• IMMUNE NETWORK
• /
• v.1 no.3
• /
• pp.221-229
• /
• 2001

Background: Inactivation of tumor suppressor genes is believed to be important in the development of many human malignancies. Recently, several lines of evidence have indicated that the wild type p53 gene located at 17p13.3, may function as a tumor suppressor gene and that a mutant p53 gene could promote transformation by inactivating normal p53 function in a dominant negative fashion. These broad spectrum of p53 mutation in human cancers provide that mutant p53 and their protein may be potential targets of tumor diagnostic and therapeutic interventions. Method: Colony formation was performed to investigate growth suppressional ability. p53 expression pattern was examined by western blot and p53-mediated transactivation ability was assessed by CAT activity. SNU C2A cells were observed in apoptotic aspects induced by etoposide and $H_2O_2$ treatment, detecting sensitivity on agent, DNA fragmentation through agarose gel, chromatin condensation by fluorescence microscope, and cell cycle distribution by FACS. Result: 1) p53 mutant his179arg ($histidine{\rightarrow}arginine$) detected in SNU C2A cells lost transcriptional activity and growth suppression ability, showing dominant negative effect on its wild type p53. 2) Etoposide-treated SNU C2A cells induced apoptosis, exhibiting dramatic reduction of cell growth, DNA fragmentation, nuclear condensation formation of apoptotic body and increment of sub-G1 cell fraction. 3) Etoposide and $H_2O_2$-treated SNU C2A cells have no high increase of p53 expression and overexpressed p53 protein changed localization, from cytoplasm to nucleus. Also, p53-mediated transcriptional activity was increased by agents-treatment. Conclusion: SNU C2A cells coexpress wild-type and mutant p53 protein induced apoptosis in the condition on DNA damage, through localizational shift from cytoplasm to nucleus of p53 protein rather than the induction of p53 protein. SNU C2A cells derived mutant p53 his179arg abrogated both the growth supression ability and transactivational activity, showing inhibition effect on transcriptional activity of wild type p53, but did not repress the activity of wild type p53 in SNU C2A cells owing to dominant activity of wild type. These cell condition may provide new gene therapeutic implications leading effective antiproliferation of cell when mutant and wild-type p53 protein were co-expressed in cell.

• Journal of Korean Society of Disaster and Security
• /
• v.11 no.2
• /
• pp.69-74
• /
• 2018

Fog to which environmental impacts are sensitive has a danger to the safety of citizens due to the difficulty in predicting the specific area/time zone. Therefore, we propose a white smoke fog reduction technique using a fog removal device that can remove fog particles directly through dry air and anionic condensation nucleus instead of conventional passive countermeasures. In this study, to verify the effect of reducing fog and the effect of temperature on the white smoke fog which is frequently occurred in the detention basin. As a result, the visible distance of 100m or more was secured within 30 seconds, and it was confirmed that the fog reduction effect is more effective. Also, the lower the temperature, the larger the amount of white smoke fog was. However, the effect of reducing the white smoke fog by temperature was insignificant. Through this experiment, it was possible to verify the reduction effect of the white smoke fog generated in the detention basin through fog removal device.

• Proceedings of the KSAR Conference
• /
• 2003.06a
• /
• pp.96-96
• /
• 2003

Nek2 (NIMA-related protein) is a mammalian cell cycle-regulated kinase that involves in chromosome condensation and centrosome regulation and NuMA (nuclear mitotic apparatus protein) is involved in spindle assembly during a cell cycle. The cellular distribution and organization of the centrosomal components is completely unknown during fertilization and embryonic development. We examined distribution of two well-known centrosomal proteins, Nek2 and NuMA in mouse gametes and embryos to get an insight in the reorganization of centrosomal proteins during germ cell development and early fertilization. Spermatogenic cells, gametes, and embryos were analyzed with anti-Nek2 or -NuMA antibodies by immunological assay, RT-PCR, and overexpression through gene transfection. Mitotically or meiotically active spermatogenic cells were intensively stained with these antibodies in both centrosomes and cytoplasm, whereas the oocytes showed different staining patterns depending on the meiotic stages. During maturation, GV, GVBD, and MI stage were clearly stained with NuMA antibody in the nucleus or cytoplasm at MII. Also, Nek2 was detectable in cytoplasm as scattered spots or chromosome associated at MII. In early developmental embryo, NuMA was detected in nucleus of each blastomere, while Nek2 was detected in cytoplasm. In contrast to previously reported results, Nek2 and NuMA were detected in both decondensing head, and the centriole of demembranated and decondensed sperm or whole body of trypsin-treated sperm for Nek2. During meiotic progress in oocytes, transcripts levels were the highest in MI stage and then downregulated in MII. Also, it shows dramatically change in early developmental embryos, firstly, it was increased until 4 cell stage and reduced in 8 cell stage, and finally, transcript levels were upregulated until blastoscyst. This finding suggests that cnetrosomal component may play an important role in reorganizing of functional centrosome during fertilization process and subsequent development.

• Restorative Dentistry and Endodontics
• /
• v.20 no.2
• /
• pp.549-576
• /
• 1995

Bacteria are one of the most important causative agents of the pulpal and periapical diseases. Streptococci are one of the most frequently isolated facultative anarerobic bacteria in the infected root canals. Bacterial cell wall components have a direct effect in the pathogenesis of the pulpal and periapical infections. Hyaluronidase produced by bacteria has been implicated in dissemination of the diseases. The purpose of this study was to evaluate the effect of cell wall extract of streptococci on the L929 cells using inverted microscope and the transmission electron microscopy (TEM). Hyaluronidase production of streptococcal strains were investigated to determine the correlation between the severity of cell damage and the activity of enzymes. Bacterial cell wall extracts of S. sanguis, S. mitis and S. uberis isolated from infected root canals and ATCC type strains of S. mutans (ATCC 10449) and E. faecalis (ATCC 19433) were prepared by sonication and confirmed with SDS-PAGE. Silver stain of SDS-PAGE of sonic extract was efficient at $100{\mu}g$/ml concentration of cell wall protein, while Coomasie blue stain was efficient at $100{\mu}g$/ml concentration. Inverted microscope showed that sonic extract-treated L929 cells were round and detached from the substratum while others lost their fibroblastic shapes. Transmission electron microscopic examination revealed that streptococcal extracts induced death of L929 cells. Sonic extracts of streptococci had variable effect on microstructure of L929 cells. significant chromatin condensation was observed in the nucleus of the cells. Disappearance of cell surface microvilli and nuclear fragments with dense chromatin were observed. The cell nucleus had an irregular shape and numerous large vacuoles were seen in the cytoplasm and some breaks of the cell membrane could be seen. Cell organelles were in various stages of destruction and cristae of mitochondria were disoriented or disappeared. Eighteen strains of streptococci did not produce hyaluronidase.

• Journal of Marine Life Science
• /
• v.6 no.1
• /
• pp.23-30
• /
• 2021

The differentiation process of male germ cells and sperm morphology of the abalone Haliotis discus hannai were described in ultrastructure. The differentiation process of sperm was divided into four stages: spermatogonium, spermatocyte, spermatid and sperm. The process of differentiation from spermatogonium to spermatocyte did not show significant morphological changes. However, during the spermiogenesis there were distinct morphological changes such as chromatin condensation, morphological changes of the nucleus, and formation of acrosome, midpiece and flagellum. The sperm of the abalone consisted of head, midpiece and tail. The head of approximately 5.3 ㎛ in length was composed of a nucleus of high electron dense and bullet-shaped acrosome. The midpiece was composed of the basal body and mitochondria, and five mitochondria were arranged in single layer around the basal body. The cross section of the tail showed a "9+2" axonemal structure. These morphological and structural features are the result of showing that the sperm of H. discus hannai is a primitive type.

• Journal of the Korean Chemical Society
• /
• v.63 no.6
• /
• pp.427-434
• /
• 2019

The role of hydrophobic residues on protein folding reaction was studied by folding kinetics measurements in conjunction with protein engineering. The HubWA, which was derived from human ubiquitin by mutating the residues at 45 (Phe to Trp) and 26 (Val to Ala), was used as a mutational background. Fourteen hydrophobic residues were mutated to alanine. Among fourteen variants generated, only four variant proteins (V5A, I13A, V17A, and I36A) were suitable for folding study. The folding kinetics of these variants was measured by stopped-flow fluorescence spectroscopy. The folding kinetics of HubWA and V17A was observed to follow a three-state on-pathway mechanism. On the other hand, folding kinetics of V5A, I13A, and I36A was observed to follow a two-state mechanism. Based on these observations, transition of protein folding reaction from collision-diffusion mechanism to nucleation-condensation mechanism was discussed.

• Applied Microscopy
• /
• v.37 no.3
• /
• pp.185-198
• /
• 2007

Spermiogenesis in Japanese white-toothed shrew. Crocidura dsinezumi was investigated by transmission electron microscope. Spermiogenesis was divided into 12 phases 14 steps, based on the morphological features of the nucleus and change of organelles in cytoplasm. The nucleus of spermatids in Golgi (step $1{\sim}2$) phase were spherical; however, they were changed into oval in the cap (step $3{\sim}6$) phase. Flagellum appeared in the middle of acrosomal phase; on the other hand, slender and long spermatid head was formed in maturation phase. The head of spermatids faced the lumen in step 1 to step 6 (from Golgi to cap phase), but, in step 7 to step 14 (from acrosomal to spermiation phase), it turned its head to the basal lamina of the seminiferous epithelium. The nucleus and acrosome were elongated maximally in step 10. The condensation of chromatin started in late acrosomal (step 10) phase, and it was completely finished and homogenized in the middle of maturation (step 12) phase. Multivesicular body appeared near the acrosomal vacuole during the middle cap (step 5) phase, and a large number of them were observed near the Golgi apparatus in the late cap (step 6) phase. Considering all the results, the spermiogenesis might be useful information to analyse the differentiation of spermatogenic fells.

• Applied Microscopy
• /
• v.12 no.1
• /
• pp.41-47
• /
• 1982

Mycosis fungoides is an uncommon, chronic fatal disease of lymphoreticular system associated with primary ski3 involvement for many years and terminating as a malignant lymphoma with involvement of lymph nodes and viscerae. On occasion it simulates numerous other nonspecific benign skin lesions, thus it may be impossible to decide whether the infiltrate represents early mycosis fungoides or nonspecific on the histopathologic ground alone. A case of mycosis fungoides was confirmed by electron microscopy and reported here. The patient was 69-years-old male who had suffered from erythematous scaly eruption on the whole body since 10 years. Skin biopsies of 4 times showed focal ulceration with chronic nonspecific inflammation and polymorphic cell infiltration in lower dermis, thus possibility of mycosis fungoides could not be completely ruled out. Electron microscopically several atypical lymphoid cells, which had a large cerebriform nucleus with peripheral condensation of dense chromatin and scant cytoplasm, were noted in the upper dermis. Intraepidermal infiltration of these atypical cells was also seen. It was thought that the electron microscopic study may be very helpful to differentiate equivocal mycosis fungoides from the nonspecific dermatosis.

• Journal of the Korean Chemical Society
• /
• v.56 no.2
• /
• pp.246-250
• /
• 2012

An efficient access, single step and environmentally benign synthesis of a new series of pyrazole containing isoxazolines derivatives were prepared by the condensation of chalcones bearing pyrazole moiety with hydroxyl amine hydrochloride in basic condition by using polyethylene glycol-400 (PEG) as a greener reaction solvent. The advantages of the present methodology are mild reaction condition and avoidance of volatile organic solvent. Furthermore, these newly synthesized compounds were screened for their antimicrobial activity against various pathogens like Escherichia coli (MTCC 2939), Salmonella typhi (MTCC 98), Staphylococcus aureus (MTCC 96), Bacillus subtilis (MTCC 441), Aspergillus niger (MTCC 281), Aspergillus flavus (MTCC 2501), Penicillium chrsogenum (MTCC 160) and Fusarium moniliformae (MTCC 156). Especially compound containing the hydroxyl group in C2-position and presence of halo (I, Br and Cl) groups as substituents at $C_3$ and $C_5$ position on the benzene nucleus showed the higher activity. Furthermore, compounds bearing methyl groups in combination with I and Br which enhanced the activity.

• Restorative Dentistry and Endodontics
• /
• v.21 no.1
• /
• pp.385-402
• /
• 1996

Cytotoxicity of dental restorative materials using cell culture technique has been extensively studied by various quantitative assays. The aim of this study was to investigate the microstructural change of damaged L292 cells which could not observed with light microscope. Cytotoxic effect of ZOE, Prisma APH (Densply International Inc., U.S.A.), Clearfil FII(Kuraray Co., Japan), Fuji II(GC Co., Japan) and Fuji II LC(GC Co., Japan) on cultured L292 cells were observed. Irreversible cell damage and cytolysis were found in ZOE and Fuji II groups. In Clearfil FII, mild to moderate cell damage was observed. APH group showed variable cytotoxicity. Moderate cell damage was found in Fuji II LC group. Cytotoxic effect were as follows : A condensation of the chromatin occureds along or adjacent to the inner membrane of the nuclear envelops. The nuclear envelope remained resonably intact but the contents were partially or completely lost. The cell nucleus contains clusters of markedly electron-dense interchromatin granules. The rough endoplasmic reticulum were dilated. In some mitochondira, matrix was disoriented and fused cristae were discernible. Mitochondiral swelling and woolly appearance were recognized. Large vacuoles and autolysosmes were found in cytoplasm. Some breaks of the cytoplasmic membrane and even cytolysis could be seen in dying cells.