• KSBB Journal
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• v.19 no.2
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• pp.118-124
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• 2004

Melanocytes synthesize melanin within discrete organelle termed melanosomes which are transferred to the surrounding keratinocytes and can be produced in varying sizes, numbers and densities. Skin whitening products have become increasingly popular in the past few years. The most successful natural skin whitening agents are: Arbutin, Vitamin C, Kojic acid, Mulberry, which are all tyrosinase inhibitors. In this work, melanoston, a melanogenesis inhibitor isolated from yeast was studied to understand its mechanism of melanogenesis inhibition. It was found that melanoston was not a tyrosinase inhibitor, while when melanoston was applied to the B16 melanoma cell culture media, the intracellular tyrosinase activity was decreased by more than 30％, When B16 melanoma was stimulated with ${\alpha}$-MSH, cell morphololgy was dramatically changed to have lots of dendrites on the cell membrane surface. On the other hand, B16 was treated with ${\alpha}$-MSH and melanoston, simultaneously, the change of cell morphology was not so great. This inhibition effect of melanoston was found to be related to the inhibition of intracellular activation and transportation of tyrosinase, which was observed by immunostaining of B16 melanoma using anti-tyrosinase antibody. From these results, melanoston was regarded as an inhibitor to the differentiation of melanoma cells.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.1 s.49
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• pp.25-33
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• 2005

Melanocytes synthesize melanin within discrete organelle termed melanosomes which are transferred to the surrounding keratinocytes and can be produced in varying sizes, numbers and densities. Skin whitening products have become increasingly popular in the past few years. The most successful natural skin whitening agents are: arbutin, vitamin C, kojic acid, and mulberry, which are all tyrosinase inhibitors. In this work, melanoston, a melanogenesis inhibitor isolated from yeast was studied to understand its mechanism of melanogenesis inhibition. It was found that melanoston was not a tyrosinase inhibitor, while when melanoston was applied to the B16 melanoma cell culture media, the intracellular tyrosinase activity was decreased by more than $30\%$. When B16 melanoma was stimulated with $\alpha$-MSH, cell morphololgy was dramatically changed to have lots of dendrites on the cell membrane surface. On the other hand, B16 was treated with $\alpha$-MSH and melanoston, simultaneously, the change of cell morphologv was not so great. This inhibitory effect of melanoston was found to be related to the inhibition of intracellar activation and transportation of tyrosinase, which was observed by irmmunostaining of B16 melanoma using anti-tyrosinase antibody. From these results, melanoston was regarded as an inhibitor to the differentiation of melanoma cells.

• Journal of Microbiology and Biotechnology
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• v.25 no.5
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• pp.718-722
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• 2015

Techniques for immobilizing effective enzymes on nanoparticles for stabilization of the activity of free enzymes have been developing as a pharmaceutical field. In this study, we examined the effect of three different pH conditions of phosphate buffer, as a dissolving solvent for lysosomal enzymes, on the direct immobilization of lysosomal enzymes extracted from Hen's egg white and Saccharomyces cerevisiae. Titanium(IV) oxide (TiO₂) nanoparticles, which are extensively used in many research fields, were used in this study. The lysosomal enzymes immobilized on TiO₂ under each pH condition were evaluated to maintain the specific activity of lysosomal enzymes, so that we can determine the degree of melanin treatment in lysosomal enzymes immobilized on TiO₂. We found that the immobilization efficiency and melanin treatment activity in both lysosomal enzymes extracted from Hen's egg white and S. cerevisiae were the highest in an acidic condition of phosphate buffer (pH 4). However, the immobilization efficiency and melanin treatment activity were inversely proportional to the increase in pH under alkaline conditions. In addition, enhanced immobilization efficiency was shown in TiO₂ pretreated with a divalent, positively charged ion, Ca²⁺, and the melanin treatment activity of immobilized lysosomal enzymes on TiO₂ pretreated with Ca²⁺ was also increased. Therefore, this result suggests that the immobilization efficiency and melanin treatment activity of lysosomal enzymes can be enhanced according to the pH conditions of the dissolving solvent.

• Development and Reproduction
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• v.11 no.2
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• pp.79-84
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• 2007

The freezing susceptibilities of two larval stages (D-shaped and umbo) of the pearl oyster (Pinctada fucata martensii) were evaluated by the electron microscopy (light, transmission electron and scanning electron). The morphological shapes were examined from each pre-frozen or frozen-thawed stage of the cryopreserved larvae in liquid nitrogen by using the cryoprotectant, dimethyl sulfoxide ($Me_2SO$) mixed with sucrose. Although a portion of the shell was damaged, the hinge and prodissoconch were intact and clearly visible after preservation in liquid nitrogen. In addition, the cytoplasm of the frozen-thawed larvae maintained the normal organelle integrities, e.g., endoplasmic reticula, lipid droplets, mitochondria, nucleus and microvilli. However, some of the frozen-thawed larvae showed irregularly arranged cilia, rough shell surfaces and round-lumped cilium heads. These results indicate that P. fucata martensii larvae are susceptible to freezing, at least at those two critical developmental stages (D-shaped and umbo), and suggest a new industrial investigation including reduction method of cell injury for preserving microbial starter cultures need to be developed.

• Molecules and Cells
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• v.40 no.6
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• pp.401-409
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• 2017

The primary cilium is a non-motile microtubule-based organelle that protrudes from the surface of most human cells and works as a cellular antenna to accept extracellular signals. Primary cilia assemble from the basal body during the resting stage ($G_0$ phase) and simultaneously disassemble with cell cycle re-entry. Defective control of assembly or disassembly causes diverse human diseases including ciliopathy and cancer. To identify the effective compounds for studying primary cilium disassembly, we have screened 297 natural compounds and identified 18 and 17 primary cilium assembly and disassembly inhibitors, respectively. Among them, the application of KY-0120, identified as Brefeldin A, disturbed Dvl2-Plk1-mediated cilium disassembly via repression of the interaction of $CK1{\varepsilon}-Dvl2$ and the expression of Plk1 mRNA. Therefore, our study may suggest useful compounds for studying the cellular mechanism of primary cilium disassembly to prevent ciliopathy and cancer.

• Animal cells and systems
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• v.16 no.2
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• pp.104-113
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• 2012

The nucleolus is a distinct nuclear territory involved in the compartmentalization of nuclear functions. There is some evidence of a relationship between nuclear fragmentation during spermatogenesis and chromatoid body (CB) formation. The CB is a typical cytoplasmic organelle of haploid germ cells, and is involved in RNA and protein accumulation for later germ-cell differentiation. The goal of this study was to qualitatively and quantitatively describe the nucleolar cycle during the spermatogenesis of $Phrynops$ $geoffroanus$ (Reptilia Testudines), and compare this nucleolar fragmentation with CB formation in this species through the use of cytochemical and ultrastructural analysis. Qualitative analysis showed a fragmentation of the nuclear material after pachytene of the first meiotic division in the primary spermatocytes. Quantitative analysis of the nucleolar cycle revealed a significant difference in the number of nucleoli and in the size of the nucleolus between spermatogonia and early spermatids. Using ultrastructural analysis, we recorded the beginning of the CB formation process in the cytoplasm of primary spermatocytes at the same time as when nuclear fragmentation occurs. In the cytoplasm of primary spermatocytes, the CB was observed in association with mitochondrial aggregates and the Golgi complex. In the cytoplasm of early spermatids, the CB was observed in association with lipid droplets. In conclusion, our data show that the nucleolus plays a role in the CB formation process. During spermatogenesis of $P.$ $geoffroanus$, the CB is involved in some important biological processes, including acrosome formation and mitochondrial migration to the spermatozoon tail and middle piece region.

• Applied Microscopy
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• v.33 no.3
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• pp.233-241
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• 2003

Patterns of tannin accumulation in leaves of C-4 Euphorbia maculata have been examined using electron microscopy. Tannins, which are secondary metabolite phenolic compounds, were found to be deposited conspicuously in vacuoles of certain tissues regardless of their stage in development. However, patterns of deposit accumulation were distinguishable by their cell type during leaf differentiation. The deposits appeared most concentrated in the concentric bundle sheath cells enclosing veins, while little or no density was detected mostly in the mesophyll cells close to the epidermis. An ultrastructural study revealed that the deposits were restricted to the vacuoles at an early stage of leaf development; during which the vacuoles were almost completely filled with the tanniferous substances. The deposits themselves took different forms ranging from granules to huge globules while expanding leaf blade. As the leaf matured, the deposits accumulated either centripetally adjacent to the inner tangential tonoplast or by penetration into the cytoplasm amongst various cellular organelles, resulting in an extremely dense cytoplasm. Electron micrographs frequently showed the delineation of each organelle by the presence of dense deposits within the cytoplasm. Some large depository vacuoles filled with tannins had a corrugated appearance on the sectioned surface. The pattern and potential role of the deposits have been discussed.

• Applied Microscopy
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• v.27 no.4
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• pp.433-452
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• 1997

Mongolian gerbil (Meriones unguiculatus) has been as an animal model for studing the neurological diseases such as stroke and epilepsy because of the congenital incompleteries in Willis circle, as well as the investigation of water metabolism because of the long time-survival in the condition of water-deprived desert condition, compared with other species animals. In order to accomplish the this research, first of all another divided the laboratory animals 5 groups of which each group include the 5 animals. In this study were investigated the histological structure in the kidney, measured the plasma osmolalities at the time of sacrifice of indivisual animals, and the body weights every day during water-deprived. The results obtained in this study were summarized as followings: 1. The body weights and decreasing rates of the body weight in water-deprived mongolian gerbil groups were continuosly decreased. 2. The plasma osmolalities were increased from the 5th water-deprived day, after then the gradually increasing reached nearly its equilibrium state at the 10th water-deprived day. 3. The urine volumes were abruptly decreased from the 2th water-deprived day, after then the gradually decreasing patterns were reached nearly its equilibrium state at the 10th day, and stopped the 11th day. 4. In the light microscopical observation of the kidney, glomerular capillary loop thickening, mesangial matrix increasing, sclerosis, glomerular cystic atrophy, interstitial fibrosis, tubular dilatation, mononuclear interstitial inflammation, interstitial mineralization, and hyperplasia of the collecting duct epithelium in the cortex area, were observed from the 10th water deprived day, and the lesions were gradually severe changed as the time lapse. 5. In the electron microscopical findings of the kidney, the degenerative changes of endothelial cell, podocyte and mesangial cell in glomeruli were initially observed on the 10th water-deprived day as well as the degeneration of microvilli and intracellular organelle in the renal tubules.

• Journal of Life Science
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• v.29 no.9
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• pp.1034-1046
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• 2019

The mitochondria is the major cellular organelle of energy metabolism for the supply of cellular energy; it also plays an important role in controlling calcium regulation, reactive oxygen species (ROS) production, and apoptosis. Mitochondrial dysfunction causes various diseases, such as neurodegenerative diseases, Lou Gehrig's disease, cardiovascular disease, mental disorders, diabetes, and cancer. Most of the diseases are age-related diseases. In this review, we focus on the roles of mitochondrial dysfunction in cancer. Mitochondrial dysfunction induces carcinogenesis and is found in many cancers. The factors that cause mitochondrial dysfunction differ depending on the types of carcinoma, and those factors could cause cancer malignancy, such as resistance to therapy and metastasis. Mitochondrial dysfunction is caused by a lack of mitochondria, an inability to provide key substances, or a dysfunction in the ATP synthesis machinery. The main factor associated with cancer malignancy is mtDNA depletion. Mitochondrial dysfunction would leads to malignancy through changes in molecular activity or expression, but it is not known in detail which changes lead to cancer malignancy. In order to explore the relationship between mitochondrial dysfunction and cancer malignancy in detail, mitochondria dysfunctional cell lines are constructed using chemical methods such as EtBr treatment or gene editing methods, including shRNA and CRISPR/Cas9. Those mitochondria dysfunctional cell lines are used in the study of various diseases caused by mitochondrial dysfunction, including cancer.

• Applied Microscopy
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• v.15 no.2
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• pp.31-68
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• 1985

Authors are trying to unveil the ultrastructural organization of olfactory bulb, which has been summerized under light microscopic level or communicated only in some detail in different view point until now. For the critical point of view, since the phylogenetical approach will give the ultimate value in the correlative study between structural and functional bases (Brodal, 1969), the present study was carried out light and electron microscopic analyses of the structures of the neurons and synaptic organizations in olfactory bulbs from different animals in phylogenetical scale. We selected each one species from five animal classes: the house rabbit(Oryctolagus cuniculus var. domesticus [Gmelin]) from Mammalia, the domestic fowl (Gallus gallus domesticus Brisson) from Aves, the viper (Agkistrodon hylys [G.P. Pallas]) from Reptilia, a frog (Bombiana orientalis Boulenger) from Amphibia and the crussian carp (Carassius carassius [Linne]) from Pisces. For light microscopic study, samples were fixed in 10% formalin and paraffin sections were stained with hematoxylin-eosin. For the electron microscopic study, the tissues were fixed by perfusion through the heart or immersion with 1% paraform-aldehyde-glutaraldehyde mixture (phosphate buffer, pH 7.4), and final tissue block trimmed under dissecting microscope were osmicated (1% OsO4), they were embedded in Araldite or Epon 812, and ultrathin sections were made by LKB-V ultratome following the inspection of semi-thin sections stained with toluidine blue-borax solution. Ultra-thin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100CX electron microscope. We have summerized our morphological analyses as follows: 1. The olfactory bulb of rabbit, viper and frog shows the eight layers of fila olfactoria, glomerular, external granular, external plexiform, mitral cell, internal plexiform, internal granular, medullary but domestic fowl shows the five layers of glomerular, fibrillar, mitral, granular and medullary and the three layers of fibrilla, glomerular and medullary in crussian carp. The sharpness of demarcation between the layers shows deferential tendency according to phylogenetical order. 2. Mitral cells of vertebrate have large triangular or oval shape with spherical nuclei which contain not so much chromatin. The cytoplasm contains numerous cell organelles, of which Nissl's bodies or granular endoplasmic reticula arranged as parallel strands. Development of granular endoplasmic reticula were declined as the phylogentical grade is going lower. 3. Tufted cells of all animal are mostly spindle or polygonal contour and contain oval nuclei which located in periphery of cytoplasm. The nuclei of rabbit, fowl, viper and frog has relatively space chromatin, but a nucleus of crussian carp contain irregularly aggregated chromatin in karyoplasm. Their cytoplasmic volume and cell organelle contents are in between those of mitral cell and granular cell. They contain moderate amount of mitochondria, granular endoplasmic reticula, a few Golgi complex, polysomes, lysosome, etc. 4. Granule of cells of all the vertebrate amimals studied exhibit similar features; cells and their dense nuclei show spherical or oval contour, and they have the thin rim of cytoplasm which contain only a few cell organelles. 5. In rabbit, the soma of mitral cells were in contact with boutons with two types of synaptic vesicles, that is, round and flat vesicles, especially flat vesicles in boutons were showing reciprocal synapses. However, in domestic fowls, vipers, frogs and crussian carps, there were found boutons showing only spherical synaptic vesicles. 6. The boutons containing round synaptic vesicles were made contact with the some of tufted cell of olfactory bulb in the rabbits, fowls, vipers and frogs, but no synaptic boutons were observed in soma of tufted cells in crussian carps. In the frogs, there were observed dendrites were contact with the soma of tufted cells. 7. In the neuropils of plexiform, granular and glomerular layers olfactory bulbs in the vertebrate, the synapses were axo-large dendrites, axo-median and small dendrites, dendrodendritic, and axo-axonal contacts. However, in the neuropil of crussian carps, synapses were observed only in glomerular layer.