• Journal of Microbiology and Biotechnology
• /
• v.28 no.1
• /
• pp.77-86
• /
• 2018

The human pathogen and food spoiler Bacillus cereus can form biofilms that act as a persistent source of contamination, which is of public health concern. This study aimed to understand how the source of isolation might affect the behavior of biofilm formation. Biofilm formation abilities of 56 strains of B. cereus isolated from different environments, including human food poisoning, farm, and food, were determined. Crystal violet assay results revealed significant (p < 0.05) differences in biofilm formation abilities among the strains isolated from different sources only at an early stage of incubation. However, strain origin showed no impact on later stage of biofilm formation. Next, correlation of the group of isolates on the basis of their biofilm-forming abilities with the number of sessile cells, sporulation, and extracellular polymeric substance (EPS) formation was determined. The number of sessile cells and spores in biofilms was greatly influenced by the groups of isolates that formed dense, moderate, and weak biofilms. The contribution of extracellular DNA and/or proteins to EPS formation was also positively correlated with biofilm formation abilities. Our results that the source of isolation had significant impact on biofilm formation might provide important information to develop strategies to control B. cereus biofilm formation.

• The Journal of Korean Medicine
• /
• v.21 no.4
• /
• pp.183-192
• /
• 2000

Objectives : The present study was carried out to investigate the effects of Danchun-hwan(DCH) on the peroxynitrite-induced neural cell death in human neuroblastoma cell line, SH-SY5Y. Methods : The cultured cells were pretreated with DCH and exposed to 3-morpholinosydnonimine(SIN-1) that simultaneously generates NO and superoxide, thus possibly forming peroxynitrite. The cell damage was assessed by using MTT assay and crystal violet staining. Results : Exposure of the cells to SIN-1 for 24hr induced 75% apoptotic cell death, as evaluated by the occurrence of morphological nuclear changes characteristic of apoptosis using 4', 6-diamidino-2-phenylinole(DAPI). However, pretreatment of SH-SY5Y with the water extracts of DCH, inhibited the apoptotic cell death in a dose-dependent manner. DCH also inhibited SIN-1-induced apoptotic caspase 3-like protease activity in a dose-dependent manner. DCH recovered the depleted glutathione levels by SIN-1. Conclusions : Taken together, it is suggested that DCH protected human neuroblastoma cell line, SH-SY5Y, from the free radical injury mediated by peroxynitrite by a mechanism of elevating antioxidant, GSH.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.53 no.6
• /
• pp.878-885
• /
• 2020

Previously, we reported that globefish Takifugu obscurus homogenate suppresses the growth of human colorectal cancer cells. To extend the applications of globefish homogenate, we investigated its cytotoxic effects on human breast cancer cells. To assess the effects of globefish homogenate on growth of MCF (Michigan Cancer Foundation)-7 human breast cancer cells, cell proliferation and colony formation assays were performed using the cell counting and Crystal Violet staining methods. The 50% inhibitory concentration (IC50) of globefish homogenate on MCF-7 cell proliferation was calculated from the sigmoidal dose-response curve. The colony formation assay demonstrated that MCF-7 cells treated with globefish homogenate formed up to 80% fewer colonies than control MCF-7 cells. Treatment with globefish homogenate markedly suppressed the growth of MCF-7 cells in a dose-dependent manner. The sensitivity of the cells to globefish homogenate was determined by calculating the IC50; in this case, the IC50 was 210 ㎍/mL. Furthermore, significant downregulation of Cyclin D1 expression, along with phospho-Akt and total Akt levels, was observed in MCF-7 cells treated with globefish homogenate. This study demonstrates that treatment with globefish homogenate inhibits the proliferation of MCF-7 human breast cancer cells by downregulating the expression of phosphor-Akt, total Akt, and Cyclin D1 proteins.

• Journal of Microbiology and Biotechnology
• /
• v.33 no.6
• /
• pp.715-723
• /
• 2023

Microbial biofilms are resilient, immune-evasive, often antibiotic-resistant health challenges, and increasingly the target for research into novel therapeutic strategies. We evaluated the effects of a nutraceutical enzyme and botanical blend (NEBB) on established biofilm. Five microbial strains with known implications in chronic human illnesses were tested: Candida albicans, Staphylococcus aureus, Staphylococcus simulans (coagulase-negative, penicillin-resistant), Borrelia burgdorferi, and Pseudomonas aeruginosa. The strains were allowed to form biofilm in vitro. Biofilm cultures were treated with NEBB containing enzymes targeted at lipids, proteins, and sugars, also containing the mucolytic compound N-acetyl cysteine, along with antimicrobial extracts from cranberry, berberine, rosemary, and peppermint. The post-treatment biofilm mass was evaluated by crystal-violet staining, and metabolic activity was measured using the MTT assay. Average biofilm mass and metabolic activity for NEBB-treated biofilms were compared to the average of untreated control cultures. Treatment of established biofilm with NEBB resulted in biofilm-disruption, involving significant reductions in biofilm mass and metabolic activity for Candida and both Staphylococcus species. For B. burgdorferi, we observed reduced biofilm mass, but the remaining residual biofilm showed a mild increase in metabolic activity, suggesting a shift from metabolically quiescent, treatment-resistant persister forms of B. burgdorferi to a more active form, potentially more recognizable by the host immune system. For P. aeruginosa, low doses of NEBB significantly reduced biofilm mass and metabolic activity while higher doses of NEBB increased biofilm mass and metabolic activity. The results suggest that targeted nutraceutical support may help disrupt biofilm communities, offering new facets for integrative combinational treatment strategies.

• Korean Journal of Microbiology
• /
• v.50 no.4
• /
• pp.360-367
• /
• 2014

Candida albicans is an opportunistic and the most prevalent fungal pathogen that can cause superficial and systemic infections in immunocompromised patients. C. albicans can promote the transition from budding yeast to filamentous form, generating biofilms. Infections associated with C. albicans biofilms are frequently resistant to conventional antifungal therapy. Therefore, the development of more effective antifungal drugs related with biofilm formation is required urgently. The roots of Rheum undulatum have been used for medicinal purposes in Korea and China traditionally. The aim of present study was to evaluate the effect of R. undulatum extract upon preformed biofilms of 12 clinical C. albicans isolates and the antifungal activities. Its effect on preformed biofilms was evaluated using XTT reduction assay, and metabolic activity of all tested strains was reduced significantly ($49.4{\pm}6.0%$) at 0.098 mg/ml R. undulatum. The R. undulatum extract blocked the adhesion of C. albicans biofilms to polystyrene surfaces, and damaged the cell membrane integrity of C. albicans which was analyzed by CFDA, AM, and propidium iodide double staining. It caused cell lysis which was observed by Confocal laser scanning and phase contrast microscope after propidium iodide and neutral red staining, respectively. Membrane permeability was changed as evidenced by crystal violet uptake. The data suggest that R. undulatum inhibits biofilm formation by C. albicans, which can be associated with the damage of the cell membrane integrity, the changes in the membrane permeability and the cell lysis of C. albicans.

• The Korean Journal of Mycology
• /
• v.26 no.1 s.84
• /
• pp.8-15
• /
• 1998

The present research was undertaken to investigate the activity of ligninolytic enzymes and the decolorization capability of some dyes with Irpex zonatus BN2, isolated from nature and identified. For the assay of enzyme activities, the isolate did not produce lignin peroxidase (LiP) and veratryl alcohol oxidase (VAO), but laccase and manganese dependent peroxidase (MnP). While the activity for MnP was low $(61.6\;nmol/mg{\cdot}protein)$, its laccase activity was very high $(1185.9\;nmol/mg{\cdot}protein)$. Moreover, laccase had appeared earlier than MnP. When the isolate was incubated with each dye for 10 days, the decolorization rates of azo dyes, such as orange II, orange G, tropaeolin O and congo red were 98.0%, 97.4%, 99.0% and 95.3%, respectively. In case of heterocyclic dyes, eosin Y, toludine blue, methyl blue and azur B were 97.4 %, 98.7%, 99.9% and 94.0% respectively. Finally the results of triphenylmethane dye such as basic fuchsin, malachite green and crystal violet were 98.5%, 95.7% and 99.4%, respectively. The results suggest that laccase of Irpex zonatus BN2 should be played an important role in the decolorization of the dyes.

• Journal of Korean society of Dental Hygiene
• /
• v.21 no.5
• /
• pp.527-533
• /
• 2021

Objectives: Electrolyzed water has been proven to have antibacterial effects against various microorganisms. However, there are only a few studies about effects of electrolyzed water on oral bacteria. The purpose of this study was to examine the antibacterial effect of electrolyzed water on Streptococcus mutans in vitro. Methods: S. mutans KCOM 1054 was treated with electrolyzed water for 1 or 3 minutes and plated on Mitis Salivarius agar with 15% sucrose and bacitracin. After incubation for 48 hours, colony forming units (CFU) were counted, and dental plaque was quantified by crystal violet staining. Results: The growth of S. mutans was significantly inhibited by electrolyzed water (p<0.001). In addition, the dental plaque formation by S. mutans was decreased in a time-dependent manner by exposure to electrolyzed water (p<0.001). Conclusions: Our results suggest that electrolyzed water can effectively prevent dental caries by inhibiting growth of (and the formation of dental plaque by) S. mutans.

• Korean Journal of Clinical Laboratory Science
• /
• v.51 no.3
• /
• pp.379-385
• /
• 2019

Osteoporosis is a disease that increases the risk of fractures by inducing a decrease in bone strength by the changes in hormones and a decrease in minerals. Recent reports have indicated that the long-term administration of Adefovir dipivoxil (ADV), which is used as a treatment for the hepatitis virus and AIDS, may have osteoporotic side effects. On the other hand, there are few studies on the cytopathic correlation of these causes. In this study, the biological relevance of ADV was evaluated using osteoblast hFOB1.19 and vascular endothelial cell HUVEC. First, the cells were treated with ADV at different concentrations, and DAPI and crystal violet staining were performed for morphological analysis of each cell and nucleus. A CCK-8 assay, real-time PCR, alkaline phosphatase (ALP) staining, and activity was performed to evaluate the drug effects on cell proliferation, gene expression, and osteoblast differentiation. As a result, ADV induced cell hypertrophy in hFOB1.19 cells and HUVEC cells. Furthermore, ADV not only inhibited cell proliferation and TGF-${\beta}$ expression but was also involved in osteoblast differentiation. Overall, these results provide basic data to help better understand the mechanism of ADV-induced osteoporosis and its clinical implications.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.9
• /
• pp.1216-1225
• /
• 2014

Biofilm-related infections of Candida albicans are a frequent cause of morbidity and mortality in hospitalized patients, especially those with immunocompromised status. Options of the antifungal drugs available for successful treatment of drug-resistant biofilms are very few, and as such, new strategies need to be explored against them. The aim of this study was to evaluate the efficacy of phenylpropanoids of plant origin against planktonic cells, important virulence factors, and biofilm forms of C. albicans. Standard susceptibility testing protocol was used to evaluate the activities of 13 phenylpropanoids against planktonic growth. Their effects on adhesion and yeast-to-hyphae morphogenesis were studied in microplate-based methodologies. An in vitro biofilm model analyzed the phenylpropanoid-mediated prevention of biofilm development and mature biofilms using XTT-metabolic assay, crystal violet assay, and light microscopy. Six molecules exhibited fungistatic activity at ${\leq}0.5mg/ml$, of which four were fungicidal at low concentrations. Seven phenylpropanoids inhibited yeast-to-hyphae transition at low concentrations (0.031-0.5 mg/ml), whereas adhesion to the solid substrate was prevented in the range of 0.5-2 mg/ml. Treatment with ${\leq}0.5mg/ml$ concentrations of at least six small molecules resulted in significant (p < 0.05) inhibition of biofilm formation by C. albicans. Mature biofilms that are highly resistant to antifungal drugs were susceptible to low concentrations of 4 of the 13 molecules. This study revealed phenylpropanoids of plant origin as promising candidates to devise preventive strategies against drug-resistant biofilms of C. albicans.

• Journal of Microbiology and Biotechnology
• /
• v.32 no.2
• /
• pp.263-267
• /
• 2022

The aim of this study was to determine whether the antibacterial activity of chitosan-modified Fe₃O₄ (CS@Fe₃O₄) nanomaterials against Acinetobacter baumannii (A. baumannii) is mediated through changes in biofilm formation and reactive oxygen species (ROS) production. For this purpose, the broth dilution method was used to examine the effect of CS@Fe₃O₄ nanoparticles on bacterial growth. The effects of CS@Fe₃O₄ nanoparticles on biofilm formation were measured using a semi-quantitative crystal violet staining assay. In addition, a bacterial ROS detection kit was used to detect the production of ROS in bacteria. The results showed that CS@Fe₃O₄ nanoparticles had a significant inhibitory effect on the colony growth and biofilm formation of drug-resistant A. baumannii (p < 0.05). The ROS stress assay revealed significantly higher ROS levels in A. baumannii subjected to CS@Fe₃O₄ nanoparticle treatment than the control group (p < 0.05). Thus, we demonstrated for the first time that CS@Fe₃O₄ nanoparticles had an inhibitory effect on A. baumannii in vitro, and that the antibacterial effect of CS@Fe₃O₄ nanoparticles on drug-resistant A. baumannii was more significant than on drug-sensitive bacteria. Our findings suggest that the antibacterial mechanism of CS@Fe₃O₄ nanoparticles is mediated through inhibition of biofilm formation in drug-resistant bacteria, as well as stimulation of A. baumannii to produce ROS. In summary, our data indicate that CS@Fe₃O₄ nanoparticles could be used to treat infections caused by drug-resistant A. baumannii.