• Tuberculosis and Respiratory Diseases
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• v.42 no.5
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• pp.646-653
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• 1995

Background: The confirmative diagnosis of pulmonary tuberculosis(Tb) can be made by the isolation of Mycobacterium Tuberculosis(MTb) in the culture of the sputum, respiratory secretions or tissues of the patients, but positive result could not always be obtained in pulmonary Tb cases. Although there are many indirect ways of the diagnosis of Tb, clinicians still experience the difficulty in the diagnosis of Tb because each method has its own limitation. Therefore development of a new diagnostic tool is clinically urgent. It was reported that silica cause some lysosomal enzymes to be released from macrophages in vitro and one of these enzymes is elevated in workers exposed to silica dust and in silicotic subjects. In pulmonary Tb, alveolar macrophages are known to be activated after ingestion of MTb. Activated macrophages can kill MTb through oxygen free radical species and digestive enzymes of lysosome. But if macrophages allow the bacilli to grow intracellularly, the macrophages will die finally and local lesion will enlarge. Then it is assumed that the lysosomal enzymes would be released from the dead macrophages. The goal of this investigation was to determine if there are differences in the plasma activities of lysosomal enzymes, ($\beta$-glucuronidase(GLU) and $\beta$-N-acetyl glucosaminidase(NAG), among the groups of active and inactive pulmonary Tb and healthy control, and to see if there is any possibility that the plasma activity of GLU and NAG can be used as diagnostic indicies of active pulmonary Tb. Methods: The plasma were obtained from 20 patients with bacteriologically proven active pulmonary Tb, 15 persons with inactive Tb and 20 normal controls. In 10 patients with active pulmonary Tb, serial samples after 2 months of anti-Tb medications were obtained. Plasma GLU and NAG activities were measured by the fluorometric methods using 4-methylumbelliferyl substrates. All data are expressed as the mean $\pm$ the standard error of the mean. Results: The activites of GLU and NAG in plasma of the patients with active Tb were $21.52{\pm}3.01$ and $325.4{\pm}23.37$(nmol product/h/ml of plasma), respectively. Those of inactive pulmonary Tb were $24.87{\pm}3.78$, $362.36{\pm}33.92$ and those of healthy control were $25.45{\pm}4.05$, $324.44{\pm}28.66$(nmol product/h/ml of plasma), respectively. There were no significant differences in the plasma activities of both enzymes among 3 groups. The plasma activities of GLU at 2 months after anti-Tb medications were increased($42.18{\pm}5.94$ nmol product/h/ml of plasma) in the patients with active pulmonary Tb compared with that at the diagnosis of Tb(P-value <0.05). Conclusion: The results of the present investigation suggest that the measurement of the plasma activities of GLU and NAG in the patients with active pulmonary Tb could not be a useful method for the diagnosis of active Tb. Further investigation is necessary to define the reasons why the plasma activities of the GLU was increased in the patients with active pulmonary Tb after Tb therapy.

• Journal of Aquaculture
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• v.6 no.1
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• pp.1-12
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• 1993

Identification of blood cells and their physological functions in the cultured scallop, Patinopecten yessoensis collected from Abashiri Bay, Hokkaido, Japan were studied by electron microscopic structure and histological observations. The physiological function of each blood cell type was studied on the basis of its cytological structure, the lysosome in the blood cell and its phagocytosis. The blood cells were classified as Type I, Type II and Type III. The morphological characteristics of each blood cell type are as follows ; Type I : The cell is oval shaped and its cytoplasm contains comparatively low electron dense materials. The oval nucleus is sometimes ramified into two nuclei. Lumps of tubular smooth endoplasmic reticula and vacuoles are distributed near the nucleus. Type II : The cell appears long and oval shaped, and its cytoplasm contains high electron dense materials. The oval nucleus does not ramify, and large numbers of sac-like smooth endoplasmic reticula and free ribosomes are developed around the nucleus. No vacuoles exist in the cytoplasm Type 1II : The cell is round in shape and the electron density of the cytoplasm is the highest among the three types of cells because of large quantities of rough­surfaced endoplasmic reticula and no vacuoles. Particularly, the nucleus reveals a wheel-like shape owing to lumps of tuberous chromatin. The cells of Type I and II seem to have the role of carrying out phagocytosis on either foreign materials such as bacteria or endogenous old cells and the transport of nutritive materials. The type III cell, which has not been found in any bivalve species of non Pectinidae, may be said to have the function of production and the secretion of protein related to some humoral defense materials.

• Applied Microscopy
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• v.39 no.2
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• pp.107-115
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• 2009

The programed cell death of the cutaneous epithelial tissue during tail regression stages in anuran tadpoles of the blackspotted frog, Rana nigromaculata were visualized by the histochemical and transmission electron microscopic techniques. Metamorphotic changes in the tail regression during the period of the Shumway stage number 31 to 33 are characterized by the disappearance of mucous layer and formation of compound epithelium through cutaneous thickening. Following the TUNEL (terminal deoxynucleotidyl transferase-mediated biotinylated d-uridine triphosphate nick end labeling) staining technique, the apoptotic cells were detected at the distal region of the tail skin initially, but they can be seen at the proximal region according to their following development. It has been also revealed that the number of the TUNEL-positive cells gradually increased from apical to basal direction of the epithelial layers during the tail regressing stages. Following the TEM observation, the early apoptotic cells shown in the epithelium demonstrated condensation and margination of the chromatin material at the nuclear periphery. Another epithelial apoptotic cells were shown nuclear fragmentation, membrane blebbing and cytoplasmic condensation. Following the process of the apoptotic degradation, well preserved organelles and nuclear fragments can be identified in the cytoplasm of lysosome-rich cells, however they soon reduced to lysosomal residual bodies through the progressive degradation.

• Parasites, Hosts and Diseases
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• v.56 no.5
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• pp.409-418
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• 2018

Acanthamoeba spp. are free-living protozoa that are opportunistic pathogens for humans. Cysteine proteases of Acanthamoeba have been partially characterized, but their biochemical and functional properties are not clearly understood yet. In this study, we isolated a gene encoding cysteine protease of A. castellanii (AcCP) and its biochemical and functional properties were analyzed. Sequence analysis of AcCP suggests that this enzyme is a typical cathepsin L family cysteine protease, which shares similar structural characteristics with other cathepsin L-like enzymes. The recombinant AcCP showed enzymatic activity in acidic conditions with an optimum at pH 4.0. The recombinant enzyme effectively hydrolyzed human proteins including hemoglobin, albumin, immunoglobuins A and G, and fibronectin at acidic pH. AcCP mainly localized in lysosomal compartment and its expression was observed in both trophozoites and cysts. AcCP was also identified in cultured medium of A. castellanii. Considering to lysosomal localization, secretion or release by trophozoites and continuous expression in trophozoites and cysts, the enzyme could be a multifunctional enzyme that plays important biological functions for nutrition, development and pathogenicity of A. castellanii. These results also imply that AcCP can be a promising target for development of chemotherapeutic drug for Acanthamoeba infections.

• The Korean Journal of Zoology
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• v.28 no.4
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• pp.211-226
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• 1985

The ultrastructure of the digestive organ of Korean Planaria (Dugesia japonica) is studied by light microscope and transmission electron microscope. 1. Pharynx The epithelium surrounding pharyngeal lumen has a number of microvilli on the free surface. The epithelial cells contain PAS-positive granules which are 0.4 to 0.6 $\\mum$ in size. They also contain hundreds of vesicles and vacuoles. The pharyngeal epithelium of the external surface surrounded by pharyngeal cavity possesses a number of cilia and microvilli on the free surface. A number of muscle bundles are found in the pharyngeal tissue. The parietal epithelium surrounding pharyngeal cavity have microvilli and electron-dense secretory granules. 2. Caeca The cells which constitute the cecal epithelium are divided into four kinds of cells. 1) Phagocytic cell : These cells are characterized by presence of a number of lysosomes. These cells have highly developed mitochondria, polyribosomes and granular endoplasmic reticulum of which cisternae are distended. 2) Granular club cell : These cells contain round granules 5 $\\mum$ in diameter which show strong PAS-positivity and weak eosinophilia. The cells have highly developed granular endoplasmic reticulum. 3) Storage cell : These cells include thousands of glycogen granules in the cytoplasm. These cells also have second kind of round granules which are 1.4 to 3 $\\mum$ in size and exhibit PAS-positive reaction. 4) Immature storage cell : These cells have a large nucleus and contain a small number of granules which have PAS-positive granules and a few lipid droplets. Several chromatoid bodies are found in the cytoplasm around the nucleus.

• The Korean Journal of Zoology
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• v.29 no.3
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• pp.215-233
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• 1986

The effect of tryptophan on brain formation at the early stage of chick embryo has been investigated morphologically using electron microscope. The electron micrographs of cerebral cortex cells of $5\\sim10$ day old chick embryo, which received 1.0mg of tryptophan showed that the irregularity, evagination and disruption of nuclear membrane and nuclear chromatin condensation, nucleolar chromatin margination and segragation. Hypertrophy of stalks, vesicles, and vacuoles can be seen and dilation and vesiculation of rough endoplamic reticulum and polysome disaggregation occured. Protein and RNA levels and the activity of several enzymes such as lactate dehydrogenase, succinate dehydrogenase, malate dehydrogenase and glucose 6-phosphate dehydrogenase of tryptophan administered group were significantly lower than those of control group suggesting that the tryptophan administration depressed protein biosynthesis resulting in the decrease of enzyme activity. It was found that serotonin content of egg yolk which has been incubated for 10 days were as much as three times that of control egg yolk. It is not clear whether the increase of serotonin content might inhibit intracellular yolk granule degradation which might result in malformation of chick embryo, but it is likely that tryptophan administration might depress protein biosynthesis, consequently, the enzyme biosynthesis would be impaired. This might give rise to improper development of chick embryo.

• The Journal of the Korea Contents Association
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• v.10 no.7
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• pp.297-304
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• 2010

As Small Intestine Absorptive Epithelium Cells are surrounded by mucus polysaccharide and lymphocytes and mitochondria, they are sensitive to radiation energy. Damaged cells lead to a deficiency of nutrients and the imbalance of electrolyte metabolism, which in turn can becomes a major cause of an intestine tract death. This research observed ultra structures after injecting propolis into the abdominal cavity in order to reveal the radiation damage mechanism and radioprotection effect of intestine absorptive epithelium cells. The result of this research's observation found that stenosis occurred in the small intestine in some tissues 20 days after 5Gy irradiation, their surface turned black, and their elasticity dropped. Through observation with an optical microscope, it was found that the size of the goblet cells decreased, while the paneth granulate cells atrophied and were vacuolated. Observation with an transmission electron microscope(TEM) revealed that while microvill and lysosome were normally observed in jejunum tissues, mitochondria membrane was damaged and uneven surfaces were formed on lymphocytes. The membrane of absorptive epithelium cells hypertrophied in tissues of the ileum, and vacuole was observed. However, the observation after injecting propolis into the abdominal cavity found that mitochondria damage dropped dramatically, and radioprotection effects were to some extent confirmed, considering that glycocalyx of villi was clear, and M cells could be observed.

• Applied Microscopy
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• v.3 no.1
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• pp.1-16
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• 1973

A combined cytochemical and electron microscopic study was carried out for the demonstration of acid phosphatase activities in trophozoites of E. histolytica. and E. gingivalis. E. histolytica(YS-27) strain was isolated from liver abscess of 72-year-old man in September 1969, and E. gingivalis (YS-215) strain was collected from gingival crevice of 41-year-old man in January 1972. The amoeba strains were maintained by subculture on diphasic medium, and used throughout the study. The results are summarized as follows; 1. In E. histolytica, the reaction products were distributed evenly over the entire surface of plasma membrane, whereas E. gingivalis showed no activity of acid phosphatase on the plasma membrane, except in the portion of the uroid-like structure. 2. In the cytoplasm, various reaction precipitates were observed in vacuoles of both amoebae; vacuole limiting membrane, vacuole membrane and its contents and lysosome-like structure. Strong enzyme active contents but membrane reaction negative vacuoles were conspicuous in E. gingivalis. Endoplasmic reticulum showed a moderate activity. 3. Granule-like acid phosphatase reaction product was demonstrated in the nucleoplasm of E. gingivalis, but it was negative in E. histolytica.

• Microbiology and Biotechnology Letters
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• v.29 no.2
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• pp.90-95
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• 2001

Kinetic Analysis of Cathepsin B Inhibitor Using a Spectrophotometric Assay. Han, Kil-Hwan and SangDal Kim*. Department of Applied MicrobioJ0f5Yt Yeungnam UniversitYt Kyongsan 77 2-749, Korea - The KHS 10, C4Hl10~6 formula produced from Streptomyces luteogriseus KT-] 0 effectively inhibited a lysosomal cysteine proteinase, cathepsin B. It inhibited the enzyme activity of cathepsin B competitively when the N a-CBZ-Llysine p-nitrophenyl ester HC] (CLN) was used as a substrate. The inhibition const:mt (Ki) of KHS 1 0 for cathepsin B detennined by spectrophotometeric assay was 430 nM. The effective inhibition of cathepsin B was observed at $25^{\circ}C$ :md pH 6.0. The cathepsin B inhibitor, KHSlO needed a preincubation of cathepsin B with the inhibitor for over 5 min. The KHS 10 preserved over 80% inhibition activity even after heat-treatment at $100^{\circ}C$ for ] hr.

• The Korean Journal of Physiology and Pharmacology
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• v.15 no.1
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• pp.1-7
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• 2011

Many anticancer agents as well as ionizing radiation have been shown to induce autophagy which is originally described as a protein recycling process and recently reported to play a crucial role in various disorders. In HCT116 human colon cancer cells, we found that curcumin, a polyphenolic phytochemical extracted from the plant Curcuma longa, markedly induced the conversion of microtubule-associated protein 1 light chain 3 (LC3)-I to LC3-II and degradation of sequestome-1 (SQSTM1) which is a marker of autophagosome degradation. Moreover, we found that curcumin caused GFP-LC3 formation puncta, a marker of autophagosome, and decrease of GFP-LC3 and SQSTM1 protein level in GFP-LC3 expressing HCT116 cells. It was further confirmed that treatment of cells with hydrogen peroxide induced increase of LC3 conversion and decrease of GFP-LC3 and SQSTM1 levels, but these changes by curcumin were almost completely blocked in the presence of antioxidant, N-acetylcystein (NAC), indicating that curcumin leads to reactive oxygen species (ROS) production, which results in autophagosome development and autolysosomal degradation. In parallel with NAC, SQSTM1 degradation was also diminished by bafilomycin A, a potent inhibitor of autophagosome-lysosome fusion, and cell viability assay was further confirmed that cucurmin-induced cell death was partially blocked by bafilomycin A as well as NAC. We also observed that NAC abolished curcumin-induced activation of extracelluar signal-regulated kinases (ERK) 112 and p38 mitogen-activated protein kinases (MAPK), but not Jun N-terminal kinase (JNK). However, the activation of ERK1/2 and p38 MAPK seemed to have no effect on the curcumin-induced autophagy, since both the conversion of LC3 protein and SQSTM1 degradation by curcumin was not changed in the presence of NAC. Taken together, our data suggest that curcumin induced ROS production, which resulted in autophagic activation and concomitant cell death in HCT116 human colon cancer cell. However, ROS-dependent activation of ERK1/2 and p38 MAPK, but not JNK, might not be involved in the curcumin-induced autophagy.