• Research in Plant Disease
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• v.17 no.1
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• pp.78-81
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• 2011

Soft rot caused by Rhizopus oryzae occurred on unshiu orange (Citrus unshiu Marc.) sampled from commercial markets in Jinju, Korea, 2010. The first symptom of soft rot on orange is a water-soaked appearance of the affected tissue. The infected parts later disintegrated into a mushy mass of disorganized cells followed by rapid softening of the diseased tissue. The lesion on orange was rapidly softened and rotted, then became brown or dark brown. Optimum temperature for mycelial growth of the causal fungus on potato dextrose agar was $30^{\circ}C$ and growth was still apparent at $37^{\circ}C$. Sporangiophores were $6{\sim}20\;{\mu}m$ in diameter. Sporangia were globose and $40{\sim}200\;{\mu}m$ in size. The color of sporangia was brownish-grey to blackish-grey at maturity. Sporangiospores were sub-globose, brownish- black streaked and $4{\sim}10\;{\mu}m$ in size. Columella were globose to sub-globose and $85{\sim}120\;{\mu}m$ in size. On the basis of mycological characteristics, pathogenicity test, and the ITS sequence analysis, the causal fungus was identified as Rhizopus oryzae. To our knowledge, this is the first report of soft rot caused by R. oryzae on unshiu orange in Korea.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.26 no.3
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• pp.254-261
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• 2000

The present study has attempted to look into the mechanism of ras-induced carcinogenesis in a human epithelial cell system. Human epithelial cells immortalized with Ad12-SV40 hybrid virus were used to assess carcinogenic potential of the ras-oncogene. Cells transfected with pSV2-ras showed characteristics of cellular transformation. The transformation parameters such as cell density, soft-agar colony formation, and cell aggregation were significantly increased in the cells expressing ras oncoprotein. In addition, the duration required for the appearance of foci was shortened in the ras-transfected cells. Consistent with other reports, our results demonstrated an evidence that the ras-oncogene induced the cellular transformation of human epithelial cell system. When a high concentration of glucocorticoid was added into the media, transformation process was accelerated. It is speculated that glucocorticoid may provide an advantageous environment for the proliferation of the transformed cells. The induction of the intracellular free calcium concentrations following agonist treatment was significantly lower in the transformed cells than in the control cells. These effects were more manifested in the presence of extracellular cacium, indicating that the transformation process may alter the influx pathway of extracellular calcium. The induction of $IP_3$ following agonist treatment was also lower in the transformed cells than in the control cells. Thus, it is suggested that phospholipase C-coupled pathway was down-regulated in the process of the ras-induced transformation. While the levels of $TGF-{\beta}_1$ and PAI-2 mRNAs were decreased, the level of fibronectin mRNA was increased. The results indicate that mechanism of the ras-induced transformation may be associated with the altered expressions of growth regulatory factors. The present study demonstrates an evidence that the ras-induced cellular transformation may be associated with alteration of signal transduction and growth regulatory factors. The study will contribute to improve the understanding of molecular mechanism of epithelium-derived cancers including oral cancer.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.3
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• pp.218-228
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• 2010

Components of dental resin-based restorative materials are reported to leach from the filling materials even after polymerization. Hydroquinone (HQ) is one of the major monomers used in the dental resin and is known as a carcinogen. Thus, carcinogenic risk of HQ leaching from the dental resin becomes a public health concern. The present study attempted to examine the carcinogenic potentials of HQ on the human epithelial cell, which is the target cell origin of the most of oral cancers. Cytotoxicity of HQ was observed above 50${\mu}M$ as measured by LDH assay, indicating a relatively low toxicity of this substance in human epithelial cells. The parameters of neoplastic cellular transformation such as cell saturation density, soft agar colony formation and cell aggregation were analyzed to examine the carcinogenic potential of HQ. The study showed that 2-week exposure of HQ showed the tendency of increase in the saturation density and the significant enhancement of soft agar colony formation at the highest dose, 50 ${\mu}M$ only. It is suggested that HQ has a weak potential of carcinogenicity. When cells were treated with HQ and TPA, a well-known tumor promoter, the parameters of neoplastic cellular transformation was significantly increased. This result indicates that the potential risk of carcinogenicity from HQ is largely dependent upon the presence of promoter. Exposure of 50 ${\mu}M$ HQ increased the time-dependent apoptosis as measured by the ELISA kit. This concentration coincides with a dose of neoplastic transformation, indicating a possible link between apoptosis and HQ-induced cellular transformation. Hydroquinone generated Reactive Oxygen Species (ROS) which was evidenced by the treatment of antioxidants such as trolox and N-acetyl cysteine and the GSH depleting agent, BSO. Antioxidants blocked the generation of ROS and the GSH depleting agent, BSO dramatically increased the ROS production. Since HQ is known to increase ROS production thru activation of transcriptional factor such as c-Myb and Pim-1, it is speculated that ROS generation by HQ plays a role in the activation of oncogene, which may lead to neoplastic transformation. In addition, ROS is involved in the alteration of signal transduction, which regulates the apoptosis in many cellular systems. Thus, ROS-mediated apoptosis may be involved in the HQ-induced carcinogenic processes. Protein kinase C (PKC) is known to play pivotal roles in neoplastic transformation of cells and its high expression is often found in a variety of types of tumors including oral cancer. PKC translocation of PKC-${\alpha}$ was observed following HQ exposure. Altered signaling system may also play a role in the transformation process. Taken together, HQ leached from the dental resin does not pose a significant threat as a cancer causing agent, but its carcinogenic potential can be significantly elevated in the presence of promoter. The mechanism of HQ-induced carcinogenesis involved ROS generation, apoptosis and altered signaling pathway. The present study will provide a valuable data to estimate the potential risk of HQ as a carcinogen and understand mechanism of HQ-induced carcinogenesis in human epithelial cells.

• Journal of Dental Rehabilitation and Applied Science
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• v.33 no.2
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• pp.106-113
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• 2017

Purpose: The purpose of this study was to evaluate tissue dissolving capacity, antimicrobial effect of Hydroxyethylidene bisphosphonate (HEBP) interacting with sodium hypochlorite (NaOCl), Ethylenediaminetetraacetic acid (EDTA) as conventional endodontic irrigants and to determine tissue dissolving efficacy depended on temperature. Materials and Methods: A total of 80 bovine muscles were randomly distributed into 8 groups (n = 10). After their initial weights determined on a precision scale, the specimens in each group were immersed in the solutions for 5, 10 and 15 min and reweighted at each time period. Agar diffusion test inoculated with Enterococcus faecalis was performed for antimicrobial effect of each endodontic irrigants. Results: The ability to dissolve organic matter was greater in NaOCl group following NaOCl and HEBP mixture. Heated NaOCl ($40^{\circ}C$) and NaOCl/ HEBP mixture was greater tissue dissolving efficacy than room temperature ($25^{\circ}C$). Antimicrobial effect was greater and significant in the following order EDTA > EDTA + 1% NaOCl > $1%\;NaOCl{\geq}1%\;NaOCl$ + HEBP. Conclusion: HEBP as soft chelating agent does not disturb antimicrobial effect and less affected tissue dissolving efficacy as inherent properties of NaOCl. In the heated NaOCl/HEBP mixture analyzed, it dissolved more the organic matter than room temperature.

• Applied Biological Chemistry
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• v.42 no.1
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• pp.20-28
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• 1999

To improve Bacillus strains producing biopolymer, conditions for protoplast formation and regeneration were investigated in biopolymer producing Bacillus subtilis K-1 and lactose utilizing Bacillus coagulans. Bacillus subtilis K-1 mutant (SM-2) and Bacillus coagulans mutants (CM-12) were marked auxotrophic and antibiotics-resistant (SM-2) and an antibiotics-resistant mutants, respectively. To formate protoplasts derived from the mutants, conditions were established as follows. For B. subtilis mutant SM-2, its culture in mid-logarithmic phase was added with penicillin G (1.0 unit/ml) and further reacted for 1.5 hr. Cells were collected and then treated in lysis fluid (pH 7.0) containing 0.4 M sucrose and lysozyme $25\;{\mu}g/ml$ for 40 min at $37^{\circ}$. Protoplast formation was very successful (99.6%) and the ratio of cell wall regeneration was 2.4%. For Bacillus coagulans mutant CM-12, its mid-logarithmic phase culture was treated with penicillin G (0.3 unit/ml) and glycine (0.5%) for 1hr. Cells were collected and then resuspended in lysis buffer (pH 7.0) containing 0.6 M lactose and lysozyme $(300\;{\mu}g/ml)$ for 30 min at $37^{\circ}$. Protoplast formation was also successful (90.8%) and cell wall regeneration ratio was similar to SM-2 (2.2%). To improve regeneration frequency, regeneration medium was obtained as followed condition,. Cell wall regeneration was improved 2-4 folds with 5.1% for B. subtilis SM-2 and 10.3% for B. coagulans CM-12 when protoplasts mixed with soft top agar(0.4%) was overlaid onto trypticase soy broth medium containing 0.4 M sucrose, 0.7% casamino acid, 1% PVP, 25 mM $MgCl_2,\;25\;mM\;CaCl_₂$ and 1.5% agar.

• Research in Plant Disease
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• v.20 no.1
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• pp.50-53
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• 2014

Rhizopus soft rot of lily (Lilium longiflorum) caused by Rhizopus oryzae was observed in the experimental field in Taean Lily Experiment Station in Korea, 2012. The typical symptoms were water-soaked lesions on bottom stem and leaf rot. The lesion rapidly expanded and the plant was softened totally. The fungus grew vigorously at an optimum temperature ($25^{\circ}C$) and brownish colony and black sporangia were formed on potato dextrose agar medium. Sporangiophores formed on end of sporangia were sub-globose, brownish and $6-10{\mu}m$ in size. Sporangia were globose, blackish and $87-116{\mu}m$ in size. Sporangiospores were irregularly oval and sub-globose, brownish $4-8{\mu}m$ in size. On the basis of mycological characteristics, analyzing sequences of internal transcribed spacer region of ribosomal DNA, and pathogenicity test on host plants, the causal fungus was identified as R. oryzae. This is the first report of Rhizopus soft rot on lily caused by R. oryzae in Korea.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.7
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• pp.1055-1059
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• 2003

Keratins, the constituents of epithelial intermediate filaments, are precisely regulated in a tissue and development specific manner. There are two types of keratin in bovine. The type I is acidic keratin and the type II is neutral/basic keratin. 1.5 kb of 5' flanking sequence of Korean cattle Keratin IV gene, type II keratin (59 kDa), was cloned and sequenced. A symmetrical motif AApuCCAAA are located in a defined region upstream of the TATA box. Proximal SP1, AP1, E-box and CACC elements as the major determinants of transcription are identified. When it was compared to the bovine sequence from -600 bp to ATG upstream, the homology was 97% in nucleotide sequence. Several A and T sequences, located in the promoter region, are deleted in the Korean cattle. An expression vector consisted of Korean cattle Keratin IV gene promoter/SV40 large T antigen was transfected to HaCaT cell (Epithelial keratinocyte). The transformed HaCaT cells showed active proliferation when treated with PDGF (Platelet-derived growth factor) in 0.3% soft agar compared to control cells. These results indicate that Korean cattle Keratin IVgene promoter can be used as a promoter for transfection into epithelial cell.

• Journal of Veterinary Clinics
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• v.16 no.1
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• pp.26-30
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• 1999

본 연구는 임상형 및 준임상형 유방염에 이환된 젖소의 유즙에서 분리된 24주의 황색포도상구균에 대한 capsular polysaccharide(CP)형을 확인하고, 1% 토끼 혈청이 함유된 serum-soft agar 배지에서 집락의 모양을 관찰하였다. 또한 동물에 따라 우세한 CP형의 차이가 있는지를 비교하고자 개에서 분리된 13주에 대하여 동일한 실험을 수행하여 다음과 같은 결과를 얻었다. 집락모양을 관찰한 결과 젖소 유래 24주 중 16주(66.7%)는 diffuse, 5주(20.8%)는 compact, 나머지 3주(12.5%)는 분류가 불가능(indeterminate)하였다. CP형 확인결과 9주(37.5%)는 type 5, 2주(8.3%)는 type 8, 13주(54.2%)는 분류가 불가능(non-typeable)하였다. 한편, 개에서 분리된 균주 중 1주(type 8)를 제외한 12주(92.3%)는 type 5로 분류되었으며, 13주 중 8주(61.5%)가 diffuse형의 집락을 보였다. 본 실험에 사용한 균주의 수가 충분하지 못한 문제가 있지만 동물에 따라 우세한 CP형이 다를 수 있으며 분류 불가능한 균주가 상대적으로 높은 비율을 차지하였는데 이는 새로운 CP형의 분류형이 필요함을 시사하는 것으로 판단된다. 또한 type 5와 type 8만을 포함한 유방염 백신은 한계가 있을 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.21
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• pp.9131-9136
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• 2014

ELDF15, homologous with AT2 receptor-interaction protein 1 (ATIP1), may play an important role in cell differentiation, proliferation, and carcinogenesis. We aimed to understand the biological function of ELDF15 via construction and transfection of a recombinant expression vector containing antisense ELDF15. Recombinant expression vectors were successfully constructed and transfected into K562 cells. A stable transfectant, known as pXJ41-asELDF15, stably produced antisense ELDF15. Compared with K562 and K562-zeo cells, K562-pXJ41-asELDF15 cells showed inhibition of cell proliferation. RT-PCR analysis showed that the expression and protein level of ELDF15 decreased significantly in K562 cells transfected with pXJ41-asELDF15. Expression of hemoglobin increased in K562 cells transfected with pXJ41-asELDF15 by benzidine staining. increases NBT reduction activity in K562 cells transfected with pXJ41-asELDF15.Colony forming efficiency in two-layer soft agar was clearly inhibited as assessed by electron microscopy. These results suggest that ELDF15 plays a potential role in cell differentiation, proliferation and carcinogenesis.

• BMB Reports
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• v.46 no.1
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• pp.59-64
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• 2013

Signal transducer and activator of transcription 3 (STAT3) and telomerase are considered attractive targets for anticancer therapy. The in vitro anticancer activity of the gold(I) compound auranofin was investigated using MDA-MB 231 human breast cancer cells, in which STAT3 is constitutively active. In cell culture, auranofin inhibited growth in a dose-dependent manner, and N-acetyl-L-cysteine (NAC), a scavenger of reactive oxygen species (ROS), markedly blocked the effect of auranofin. Incorporation of 5-bromo-2'-deoxyuridine into DNA and anchorage-independent cell growth on soft agar were decreased by auranofin treatment. STAT3 phosphorylation and telomerase activity were also attenuated in cells exposed to auranofin, but NAC pretreatment restored STAT3 phosphorylation and telomerase activity in these cells. These findings indicate that auranofin exerts in vitro antitumor effects in MDA-MB 231 cells and its activity involves inhibition of STAT3 and telomerase. Thus, auranofin shows potential as a novel anticancer drug that targets STAT3 and telomerase.