• Title/Summary/Keyword: Colony characteristics

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Carcinogenic Potentials of HPV-16 and NNK in Human in Vitro Model (인체 세포 모델을 이용한 HPV-16과 NNK의 발암 잠재력에 관한 연구)

  • 양재호;이세영
    • Toxicological Research
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    • v.12 no.2
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    • pp.271-275
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    • 1996
  • Carcinogenic potential of HPV-16 DNA and NNK in a human keratinocyte cell line was assessed to study effects of viral-chemical interaction. Human cells were transfected with HPV-16 DNA and 6 clonal cell lines were subsequently obtained. Clonal line-3 and 6 at passage 7 showed characteristics of tumor cells such as increases of saturation density, soft-agar colony formation, cell aggregation and foci appearance. Among cells treated with 1$\mu M$, 10$\mu M$, 100$\mu M$ or 1 mM of NNK for 4 weeks, 100$\mu M$ treatment showed most tumorigenic characteristics at passage 7. These results indicate that either HPV-16 or NNK alone is tumorigenic in this in human in vitro model. When cells transfected with HPV-16 were subsequently exposed by 100 uM NNK for 4 weeks, all the clonal cells except clone-1 showed higher levels of tumor cell characteristics than HPV-16 DNA or NNK exposure alone. Clonal line-6, the most tumorigenic cells, showed higher transcriptional level of fibronectin and lower level of TGF-$\beta_1$, as compared to control cells, suggesting that alteration of growth factor or extracellular matrix may play a role in carcinogenesis process induced by HPV-16 and NNK. Taken together, the present study indicates that viral-chemical interactions between HPV-16 DNA and NNK enhance carcinogenic potentials of human cells and implies that smoking among people infected with human papillomavirus may pose an additional risk of causing cancer.

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Inhibition of growth and biofilm formation of Staphylococcus aureus by corosolic acid (Corosolic acid에 의한 Staphylococcus aureus의 생장 및 생물막 형성 저해)

  • Yum, Su-Jin;Kim, Seung Min;Yu, Yeon-Cheol;Jeong, Hee Gon
    • Korean Journal of Food Science and Technology
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    • v.49 no.2
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    • pp.146-150
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    • 2017
  • Staphylococcus aureus is a pathogenic bacterium that causes food poisoning, exhibits a strong capacity to form biofilm, and is highly resistant to antimicrobial agents. The purpose of this study was to investigate the antimicrobial characteristics of corosolic acid against S. aureus. S. aureus showed high susceptibility to corosolic acid in a concentration-dependent manner. The minimum inhibitory concentration and colony-forming ability determined by the broth microdilution method showed that corosolic acid had strong antimicrobial activity against the bacteria. The diameters of the inhibition zone and numbers of colony forming units at each concentration of corosolic acid were also measured. In addition, corosolic acid displayed potent biofilm inhibition activity against S. aureus at concentrations below its minimum inhibitory concentration. These results suggest that corosolic acid can be used to effectively prevent biofilm formation by S. aureus, thereby making S. aureus more susceptible to the action of antimicrobials.

Inhibitory activities of Perilla frutescens britton leaf extract against the growth, migration, and adhesion of human cancer cells

  • Kwak, Youngeun;Ju, Jihyeung
    • Nutrition Research and Practice
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    • v.9 no.1
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    • pp.11-16
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    • 2015
  • BACKGROUND/OBJECTIVES: Perilla frutescens Britton leaves are a commonly consumed vegetable in different Asian countries including Korea. Cancer is a major cause of human death worldwide. The aim of the current study was to investigate the inhibitory effects of ethanol extract of perilla leaf (PLE) against important characteristics of cancer cells, including unrestricted growth, resisted apoptosis, and activated metastasis, using human cancer cells. MATERIALS/METHODS: Two human cancer cell lines were used in this study, HCT116 colorectal carcinoma cells and H1299 non-small cell lung carcinoma cells. Assays using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide were performed for measurement of cell growth. Soft agar and wound healing assays were performed to determine colony formation and cell migration, respectively. Nuclear staining and cell cycle analysis were performed for assessment of apoptosis. Fibronectin-coated plates were used to determine cell adhesion. RESULTS: Treatment of HCT116 and H1299 cells with PLE resulted in dose-dependent inhibition of growth by 52-92% (at the concentrations of 87.5, 175, and $350{\mu}g/ml$) and completely abolished the colony formation in soft agar (at the concentration of $350{\mu}g/ml$). Treatment with PLE at the $350{\mu}g/ml$ concentration resulted in change of the nucleus morphology and significantly increased sub-G1 cell population in both cells, indicating its apoptosis-inducing activity. PLE at the concentration range of 87.5 to $350{\mu}g/ml$ was also effective in inhibiting the migration of H1299 cells (by 52-58%) and adhesion of both HCT116 and H1299 cells (by 25-46%). CONCLUSIONS: These results indicate that PLE exerts anti-cancer activities against colon and lung cancers in vitro. Further studies are needed in order to determine whether similar effects are reproduced in vivo.

Evaluation of the predisposing factors and involved outcome of surgical treatment in bisphosphonate-related osteonecrosis of the jaw cases including bone biopsies

  • Kim, Tae-Hwan;Seo, Won-Gyo;Koo, Chul-Hong;Lee, Jae-Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.42 no.4
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    • pp.193-204
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    • 2016
  • Objectives: This study examined the statistical relevance of whether the systemic predisposing factors affect the prognosis of surgical treatment of bisphosphonate-related osteonecrosis of the jaw (BRONJ). All cases had undergone bone biopsies to determine the characteristics of the mechanisms of BRONJ by optical microscopy. Materials and Methods: The data included 54 BRONJ cases who underwent surgery and in whom bone biopsies were performed. The results of surgery were evaluated and the results were classified into 3 categories: normal recovery, delayed recovery, and recurrence after surgery. The medical history, such as diabetes mellitus, medication of steroids, malignancies on other sites was investigated for an evaluation of the systemic predisposing factors in relation to the prognosis. The three factors involved with the medication of bisphosphonate (BP) were the medication route, medication period, and drug holiday of BP before surgery. The serum C-terminal cross-linking telopeptide (CTX) value and presence of microorganism colony in bone biopsy specimens were also checked. Statistical analysis was then carried out to determine the relationship between these factors and the results of surgery. Results: The group of patients suffering from diabetes and on steroids tended to show poorer results after surgery. Parenteral medication of BP made the patients have a poorer prognosis after surgery than oral medication. In contrast, the medication period and drug holiday of BP before surgery did not have significance with the results of surgery nor did the serum CTX value and presence of microorganism colony. Necrotic bone specimens in this study typically showed disappearing new bone formation around the osteocytic lacunae and destroyed Howship's lacunae. Conclusion: Although many variables exist, this study could in part, predict the prognosis of surgical treatment of BRONJ by taking the patient's medical history.

Screening of Cholesterol-lowering Bifidobacterium from Guizhou Xiang Pigs, and Evaluation of Its Tolerance to Oxygen, Acid, and Bile

  • Zhang, Rujiao;He, Laping;Zhang, Ling;Li, Cuiqin;Zhu, Qiujin
    • Food Science of Animal Resources
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    • v.36 no.1
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    • pp.37-43
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    • 2016
  • Cardiovascular and cerebrovascular diseases seriously harm human health, and Bifidobacterium is the most beneficial probiotic in the gastrointestinal tract of humans. This work aimed to screen cholesterol-lowering Bifidobacterium from Guizhou Xiang Pig and evaluate its tolerance to oxygen, acid, and bile. Twenty-seven aerotolerant strains with similar colony to Bifidobacterium were isolated through incubation at 37℃ in 20% (v/v) CO2-80% (v/v) atmospheric air by using Mupirocin lithium modified MRS agar medium, modified PTYG with added CaCO3, and modified PTYG supplemented with X-gal. Ten strains with cholesterol-lowering rates above 20% (w/w) were used for further screening. The selected strains’ tolerance to acid and bile was then determined. A combination of colony and cell morphology, physiological, and biochemical experiments, as well as 16S rRNA gene-sequence analysis, was performed. Results suggested that BZ25 with excellent characteristics of high cholesterol-removal rate of 36.32% (w/w), as well as tolerance to acid and bile, was identified as Bifidobacterium animalis subsp. lactis. To further evaluate Bifidobacterium BZ25’s growth characteristic and tolerance to oxygen, culture experiments were performed in liquid medium and an agar plate. Findings suggested that BZ25 grew well both in environmental 20% (v/v) CO2-80% (v/v) atmospheric air and in 100% atmospheric air because BZ25 reached an absorbance of 1.185 at 600 nm in 100% atmospheric air. Moreover, BZ25 was aerotolerant and can grow in an agar medium under the environmental condition of 100% atmospheric air. This study can lay a preliminary foundation for the potential industrial applications of BZ25.

Identification of a Pathogenic Bacterium, Staphylococcus gallinarum, to Bombyx mori (누에(Bombyx mori) 무름병을 야기하는 병원성 세균 Staphylococcus gallinarum의 동정)

  • 김길호;박영진;김용균
    • Korean journal of applied entomology
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    • v.41 no.4
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    • pp.279-284
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    • 2002
  • Flacherie symptom was found in the fifth instar larvae of silkworm, Bombyx mori. The bacterial pathogen was isolated from the hemolymph of the infected silkworm and identified. The isolated bacteria caused a significant flacherie pathogenicity to the fifth instar larvae of B. mori when $5{\times}10^{6}$ cfu (colony-forming unit) of the bacteria was injected into each larva. The infected larvae began to die at 6 days after injection and resulted in complete mortality at 10 days. The bacterium was identified as Staphylococcus gallinarum based on the morphological and physiological characteristics described in Bergey's manual. This identification was further supported by the characters of carbohydrate utility analyzed from a bacterial identification system ($MicroLog^{\circledR}$) and also by the molecular structure of 165-23S rDNA internal transcribed spaces. As an insecticidal action, S. gallinarum caused hemolymph septicemia by its cytotoxic effect on the hemocytes of B. mori.

Characteristics of High Acidity Producing Acetic Acid Bacteria Isolated from Industrial Vinegar Fermentation (발효 중인 양초식초에서 분리한 고산도 초산균의 생육특성)

  • Park, Mi-Hwa;Lyu, Dong-Kyu;Ryu, Chung-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.3
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    • pp.394-398
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    • 2002
  • Acetic acid bacteria have been isolated from running high-acid vinegar fermentation. The color of the isolated colony was beige-yellowish. Isolated cell was rod-shaped, small, pale, absolutely aerobic and gram-negative. Microscopically the cells appeared as non-motile and non-flagellated, preferentially occuring in pairs. The optimum temperature and pH for culture were 30$\^{C}$ and 2.7, respectively. The strain was able to grow in the presence of acetic acid, ethanol and glucose. Ethanol was oxidized to acetic acid which was not oxidized any more. The isolated strain utilized glucose, fructose, maltose, sucrose, mannitol and sorbitol as carbon source. Cellulose formation was not detected on bouillon. The DNA (G +C) content of isolated strain was determined to be 56.2 mol%. The strain isolated from industrial vinegar fermentation was identified as Gluconacetobacter europaeus.

Characteristics of Cyanobacteria and Odorous Compounds Production in Lake Uiam and Lower Gonji Stream (의암호와 공지천 하류에서 남조류와 냄새물질의 발생 특징)

  • Youn, Seok Jea;Im, Jong Kwon;Byeon, Myeong-Seop;Yu, Soon Ju
    • Journal of Korean Society on Water Environment
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    • v.35 no.2
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    • pp.99-104
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    • 2019
  • The objective of this study was to investigate the relationship between the biomass of cyanobacteria and the concentration of 2-methylisoborneol (2-MIB) in the lower Gongji stream. The investigation was done using a field study that was conducted from 2015 to 2017. The 2-MIB concentration in the lower Gongji stream ranged from 0 to 153 ng/L, while the upper stream had 0 2-MIB concentration. 3 genera (Oscillatoria, Phormidium, Pseudanabaena) of cyanobacteria were detected in the lower Gongji stream with 2-MIB concentration. Among these 3 genera, an increase in Phormidium, Pseudanabaena biomass was associated with an increase in 2-MIB concentration. Accordingly, Phormidium, Pseudanabaena were regarded as the biological source of 2-MIB in that area. In October 2017, although planktonic cyanobacteria occurred less frequently, many benthic cyanobacteria mats were observed on the surface of the water body. Therefore, the high 2-MIB concentration, which exceeded 110 ng/L, can likely be attributed to the benthic cyanobacteria. In a laboratory experiment, individual Oscillatoria filaments were aggregated to form a colony with a higher density. This colony tended to float on the water surface. Cyanobacteria mats after floating aggregated mats were distributed in a net shape on the bottom.

Antibacterial activity of enrofloxacin loaded gelatin-sodium alginate composite nanogels against intracellular Staphylococcus aureus small colony variants

  • Luo, Wanhe;Liu, Jinhuan;Algharib, Samah Attia;Chen, Wei
    • Journal of Veterinary Science
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    • v.23 no.3
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    • pp.48.1-48.12
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    • 2022
  • Background: The poor intracellular concentration of enrofloxacin might lead to treatment failure of cow mastitis caused by Staphylococcus aureus small colony variants (SASCVs). Objectives: In this study, enrofloxacin composite nanogels were developed to increase the intracellular therapeutic drug concentrations and enhance the efficacy of enrofloxacin against cow mastitis caused by intracellular SASCVs. Methods: Enrofloxacin composite nanogels were formulated by an electrostatic interaction between gelatin (positive charge) and sodium alginate (SA; negative charge) with the help of CaCl2 (ionic crosslinkers) and optimized by a single factor test using the particle diameter, zeta potential (ZP), polydispersity index (PDI), loading capacity (LC), and encapsulation efficiency (EE) as indexes. The formation mechanism, structural characteristics, bioadhesion ability, cellular uptake, and the antibacterial activity of the enrofloxacin composite nanogels against intracellular SASCVs strain were studied systematically. Results: The optimized formulation was comprised of 10 mg/mL (gelatin), 5 mg/mL (SA), and 0.25 mg/mL (CaCl2). The size, LC, EE, PDI, and ZP of the optimized enrofloxacin composite nanogels were 323.2 ± 4.3 nm, 15.4% ± 0.2%, 69.6% ± 1.3%, 0.11 ± 0.02, and -34.4 ± 0.8 mV, respectively. Transmission electron microscopy showed that the enrofloxacin composite nanogels were spherical with a smooth surface and good particle size distributions. In addition, the enrofloxacin composite nanogels could enhance the bioadhesion capacity of enrofloxacin for the SASCVs strain by adhesive studies. The minimum inhibitory concentration, minimum bactericidal concentration, minimum biofilm inhibitory concentration, and minimum biofilm eradication concentration were 2, 4, 4, and 8 ㎍/mL, respectively. The killing rate curve had a concentration-dependent bactericidal effect as increasing drug concentrations induced swifter and more radical killing effects. Conclusions: This study provides a good tendency for developing enrofloxacin composite nanogels for treating cow mastitis caused by intracellular SASCVs and other intracellular bacterial infections.

Enhanced antibacterial activity of tilmicosin against Staphylococcus aureus small colony variants by chitosan oligosaccharide-sodium carboxymethyl cellulose composite nanogels

  • Luo, Wanhe;Liu, Jinhuan;Zhang, Shanling;Song, Wei;Algharib, Samah Attia;Chen, Wei
    • Journal of Veterinary Science
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    • v.23 no.1
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    • pp.1.1-1.11
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    • 2022
  • Background: The poor bioadhesion capacity of tilmicosin resulting in treatment failure for Staphylococcus aureus small colony variants (SASCVs) mastitis. Objectives: This study aimed to increase the bioadhesion capacity of tilmicosin for the SASCVs strain and improve the antibacterial effect of tilmicosin against cow mastitis caused by the SASCVs strain. Methods: Tilmicosin-loaded chitosan oligosaccharide (COS)-sodium carboxymethyl cellulose (CMC) composite nanogels were formulated by an electrostatic interaction between COS (positive charge) and CMC (negative charge) using sodium tripolyphosphate (TPP) (ionic crosslinkers). The formation mechanism, structural characteristics, bioadhesion, and antibacterial activity of tilmicosin composite nanogels were studied systematically. Results: The optimized formulation was comprised of 50 mg/mL (COS), 32 mg/mL (CMC), and 0.25 mg/mL (TPP). The size, encapsulation efficiency, loading capacity, polydispersity index, and zeta potential of the optimized tilmicosin composite nanogels were 357.4 ± 2.6 nm, 65.4 ± 0.4%, 21.9 ± 0.4%, 0.11 ± 0.01, and -37.1 ± 0.4 mV, respectively; the sedimentation rate was one. Scanning electron microscopy showed that tilmicosin might be incorporated in nano-sized crosslinked polymeric networks. Moreover, adhesive studies suggested that tilmicosin composite nanogels could enhance the bioadhesion capacity of tilmicosin for the SASCVs strain. The inhibition zone of native tilmicosin, tilmicosin standard, and tilmicosin composite nanogels were 2.13 ± 0.07, 3.35 ± 0.11, and 1.46 ± 0.04 cm, respectively. The minimum inhibitory concentration of native tilmicosin, tilmicosin standard, and tilmicosin composite nanogels against the SASCVs strain were 2, 1, and 1 ㎍/mL, respectively. The in vitro time-killing curves showed that the tilmicosin composite nanogels increased the antibacterial activity against the SASCVs strain. Conclusions: This study provides a potential strategy for developing tilmicosin composite nanogels to treat cow mastitis caused by the SASCVs strain.