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Lee B.-C.,Pharmacopuncture Medical Research Institute | Lee B.-C.,Korea Advanced Institute of Science and Technology | Kim H.B.,Seoul National University | Sung B.,Seoul National University | And 5 more authors.
Cardiology | Year: 2011

Background: Although there have been reports on threadlike structures inside the heart, they have received little attention. We aimed to develop a method for observing such structures and to reveal their ultrastructures. Methods:An in situ staining method, which uses a series of procedures of 0.2-0.4% trypan blue spraying and washing, was applied to observe threadlike structures on the surfaces of endocardia. The threadlike structures were isolated and observed by using confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM). Results: Networks of endocardial vessels (20 μm in thickness) with expansions (40-100 μm in diameter) were visualized; they were movable on the endocardium of the bovine atrium and ventricle. CLSM showed that (1) rod-shaped nuclei were aligned along the longitudinal direction of the endocardial vessel and (2) there were many cells inside the expansion. TEM on the endocardial vessel revealed that (1) there existed multiple lumens (1-7 μm in diameter) and (2) the extracellular matrices mostly consisted of collagen fibers, which were aligned along the longitudinal direction of the endocardial vessel or were locally organized in reticular structures. Conclusion: We investigated the endocardial circulatory system in bovine cardiac chambers and its ultrastructures, such as nucleic distributions, microlumens, and collagenous extracellular matrices. Copyright © 2011 S. Karger AG, Basel. Source


Novel threadlike structures (NTSs) on the surfaces of mammalian abdominal organs have recently attracted interests regarding their ability to transport fluid, enable cell migration, and possibly facilitate cancer metastasis. Nevertheless, histological studies of NTSs have been sporadic and often have inconsistent interpretations of the NTS internal structure. In this article, we provide a synthetic and consistent view of the NTS internal structure: the NTS is a loose bundle of fibrous stroma that forms interstitial channels and microsinusoids infiltrated with inflammatory cells. The fibroblasts are embedded in the stroma and mostly aligned along the major axis of the NTS. The sinusoids, which are in inconsecutive cross sections, have boundaries more or less delineated by extracellular fibers, partly surrounded by endothelial-like cells, or both. We compare these morphological features to other well-known connective tissues (i.e., trabecular meshwork and lymphatic capillary) and discuss the biomechanical and biological functions of NTSs based on their structural characteristics. Source


Lee B.-C.,Pharmacopuncture Medical Research Institute | Lee B.-C.,Korea Advanced Institute of Science and Technology | Lee H.-S.,Pharmacopuncture Medical Research Institute | Lee H.-S.,Sun Moon University | Kang D.-I.,Pharmacopuncture Medical Research Institute
JAMS Journal of Acupuncture and Meridian Studies | Year: 2012

According to Bonghan Kim's theory of anatomical reality for acupuncture meridians, DNA microgranules known as Sanals are key functional components in the primo vascular system (formerly the Bonghan system). To investigate this issue, we developed a new system, an incubator bound to a phase-contrast microscope, in which we cultivated and then observed for 10 hours microgranules taken from 3-day-old chick embryos and from blastoderms of fertilized chicken eggs. With this system, we found that, over time, the microgranules grew in circular patterns to become cell-like structures. In the embryo specimens, we found two distinctive microgranule growths, which developed into cell-like structures over 10 hours. In the first case, a microgranule of about 1.0 μm in size developed into a 3.3-μm-sized cell-like structure, with a pattern of concentric circles. The growth rate of the diameter of the first microgranule was, on average, 0.23 μm/hour. In the second case, a 2.5-μm-sized microgranule developed into a 5.4-μm-sized cell-like structure, which also exhibited a pattern of concentric circles. The average growth rate of the diameter of the second microgranule was 0.31 μm/hour. In the blastoderm specimens from the fertilized chicken egg, we also found three distinctive concentric growths. Interestingly, one of the three blastoderm microgranules grew very quickly, from about 2.5 μm in size to about 5.5 μm in size during 5 minutes of incubation. This was followed by steady growth to about 7.0 μm in size during the next 10 hours of incubation. In the final step of our investigation, we confirmed that the cell-like structures that had grown from the microgranules stained by acridine orange had DNA signals. We believe that the data obtained with our experimental method provide a clue that a mitosis-free alternative pathway for cell formation may, indeed, exist. We also suggest that this new function of microgranules (Sanals) might be related with the acupuncture meridian called the primo vascular system. © 2012. Source


Lee B.-C.,Korea Advanced Institute of Science and Technology | Lee B.-C.,Pharmacopuncture Medical Research Institute | Lee H.-S.,Korea Advanced Institute of Science and Technology | Lee H.-S.,Pharmacopuncture Medical Research Institute | And 2 more authors.
Micron | Year: 2013

With a combination of cultivation and phase-contrast and fluorescence microscopic observation, we first found that fusion of extracellular vesicles with or without membranes occurred in fertilized chicken eggs. In order to find solid evidence for fusion, we collected many fusion data from various tissues; primo vessels and pancreases of mice and pancreases and omentums of rats. Especially, by using acridine orange vital staining to demonstrate DNA and phase-contrast and fluorescence microscopy for long real-time observation, we found that many of the extracellular vesicles involved in the fusion process contained DNAs. The fusions fall into two main patterns: pattern A characterizes a fusion of less agitated extracellular vesicles without membranes. Pattern B is a fusion of vigorously vibrating extracellular vesicles in a certain membrane. Considering all data, tables, pictures and movies, we were able to show fusions of DNA extracellular vesicles without or with membranes in several tissues of three species. Interestingly, some of the fused structures share the same morphology as normal cell's in terms of overall shape, size and DNA signals in the center. Thus, in this article we first report the evidence for the fusion of extracellular vesicles with/without DNA toward a specific structure and discuss our findings by comparing with those of other pioneer's works in search for a mitosis-free alternative pathway for generating new cells. © 2012 Elsevier Ltd. Source


Lee H.-S.,Korea Advanced Institute of Science and Technology | Lee H.-S.,Pharmacopuncture Medical Research Institute | Lee B.-C.,Korea Advanced Institute of Science and Technology | Lee B.-C.,Pharmacopuncture Medical Research Institute | Kang D.-I.,Pharmacopuncture Medical Research Institute
Micron | Year: 2013

We found evidence that spontaneous self-assembly of DNA molecules from yolk granules occurred during the very early stage of egg fertilization. In order to find solid evidence for self-assembly of DNA molecules, we collected many available data in different stages of fertilized eggs, making a data table.At first by using acridine orange vital staining to demonstrate DNA, we noticed that some yolk granules emitted DNA signals that gradually increased with increasing incubation time from very small sizes to much larger nucleus-like structures. For convincing evidence, we also used another vital dye, Hoechst 33258 DNA-specific dye, to trace the changes in the yolk granules. The patterns of the DNA signals from yolk granules stained with Hoechst 33258 were the same as those from the yolk granules stained with acridine orange. A partial phase contrast microscopic image of the changes in the yolk granules showed some liquid-like material around the granules before the formation of the nucleus-like structures. Concomitant use of fluorescence and partial phase contrast microscopy suggested that these liquid-like materials may have been released from yolk granules in which spontaneous self-assembly of DNA molecules had occurred. Finally, in order to verify whether the DNA signals came from real DNA molecules or not, by using deoxyribonuclease I (DNAse), we confirmed that the nucleus-like structures were really assembled DNA molecules.Thus, in this article, we report evidence for the self-assembly of DNA molecules toward cell-like structures and discuss our findings, comparing them with those in the works of other pioneers, especially Antoine Béchamp, Olga Lepeshinskaya and Bong Han Kim, who insisted on the existence of a mitosis-free alternative pathway for generating new cells. © 2013 The Authors. Source

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