• The Korean Journal of Zoology
• /
• v.19 no.4
• /
• pp.143-154
• /
• 1976

The hemocytes of Drosophila melanogaster were observed with electron microscope, and five types of the cells were identified; prohemocyte, plasmatocyte, granular cell, crystal cell and oenocytoid, accounting for about 5%, 35%, 45%, 10%, 5% respectively of total cell numbers. Prohemocytes are characterized by a low concentration of intracellular organelles. Plasmatocytes are spindle or oval in shape and have relatively plenty of organelles and lysosomes. Granullar cells are the most polymorphic. They have numerous pseudopod-like projections and contain various granules and inclusions. In this cell type, intracellular organelles are fully developed. Crystal cells are characterized by numerous crystals composed of fine granules arranged regularly. Oenocytoids are the largest one among all cell types and contain relatively developed organelles.

• The Korean Journal of Oral and Maxillofacial Pathology
• /
• v.41 no.1
• /
• pp.1-7
• /
• 2017

Lycorine, a natural alkaloid extracted from the Amaryllidaceae plant family, was reported to various physiological and pharmacological effects including anti-cancer activity. Nevertheless, there is no report of the anticancer effect of lycorine in oral cancer cells. The effects of lycorine on cell proliferation and apoptosis were examined through trypan blue exclusion assay, 4'-6-diamidino-2-phenylindole (DAPI) stain, Live/Dead assay, Western blot analysis and RT-PCR. Lycorine suppressed cell viability and induced apoptosis in MC3 and HSC-3 cell lines. Lycorine decreased survivin protein but did not affect its mRNA. It regulated survivin through accelerating protein degradation in a time-dependent manner although neither proteasome nor lysosome was not associated with lycorine-mediated protein degradation. Collectively, our results suggest that lycorine may be a potential therapeutic anti-cancer drug candidate for the treatment of human oral cancer.

• Animal cells and systems
• /
• v.3 no.2
• /
• pp.221-228
• /
• 1999

In order to find a biochemical marker for late Iysosomes, we characterized two cDNAs which were cloned by using a monoclonal antibody (mAb) against Iysosomes in Amoeba proteus as a probe. The two cDNAs, a 1.3-kb cDNA in pBSK-Iys45 and a 1.6-kb cDNA in pBSK-Iys60, were found to encode proteins homologous to pepstatin-insensitive carboxyl proteinases (PICPs). E. coli transformed with pBSK-Iys45 produced two immunopositive polypeptides (45 and 43 kDa) and the cDNA in 1274 bases encoded a 44,733-Da protein (Lys45) of 420 amino acids containing one site for a core oligosaccharide. On the other hand, E. coli transformed with pBSK-Iys60 produced several polypeptides (64, 54, 45, 41, and 37 kDa) reacting with the mAb. The cDNA contained 1629 bases and encoded a 59,231-Da protein (Lys60) of 530 amino acids containing two sites for asparagine-linked core oligosaccharides. These two cDNAs showed identities of 60.3% in nucleotide sequences and 23.6% in amino acid sequences. Lys45 and Lys60 appeared to share XXEFQK as a common antigenic domain. The amino acid sequence of the Lys45 protein showed 17.4% identity and 40.9% similarity to that of PICP from Pseudomonas sp. 101. On the other hand, Lys60 showed a 24.3% identity and 51.9% similarity with human Iysosomal PICP in the amino acid sequence. A putative active center for serine protease, GTS＊xxxxxFxG, was found to be conserved among PICP homologues. The two PICPs are the first reported enzymatic markers for late Iysosomes.

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

• The Journal of the Korea Contents Association
• /
• v.9 no.12
• /
• pp.376-384
• /
• 2009

This research has microstructure observation to find tissue damage mechanism and radio-protection effect on mouse kidney tissue. The result observation of a Light Microscope(LM); The kidney tissue after 5Gy irradiation observed a glomerulus atrophy, also crack distance to base membrane of a convoluted tubules. The kidney tissue after 10Gy irradiation observed out flow cytoplasm to membrane break of a convoluted tubules. The result observation of a Transmission Electron Microscope(TEM); The kidney tissue of after 5Gy irradiation has to breaking a inside cristae and membrane of mitochondria, also show definite damage of nucleus membrane. 10Gy irradiation has all the more damage a base membrane and thickness of lysosome. However, Propolis eating groups observed normal to nucleus membrane and small body of intracellular. therefore We considered "Propolis" as make radio protection function to kidney tissue of the greater part.

• BMB Reports
• /
• v.29 no.3
• /
• pp.253-260
• /
• 1996

An enzyme that hydrolyzes flavin-adenine dinucleotide (FAD) was found to be present in rat liver lysosomal membrane prepared from Triton WR-1339 filled lysosomes (tritosomes) purified by flotation on sucrose. This FAD phosphohydrolase (FADase) exhibited optimal activity at pH 8.5 and had an apparent Km of approximately 3.3 mM. The activity was decreased 50~70% by dialysis against EDTA and this was restored by $Zn^{2+}$, $Mg^{+2}$, $Hg^{+2}$, and $Ca^{+2}$ ions inhibited the enzyme, but $F^-$ and molybdate had no effect. The enzyme was also inhibited by p-chloromercuribenzoate (pCMB), reduced glutathione and other thiols, cyanide, and ascorbate. The presence of ATP, ADP, AMP. ${\alpha}-{\beta}-methylene$ ATP, AMP-p-nitrophenyl phosphate (PNP), GMP, and coenzyme A (CoA) decreased the activity on FAD, but pyrimidine nucleotides, adenosine, adenine, or $NAD^+$ were without effect. Phosphate stimulated the activity slightly. FAD phosphohydrolase activity was separated from ATPase and inorganic pyrophosphatase activities by solubilization with detergents and polyacrylamide gel electrophoresis and by linear sucrose density gradient centrifugation suggesting that the enzyme is different from ATPase, inorganic pyrophosphatase, and soluble lysosomal FAD pyrophosphatase. Paper chromatography showed that FAD was hydrolyzed to flavin mononucleotide (FMN) and AMP which were further hydrolyzed to riboflavin and AMP by phosphatases known to be present in lysosomal membranes. Incubation of the intact Iysosomes with pronase showed that the active site of FAD phosphohydrolase must be oriented to the cytosol. The FAD hydrolyzing activity was detected in Golgi, microsome, and plasma membrane, but not in mitochondria or soluble lysosomal preparations.

• Applied Microscopy
• /
• v.28 no.3
• /
• pp.315-328
• /
• 1998

The present study was carried out to investigate the effect of ultrasonic bath in tissue preparation for transmission electron microscopy. The method used standard reagents and media, and employed ultrasonic bath agitation to accelerate fluid exchange. The liver kidney, stomach and cardiac muscle tissues of male Sprague-Dawley rats were used for the experiment, and the experimental design was divided into 4 groups; The control group using rotators (Traditional method, 1,625 mins) and the three experimental groups using ultrasonic bath (UB) in the primary fixation through the infiltration processes (UB I; 62.5 mins, UB II; 125 mins, UB III; 250 mins). The results were as follows; 1. In the control group, tissues were easily sectioned, and showed well preserved intact membranes, and cell organelles such as mitochondria, lysosome, peroxisome, rough endoplasmic reticulum and smooth endoplasmic reticulum. 2. In the UB treated group I, tissues showed holes due to the inadequate removal of both water and fluids used in the dehydration process. Also the mitochondria of cell organelles, especially, showed swollen intracristal spaces and dense matrices due to poor fixation. 3. In the UB treated group II, tissues showed good preservation of cell organelles and specimen slice sections. Also, no holes were observed. 4. In the UB treated group III, tissues showed leaching of structural components in the cytoplasm, but no holes were observed. In conclusion, the ultrasonic bath procedure takes approximately 120 minutes from specimen fixation to resin infiltration and gives excellent results.

• Journal of Life Science
• /
• v.28 no.6
• /
• pp.753-763
• /
• 2018

Cysteine cathepsins are lysosomal enzymes that belong to the papain family and can induce the degradation of damaged proteins through the endo-lysosomal pathway. It is highly upregulated in many cancers by regulating gene amplification and transcriptional, translational, and post-transcriptional modifications. Cathepsin S is part of the cysteine cathepsin family. Many studies have demonstrated that cathepsin S not only plays a specific role in MHC class II antigen presentation but also plays a crucial role in cancers. Cathepsin S is more stable at a neutral pH compared to other cysteine cathepsins, which supports the importance of cathepsin S in disease microenvironments. Therefore, the dysregulation of cathepsin S has participated in a variety of pathological processes, including cancer, arthritis, and cardiovascular disease. Furthermore, a decrease or depletion in the expression of cathepsin S has been implicated in the processes of tumor growth, invasion, metastasis, and angiogenesis. Taken together, cathepsin S has been suggested as an attractive therapeutic target for cancer therapy. In this review, the known involvement of cathepsin S in diseases, particularly with respect to recent work indicating its role in cancer therapy, is examined. An overview of current literature on the inhibitors of cathepsin S as a therapeutic target for cancer is also provided.

• Journal of Nutrition and Health
• /
• v.17 no.3
• /
• pp.224-235
• /
• 1984

Vitamin E status affected by dietary high PUFA and Se was examined by biochemical and morphological means. Rats were fed four different diets(I : 15% p/s=1 control diet, II : 15% perilla oil diet, III : 15% perilla oil, vitamin mix -vitamin E, IV : 15% perilla oil, vitamin mix-vitamin E and salt mix -Se ) for $4\frac{1}{2}$ weeks. Various dietary treatments had no significant effects on body weight gains of rats. Activities of serum creatine phosphokinase known as an indicator of vitamin E deficiency were significantly higher( P < 0.001) in groups fed diets high in PUFA, regardless of the addition or omission of vitamin E from the vitamin mixture than those in control group. Vitamin E concentrations of serum and liver were affected by experimental diets and serum levels were more affected than those in liver. Electron microscopic observations of the liver revealed 1) the presence of swollen and degenerated mitochondria and lysosome-like body(II), and 2) markedly swollen and degenerated mitochondria, numerous lysosomes and decreased in size and number of microvilli along the bile canaliculus ( III, and 3) a remarkable accumulation of lipid droplets, nuclear pyknosis, degenerated mitochondria and increased number of lysosomes scattered along the cell junction in the hepatocytes (IV).

• Clinical and Experimental Pediatrics
• /
• v.49 no.12
• /
• pp.1358-1362
• /
• 2006

Niemann-Pick disease is a group of autosomal recessive disorders associated with hepatosplenomegaly, variable neurologic deficits, and the storage of sphingomyelin and other lipids. Seven cases have been reported in Korea. We report an additional case presenting with hypotonia, early neurodevelopmental delay, hepatosplenomegaly and death by persistent pneumonia and asphyxia at the age of 23 months. MRI of brain and fundoscopic findings of our case at 4 months of age were normal. However, abnormal intensity of the thalamus and atrophy of the right temporal lobe on the MRI and macular cherry red spots were noticed at the age of 17 months. A bone marrow biopsy showed large foamy cells, while hexosaminidase A and B levels were normal. Although biochemical or molecular workup was not done, these findings led to the diagnosis of infantile onset Niemann-Pick disease, probably type A. A brief review of the related literatures was made.