• Biomolecules & Therapeutics
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• v.14 no.2
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• pp.90-95
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• 2006

CKD-602 exerts its antitumor effect via inhibition of topoisomerase I in cancer cells. Multicellular spheroid (MCS) and Multicellular layers (MCLs) are known as in vitro 3-dimensional models which closely represent tumor conditions in vivo. In order to investigate the potential of CKD-602 against human colorectal tumors, we evaluated the anti-proliferative activity and penetration ability of CKD-602 in MCS and MCL cultures of DLD-l human colorectal cancer cells, respectively. The maximum effects($E_{max}$) induced by CKD-602 were significantly lower in MCS compared to monolayers (48% vs 92%). With prolonged drug exposure, the $IC_{50's}$ of CKD-602 decreased to $23.5{\pm}1.0nM$ in monolayers after 24 h exposure and $42.3{\pm}1.7nM$ in MCS after 6 days, respectively. However, no further increase in effect was observed for exposure time longer than growth doubling time (Td) in both cultures. Activity of CKD-602 was significantly reduced after penetration through MCL and also with cell-free insert membrane. In conclusion, CKD-602 showed significantly decreased anti-proliferative activity in 3D cultures (MCS) of human colorectal cancer cells. Tumor penetration of CKD-602 could not be determined due to loss of activity after penetration through cell free insert membrane, which warrants further evaluation using a modified model.

• Korean Journal of Ichthyology
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• v.19 no.3
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• pp.201-209
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• 2007

The fine structures and histochemical features on the integumentary system of the grass puffer, Takifugu niphobles were examined by means of the light and transmission electron microscopy. Integumentary surface of the grass puffer showed irregular folds in light microscope. The folds of the ventral region are more pronounced than those in the dorsal region. Integumentary system is composed of outer epidermal layer and inner dermal layer. The stratified epidermal layer consists of epithelia, mucous cells, club cells, granular cells and multivacuolar gland. Epithelial cells are classified into superficial, intermediated and basal cell, and free surface of superficial cell is covered with microridges. Glands of the epidermal layer are divided into unicellular and multicellular gland. Mucous cells of multicellular gland contains mucosal materials of neutral glycoprotein. Multivacuolar gland is composed of numerous vacuole cells of about $20{\mu}m$ in axial diameter. Vacuole cells contains a large central vacuole and are connected to another by many desmosomes. The mucous glands and multivacuolar glands are more abundant in ventral region than dorsal integument. The thickness of dermis is more three to five times than epidermis in ventral integument. The collagen fibers, fibrocytes, nerve cells, basal plate of spine and chromatophore are observed in the dermal layer of compact connective tissue.

• Animal cells and systems
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• v.12 no.2
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• pp.85-91
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• 2008

Structure and mucosubstance histochemistry in the epidermis of five teleostean species, i. e., spotty belly, Agrammus agrammus, devil stinger, Inimicus japonicus, stone fish, Erosa erosa, cubed snailfish, Liparis tessellatus, and Japanese bluefish, Scombrops boops were investigated. The epidermis of five species studied is composed of three layers: superficial, middle, and basal layer. The superficial layer is comprised of rather flattened cells. Mucous cells, the type commonly found in fishes are completely lacking in the epidermis of devil stinger, stone fish, and cubed snailfish. The epidermis of devil stinger and stone fish have multicellular glands which do not have mucosubstances. The skin surface of them is covered with mucous layer. The superficial cells in the epidermis of devil stinger, stone fish, and cubed snailfish are mucus-secreting cells. The composition and the amount of the mucosubstances vary in species and body regions. The mucous layer on the skin surface and superficial epidermal cells of devil stinger contain a mixture of neutral and acidic(sulfated and nonsulfated) mucosubstances. In stone fish, the mucous layer has acidic(sulfated and non-sulfated) mucin and the superficial epidermal cells contain neutral mucin. In cubed snailfish, the type of epidermal mucosubstances is identified as a mixture of neutral and acidic(non-sulfated) mucin. The mucous cells of the epidermis in spotty belly and Japanese bluefish contain neutral mucin.

• Journal of Life Science
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• v.10 no.1
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• pp.37-44
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• 2000

The mammary gland contains a subpopulation of epithelial cells with large proliferative potentials which are the likely targets for carcinogens. These clonogenic cells can proliferate and differentiate into functional glandular structures. Rat mammary epithelial cells (RMEC) were isolated and characterized in vitro. By flow cytometry of RMEC stained with fluorescein isothiocyanate-peanut agglutinin(PNA) and phycoerythrin anti-Thy-1.1 monoclonal antibody, it was possible to four cell subpopulations from 7-8 week old F344 female rat mammary glands: cells negative to both reagents (B-), PNA-positive cells (PNA+), Thy-1.1-positive cells (Thy-1.1+), and cells positive to both reagents (B+). When single PNA+ cells were isolated and cultured in Matrigel with irradiated (∼50 Gray) 3T3 fibroblast feeder layer, they gave rise to multicellular clonal structures of three types: alveolar, foamy alveolar, and squamous colonies. The developed structures were similar to the mammary glands in vivo. These results suggest that some of PNA+ cells possesses many of the characteristics of multipotent clonogenic stem-like cells.

• Applied Microscopy
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• v.52
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• pp.4.1-4.11
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• 2022

Field emission scanning electron microscopy (FESEM) is an essential tool for observing surface details of specimens in a high vacuum. A series of specimen procedures precludes the observations of living organisms, resulting in artifacts. To overcome these problems, Takahiko Hariyama and his colleagues proposed the concept of the "nanosuit" later referred to as "NanoSuit", describing a thin polymer layer placed on organisms to protect them in a high vacuum in 2013. The NanoSuit is formed rapidly by (i) electron beam irradiation, (ii) plasma irradiation, (iii) Tween 20 solution immersion, and (iv) surface shield enhancer (SSE) solution immersion. Without chemical fixation and metal coating, the NanoSuit-formed specimens allowed structural preservation and accurate element detection of insulating, wet specimens at high spatial resolution. NanoSuit-formed larvae were able to resume normal growth following FESEM observation. The method has been employed to observe unfixed and uncoated bacteria, multicellular organisms, and paraffin sections. These results suggest that the NanoSuit can be applied to prolong life in vacuo and overcome the limit of dead imaging of electron microscopy.

• Applied Microscopy
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• v.51
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• pp.16.1-16.7
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• 2021

Electron microscopy (EM) is an essential imaging method in biological sciences. Since biological specimens are exposed to radiation and vacuum conditions during EM observations, they die due to chemical bond breakage and desiccation. However, some organisms belonging to the taxa of bacteria, fungi, plants, and animals (including beetles, ticks, and tardigrades) have been reported to survive hostile scanning EM (SEM) conditions since the onset of EM. The surviving organisms were observed (i) without chemical fixation, (ii) after mounting to a precooled cold stage, (iii) using cryo-SEM, or (iv) after coating with a thin polymer layer, respectively. Combined use of these techniques may provide a better condition for preservation and live imaging of multicellular organisms for a long time beyond live-cell EM.

• Applied Microscopy
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• v.30 no.3
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• pp.311-319
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• 2000

The gill morphology and ultrastructure of the fleshy shrimp, Penaeus chinensis were investigated by light and electron microscopy. Fleshy shrimp has dendrobranchiate gills. Gill has a longitudinal septum dividing them into afferent and efferent channel. Each gill lamella is covered by multi-layered thin cuticle of different electron density. The lamella basal cell is squamous and contains cytoplasm of electron dense. Simple epithelial layer consists of squamous epithelium contained large nucleus. The lamella pillar structures are characterized by the axial microtubules and lateral membrane interdigitations Secretory cells of AB-PAS negative are multicellular gland. In active gland each cell boundary is not apparent and the cytoplasm contains smooth endoplasmic reticula, mitochondria, membrane-bounded secretory vesicles of low electron density and granular resettes. In inactive gland each cell boundary is apparent and the cytoplasm is occupied with numerous small granules of electron dense. The well-developed rough endoplasmic reticula and Golgi apparatus are observed in the unicellular gland of alcian blue positive.

• Journal of computational fluids engineering
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• v.21 no.4
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• pp.11-18
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• 2016

A numerical study is conducted on the optical fiber coating flow in a primary coating nozzle consisting of three major parts: a resin chamber, a contraction and a coating die of small diameter. The flow is driven by the optical fiber penetrating the center of the nozzle at a high speed. The axisymmetric two-dimensional flow and heat transfer induced by viscous heating are examined based on the laminar flow assumption. Numerical experiments are performed with varying the convergent angle of nozzle contraction and the optical fiber drawing speed. The numerical results show that for high drawing speed greater than 30 m/s, there is a transition in the essential flow features depending on the convergent angle. For a large convergent angle greater than $30^{\circ}$, unfavorable multicellular flow structures are monitored, which could be associated with wall boundary-layer separation. In the regime of small convergent angle, as the angle increases, the highest resin temperature at the exit of die and the coating thickness decrease but the sensitivity of coating thickness on drawing speed and the maximum shear strain of resin on the optical fiber increase. The effects of the convergent angle are discussed in view of compromise searching for an appropriate angle for high-speed optical fiber coating.

• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.3
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• pp.470-486
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• 1995

Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.

• Applied Microscopy
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• v.38 no.1
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• pp.21-28
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• 2008

Carnivorous plants vary in their unique features of morphology, ultrastructure and biochemical properties by species. Furthermore, prey-capturing mechanism as well as structural and physiological adaptations have been used for grouping various carnivorous species. In Drosera plants, glandular trichomes, which develop in the leaf epidermis, are known to play the most important role during the prey capturing process. The present study examined such trichomes, focusing on the glandular type, in leaves of Drosera anglica using scanning and transmission electron microscopy. Three types of rudimentary glandular trichomes were found to develop within the folded leaf primordia and immature leaf during early development. The first type, stalked glandular trichomes (Type I), occurred on the margin and upper epidermis of the leaf. With maturation, the longest glandular trichomes having lengthy stalks, ca. $2.2{\sim}5.1\;mm$, developed along the margin, while shorter stalked trichomes, ca. up to $200\;{\mu}m$, were found on the inner leaf blade. The shorter ones consisted of a globose head having two layers of secretory cells, parenchyma bell cells and tracheids and a multicellular stalk. The stalks gradually decreased in length in centripetal fashion. The second type, Type II, having ca. $15{\sim}30\;{\mu}m$ short stalks, also developed along the inner blade. Both types secreted mucilage from the secretory cells which had a thin cell wall and cuticle layer. The sessile six-celled glandular trichomes were the third type, Type III, and were $25{\sim}40\;{\mu}m$ in length. They were distributed most commonly throughout the upper and lower epidermis, petiole and even on the stalk surfaces of the first two types of trichomes. The third type was also found to be involved in the active secretion. In prey capturing leaves, all trichome types secreted substances through thin cuticles in the head cell wall, which exhibited relatively loose wall components.