully assembled spatially, which behaves as a mold-vector of biological information.

b) Cellular memory module
In stages 7 to 10 we can see the biological phase, which can be identified as the Tc pattern of stage 6 It behaves as a vector of biological information and transfers information to the nearby injured squamous cells, generating a change in these cells' normal polygonal cellular phenotype, sequentially acquiring the same pattern of the triangular phenotype of the Tc.
In stages 11 and 12 we can see the complete transfer of information from Tc to squamous cells, generating an exact copy of the Tc phenotype.
We can see how the transformed squamous cells progressively become more hyperchromatic as they reach stage 12, where an embryioblast like entity is clearly shown, completing the metamorphosis experienced by the squamous cells, probably regulated by the reactivation of genes and proteins that come from normal embryogenesis.
This evolutionary cycle shows other surprising details that speak for themselves: In the transition phase between stage 10 and 11, four pairs of triangular molecular satellite crystals can be seen near the squamous cells that are transforming their phenotype. These are once again perfectly visible in a chiral position (highlighted in a purple circle).
If we join the individual protein crystalloid memory modules with the spatially separated cellular memory modules, from stages 1 to 12, we obtain the spatial image of a perfect collective memory that has the phenotype of each of the individual memory entities. It can be observed how ""Every cellular collective" is identical to each of its individual cellular parts and each individual cellular memory has the phenotype of a collective memory encoded. With the unique patterns identified in this evolutionary cycle, we generated a prototype algorithm that allowed us to perfectly trace these entities in other tumor scenarios, and this is how we were able to identify them in carcinomas, adenocarcinomas and sarcomas. We had the opportunity to document these patterns in other scenarios under similar conditions. The Tc images show how this entity is an interface structure.
In panel h, A shows the cellular biological phase of the entity, given by its round phenotype, while B shows protein crystals and a geometric triangular-spiral pattern constituting the physical phase of the entity. At present, there is no evidence of a structure produced naturally where this structural conjunction between physics and biology can be so clearly observed, which makes this structure a mold to be copied and used in bioengineering as an entity that generates order and unique biological organization.     Due to their differentiation, these entities resist cellular hypoxia, evade the immune system, they are probably chemotherapy and radio resistance, and paradoxically, the tumors in which these entities are identified have a better prognosis than those tumors in which they do not occur. It is logical to assume that these cells and tissues that make up this entity no longer retain tumorigenic In the cephalic pole, the temporal bone petromastoid and eye periorbital area is recognized.
Material was captured in the ascitic fluid in a patient with colon cancer. h. i) Illustrate NS high immunopositivity of embryoblast like entities . The hallmark of these entities is that its structure can exist in two visible chiral conformations and therefore behaves at distance molecular communication which results in clusters of malignant cells aligned in progressive order sequences (green arrows).

Neuron enolase immunostaining
Tc represents the geometric triangular -spiral cleavage precursor in the formation of embryoblast entities with high immunopositivity expression for neuron specific enolase (95.0%) (table 2)  This corresponds to the macroscopic phase of the evolution of these entities that we have documented. To do this, we will rely on 7 specific cases with macroscopic and microscopic histological documentation of these cases in this phase of growth.      These patterns of sequentiality and order show that in the midst of the biological chaos and mutations that cancer represents, there is a product in development that is "gestating" as a result of the reactivation of signals from genes and proteins that return from embryogenesis.
we are facing probably with an emergent evolutionary biological response, we postulated that these entities that we are documenting are not there randomly, they have a function with the capacity to repair modify and or regenerate damaged senescent and mutated cells.
The genesis of these entities is given by the fluid secretions of the tumor glands that spin in the opposite direction, and solidify forming crystals of spiral triangular geometry, representing the crystalloid structural protein of these entities, which measure just a few microns but acquire polarity, organize, fuse and generate a triplet of triangular images and spirals forming a conglomerate that we have called triplet crystal (Tc). Tc represents the geometric triangularspiral cleavage precursor in the formation of these embryoblast like entities. .
In articles such as Magnetization of the three-spin triangular Ising model, theoretical physicists have already used mathematical formulas to address the organizing power of these geometric conglomerates, that take shape now in cancer biology and become real in practice in these self-organizing entities that we have documented here for the first time. Tc is a new, unique structure that nature builds at the interface of physics and biology where Geometric spin systems hold promise for finding new phases of biological self-assembly. [16 17] The theory of physics of a magnetization model forming a perfect conglomerate of a three-spin, three-triangle triplet is shown in the figures of the paper. In detail, a slender crystal-like Tc is able to behave as a vector of biological information and transmit its phenotype to squamous cells that change their phenotype in response to this transmitted information. As we can clearly see in the This article is the consolidation of over 10 years of work [19. 20]. A recent publication in an animal model by researchers from the neuroscience department of the University of California support our findings in human tissues, as they found that adult neuronal cells in mice exposed to cell damage return to a state of embryonic transcriptional growth state [21]. These findings are of unquestionable crucial significance for our research: These authors identify the transcriptional traces of embryonic growth, our group documents the entity that generated these traces, which determines that the methodology we used and the data we collected are reliable to the point that they can be reproduced and predicted in other laboratories around the world.
The hypothesis linking cancer and cell damage to embryogenesis is not new, Cohnheim suggested in 1882 [22] that tumor cells were essentially "embryonic" in nature, being remnants of embryonic epithelial cells. In the early 1970s, Brinster [23] demonstrated that by injecting embryonic carcinoma cells into a mouse blastocyst, the mouse was able to regulate the cancer cells and their progeny to the point that they no longer behaved malignantly; rather, they participated in normal embryonic development that resulted in functional mice. This experiment was confirmed by Mintz and Illmensee [24] and Papaioannou [25]. Pierce [26,27], showed that this effect, specific for some types of tumor cells, is strongly position-dependent: the carcinoma cells placed between the pellucid zone and the trophytoderm (the perivitelline space) were not controlled, while the carcinoma cells injected into the blastocele lost their tumorigenicity immediately after differentiation. Clinical trials, conducted with zebrafish embryo extracts administered to patients with advanced cancer that did not respond to conventional treatments, significantly reduced the expression of oncofetal antigens (such as AFP) [28] and induced marked beneficial effects (induction of objective responses, improvement in state performance and significant increase in overall survival) [29][30][31] .
Additionally, our morphological findings fit perfectly with the new hypothesis by molecular biologist Jose A from the University of Maryland who states that DNA is only "the list of ingredients" and not the set of instructions used to build and maintain a living organism. These instructions are very complex and are stored inside each individual cell as a shape memory that "decides" how and to what extent to use the ingredients available in the DNA. "DNA cannot be seen as the 'blueprint' for life" [32 33].
According to Dr. Jose the fundamental aspects of anatomy are dictated by something outside of DNA and he proposes that non-coding instructions in DNA are actually contained in the architectural arrangement of molecules within cells and in the interactions between them. This arrangement (shape memory) is what is preserved and transmitted from one generation to the next.
The findings of our study agree with the observations of Jose et al from the perspective that the entities found have their own identity and are unique since they have the differential phenotype of the host where they were gestated. In addition, the presence of visible pores or perforations also reported in the basement membrane of mouse embryos [Kyprianou et al [34] is noteworthy, supporting the theory that the entities found by us present characteristics of real "microscopic" embryos.
The analysis of living systems from molecular to population scales has revealed how the storage and processing of information across multiple scales is a key attribute of life, in which order can arise through the spontaneous association of molecules in the living system and the formation of dynamic structures (Tc) that can store and retrieve information from collections of self-assembled and selforganized molecules. These different ways of changing entities, sensors and properties highlight the multi-scale nature of living systems and suggest the usefulness of different entity-sensor-property frameworks at different molecular-microscopic-macroscopic scales that cannot be explained solely from the perspective of DNA or genome analysis.

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Our findings document how malignant tissues reactivated ancestral storage memory and elaborate, crystalline proteins repaired copies of the damaged substrate tissue. The resultant embryoblast template probably guides and controls the regenerative pathway mechanism in human tissues as follows: 1) Modify and reprogram the phenotype of the tumor where these entities are generated.
2) Establish a reverse primordial microscopic mold to use the collective behavior of cellular building blocks to regenerate injured tissues. 3) Convert cancer cells to a normal phenotype by developmental patterning of active patterning cues. 4) Convert cancer cells to a normal phenotype by regeneration using the organizational level and scale properties of reverse genetic guidance. 5) Globally control mitotic activity and morphogenetic movements avoiding their spread and metastasis, determining a better life prognosis for patients who incubate these entities in their tumors compared to those who do not express them.
These images clearly express, as never before, how a geometrical ordering entity (Tc) can somehow physically regulate the micro-macro cellular environment where these entities are gestated. We can predict these structures based on the knowledge of microscopic forces of self-assembly. At present, there is no documented structure produced naturally, where this structural conjunction between physics and biology is so clear and perfect, which makes it a real platform to be copied and used in the assembly and design of new proteins and artificial biostructures.
. These observations confirm the importance of the foetal characteristics of cancer as a source of useful information that can contribute to better understanding of the biology of cancer at molecular, cellular and micro-environmental levels.
Finally we believe that these isolated collective cellular entities are ready to be removed from their tumor microenvironment, as a product that has reached its maturity and has completed a cycle and is waiting to be used to develop new target therapeutic alternatives not only in cancer but also in treatment of autoimmune, viral diseases, in regenerative medicine and rejuvenation.