• Title/Summary/Keyword: Anti-bacterial study

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Effect of 1-[(2-Hydroxyethoxy)methyl]-6-(phenylthio)thymine (HEPT) Derivatives on Bacterial Growth

  • Gang, Jin-Gu;Yun, Hong-Chul;Son, Jong-Chan;Hwang, Se-Young
    • Journal of Microbiology and Biotechnology
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    • v.8 no.3
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    • pp.229-236
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    • 1998
  • 6-Substituted derivatives of 1-[(2-hydroxyethoxy)methyl]-6-(phenylthio)thymine (HEPT) were synthesized by introducing alkyl groups with the aid of chlorotrimethylsilane, and then purified ranging 40 to 81 % of yield. Because of their peculiar structures, we presumed that HEPT derivatives would contain extra biological activities other than their already known anti-human immunodeficiency viral (HIV -1) activities. In this study, we investigated the possible effects of the HEPT derivatives on bacterial growth and found their selective antibiotic activities against gram-positive strains. We could not observe the corresponding activity from a disc-zone test, but confirmed the activity by liquid cultivation. Since the growth rate of cells was easily recovered, the antibiotic function was suggested to be bacteriostatic. We also suggested that the intracellular fate of HEPT derivatives would be fast. A HEPT derivative f-3 was shown to synergize unidirectionally toward chloramphenicol (Chr). With 0.1 mM f-3, the Chr-directed growth-inhibitory curve appeared 4 hours earlier than found without the additive. Interestingly, from the data of SDS-polyacrylamide gel electrophoresis (PAGE), we found that a membrane-bound protein having a molecular weight of 70-kDa was overexpressed by f-3 in S. aureus.

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Subeschar culture using a punch instrument in unstageable wounds

  • Jung, Han Byul;Lee, Yong Jig
    • Archives of Plastic Surgery
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    • v.47 no.3
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    • pp.228-234
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    • 2020
  • Background A patient's overall condition sometimes does not allow for the complete removal of a dead eschar or injured slough in cases involving a pressure-injury skin lesion. This frequently occurs in clinical practice, particularly in bedridden and older patients receiving home care or intensive care. Even after debridement, it is also difficult to manage open exudative wounds in these patients. Nevertheless, when a mature or immature eschar is treated without proper debridement, liquefaction necrosis underneath the eschar or slough tends to reveal a large, open wound with infectious exudates. We hypothesized that if the presence of any bacteria under the eschar can be evaluated and the progression of the presumed infection of the subeschar can be halted or delayed without creating an open wound, the final wound can be small, shallow, and uninfected. Methods Using a punch instrument, we performed 34 viable subeschar tissue cultures with a secure junction between the eschar and the normal skin. Results The bacterial study had 29 positive results. Based on these results and the patient's status, appropriate antibiotics could be selected and administered. The use of suitable antibiotics led to relatively shallow and small exposed wounds. Conclusions This procedure could be used to detect potentially pathogenic bacteria hidden under black or yellow eschars. Since subeschar infections are often accompanied by multidrug-resistant bacteria, the early detection of hidden infections and the use of appropriate antibiotics are expected to be helpful to patients.

Perturbation of host responses by Porphyromonas gingivalis biofilm (Porphyromonas gingivalis 바이오필름에 의한 숙주 면역반응의 교란)

  • Jeon, Woo-Seok;Kim, Sung-Jo;Choi, Jeom-Il
    • Journal of Periodontal and Implant Science
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    • v.32 no.4
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    • pp.827-836
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    • 2002
  • The present study was performed to evaluate how cellular and humoral immune responses were perturbed by immunization of mixed periodontal bacterial biofilms. Each group of mice was immunizared with 1) Poqhyromonas gingivalis (P. gingivaliis) grown as a planktonic culture, 2) Fusobacterium nucleatum (F. nucleatum), 3) P. gingivalis grown as a biofilm, or 4) mixed P. gingivalis plus F. nucleatum grown as a biofilm culture, respectively. Immune mouse sera were collected from each mouse. Spleens were harvested to isolate T cells and consequently stimulated with antigen presenting cells and P. gingivalis whole cell antigen to establish P. gingivalis-specific T cell lines. There were no significant differences in the mean anti- gingivalis IgG antibody titers among mouse groups. Immunization of mice with pure P. gingivalis biofilm or mixed P gingivalis plus F. nucleatum biofilm resulted in significant reduction o f antibody avidity and opsonophagocytois function. INF-$\gamma$production by P. gingivalis-specific T cell lines was also substantially recluced in mouse groups immunized with the biofilm. It was concluded that P. gingivalis biofilm perturbs the cellular and humoral immune responses in periodontal disease.

Proteomic and Phenotypic Analyses of a Putative YggS Family Pyridoxal Phosphate-Dependent Enzyme in Acidovorax citrulli

  • Lynn Heo;Yongmin Cho;Junhyeok Choi;Jeongwook Lee;Yoobin Han;Sang-Wook Han
    • The Plant Pathology Journal
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    • v.39 no.3
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    • pp.235-244
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    • 2023
  • Acidovorax citrulli (Ac) is a phytopathogenic bacterium that causes bacterial fruit blotch (BFB) in cucurbit crops, including watermelon. However, there are no effective methods to control this disease. YggS family pyridoxal phosphate-dependent enzyme acts as a coenzyme in all transamination reactions, but its function in Ac is poorly understood. Therefore, this study uses proteomic and phenotypic analyses to characterize the functions. The Ac strain lacking the YggS family pyridoxal phosphate-dependent enzyme, AcΔyppAc(EV), virulence was wholly eradicated in geminated seed inoculation and leaf infiltration. AcΔyppAc(EV) propagation was inhibited when exposed to L-homoserine but not pyridoxine. Wild-type and mutant growth were comparable in the liquid media but not in the solid media in the minimal condition. The comparative proteomic analysis revealed that YppAc is primarily involved in cell motility and wall/membrane/envelop biogenesis. In addition, AcΔyppAc(EV) reduced biofilm formation and twitching halo production, indicating that YppAc is involved in various cellular mechanisms and possesses pleiotropic effects. Therefore, this identified protein is a potential target for developing an efficient anti-virulence reagent to control BFB.

Immune Response of Bacterial Proteins of Staphylococcus intermedius from Canine Atopic Dermatitis (개의 아토피성 피부염에서 분리한 Staphylococcus intermedius 균의 세균단백질의 면역반응)

  • Park, Hee-myung
    • Journal of Veterinary Clinics
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    • v.21 no.1
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    • pp.20-22
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    • 2004
  • Bacterial infection of canine atopic dermatitis is largely caused by Staphylococcus intermedius and may be a superficial or deep pyoderma. The Purpose of this study was to identify the major proteins of S. intermedius cell surface components in humoral immune response of atopic dermatitis dog. Sera samples were obtained from dogs with atopic dermatitis and superficial pyoderma referred to the Veterinary Medical Teaching Hospital, Konkuk University. An isolate of S. intermedius from a clinical case of canine atopic dermatitis was cultured in brain heart infusion broth overnight at $37^{\circ}C$ in aerobic conditions on an orbital shaker. Following culture, Staphylococci were harvested by centrifugation, washed in PBS, and resuspended in PBS containing lysostaphin. The soluble components were separated by centrifugation and were collected. The soluble extract of S. intermedius was separated by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were electrophoretically transferred onto nitrocellulose membrane. Western blotting for the specificity of serum IgG antistaphylococcal antibody was performed with anti-dog-IgG and sera obtained from an atopic dermatitis case and a normal dog. The molecular masses of four major proteins of S. intermedius recognized by serum obtained from an atopic dermatitis case were 18, 31, 75, and 110 kDa as determined by Western blot analysis. The present study indicates that most dogs of S. intermedius infection with atopic dermatitis could have a significant humoral immune response to bacterial proteins of the causative organism.

Panduratin A Inhibits Cell Proliferation by Inducing G0/G1 Phase Cell Cycle Arrest and Induces Apoptosis in Breast Cancer Cells

  • Liu, Qiuming;Cao, Yali;Zhou, Ping;Gui, Shimin;Wu, Xiaobo;Xia, Yong;Tu, Jianhong
    • Biomolecules & Therapeutics
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    • v.26 no.3
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    • pp.328-334
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    • 2018
  • Because of the unsatisfactory treatment options for breast cancer (BC), there is a need to develop novel therapeutic approaches for this malignancy. One such strategy is chemotherapy using non-toxic dietary substances and botanical products. Studies have shown that Panduratin A (PA) possesses many health benefits, including anti-inflammatory, anti-bacterial, anti-oxidant and anticancer activities. In the present study, we provide evidence that PA treatment of MCF-7 BC cells resulted in a time- and dose-dependent inhibition of cell growth with an $IC_{50}$ of $15{\mu}M$ and no to little effect on normal human MCF-10A breast cells. To define the mechanism of these anti-proliferative effects of PA, we determined its effect critical molecular events known to regulate the cell cycle and apoptotic machinery. Immunofluorescence and flow cytometric analysis of Annexin V-FITC staining provided evidence for the induction of apoptosis. PA treatment of BC cells resulted in increased activity/expression of mitochondrial cytochrome C, caspases 7, 8 and 9 with a significant increase in the Bax:Bcl-2 ratio, suggesting the involvement of a mitochondrial-dependent apoptotic pathway. Furthermore, cell cycle analysis using flow cytometry showed that PA treatment of cells resulted in G0/G1 arrest in a dose-dependent manner. Immunoblot analysis data revealed that, in MCF-7 cell lines, PA treatment resulted in the dose-dependent (i) induction of $p21^{WAF1/Cip1}$ and p27Kip1, (ii) downregulation of Cyclin dependent kinase (CDK) 4 and (iii) decrease in cyclin D1. These findings suggest that PA may be an effective therapeutic agent against BC.

Production of the polyclonal subunit C protein antibody against Aggregatibacter actinomycetemcomitans cytolethal distending toxin

  • Lee, Su-Jeong;Park, So-Young;Ko, Sun-Young;Ryu, So-Hyun;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.38 no.sup2
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    • pp.335-342
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    • 2008
  • Purpose: Cytolethal distending toxin (CDT) considered as a key factor of localized aggressive periodontitis, endocarditis, meningitis, and osteomyelitis is composed of five open reading frames (ORFs). Among of them, the individual role of CdtA and CdtC is not clear; several reports presents that CDT is an AB2 toxin and they enters the host cell via clathrin-coated pits or through the interaction with GM3 ganglioside. So, CdtA, CdtC, or both seem to be required for the delivery of the CdtB protein into the host cell. Moreover, recombinant CDT was suggested as good vaccine material and antibody against CDT can be used for neutralization or for a detection kit. Materials and Methods: We constructed the pET28a-cdtC plasmid from Aggregatibacter actinomycetemcomitans Y4 by genomic DNA PCR and expressed in BL21 (DE3) Escherichia coli system. We obtained the antibody against the recombinant CdtC in mice system. Using the anti-CdtC antibody, we test the native CdtC detection by ELISA and Western Blotting and confirm the expression time of native CdtC protein during the growth phase of A. actinomycetemcomitans. Results: In this study we reconstructed CdtC subunit of A. actinomycetemcomitans Y4 and generated the anti CdtC antibody against recombinant CdtC subunit expressed in E. coli system. Our anti CdtC antibody can be interacting with recombinant CdtC and native CDT in ELISA and Western system. Also, CDT holotoxin existed at 24h but not at 48h meaning that CDT holotoxin was assembled at specific time during the bacterial growth. Conclusion: In conclusion, we thought that our anti CdtC antibody could be used mucosal adjuvant or detection kit development, because it could interact with native CDT holotoxin.

Antibacterial Effects of Extracts from Citrus Peels (귤피 추출물의 항균효과)

  • Choi, Bo-Ram;Kang, Jaeku;Kang, Kyung-Hee
    • Journal of Digital Convergence
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    • v.10 no.11
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    • pp.559-564
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    • 2012
  • It has been reported that dried orange peel in a medicine prepared from crude drugs has antioxidant, anti-inflammatory, anti-allergic, anti-bacterial, and anti-viral functions, prevents circulatory diseases, reduces lipids, improves immunity, and strengthens capillary vessels. This study attempted to identify the antibacterial effects of dried orange peel extract on S. mutans causing dental caries, and its effects on GTase activation. The extract was put on the medium at the concentration of 5, 10 and 20 mg/ml and its effects on the inhibition of S. mutans growth were examined. Analysis found that the higher the concentration was, the more the number of colonies was reduced. In accordance with the measurement after 8 hours, the extract displayed a high growth inhibition rate: 92% at a concentration of 5 mg/ml, 95% at a concentration of 10 mg/ml and 99% at a concentration of 20 mg/ml. Furthermore, the GTase deactivation rate by the extract was measured. The higher the concentration of extract was, the more rapidly the deactivation rate increased. The deactivation rate of the extract was significant at 42% at a concentration of 5mg/ml.

Enhancement of the Antioxidant and Anti-inflammatory Activity of Hizikia fusiforme Water Extract by Lactic Acid Bacteria Fermentation (유산균 발효에 의한 톳 (Hizikia fusiforme) 추출액의 항산화 및 항염증 활성 증가)

  • Song, Ho-Su;Eom, Sung-Hwan;Kang, Young-Mi;Choi, Jong-Duck;Kim, Young-Mog
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.2
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    • pp.111-117
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    • 2011
  • We previously reported that fermentation by Lactobacillus brevis LB-20 isolated from Kimchi resulted in improvement of the sensory quality of Hizikia fusiforme water extract. This study was conducted to evaluate the possible application of lactic acid bacteria fermentation to improve the functional qualities of H. fusiforme extract. L. brevis LB-20 was inoculated and cultivated in H. fusiforme extract. The antioxidant and anti-inflammatory activities of extract were then assayed both before and following fermentation for two days. Antioxidant activity was determined by assaying levels of radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, superoxide, and alkyl radical. Lactic acid bacterial fermentation of H. fusiforme extract resulted in enhancement of antioxidant activity. The greatest enhancement of antioxidant activity was seen in the hydroxyl radical scavenging assay that incorporated 0.5 mg/mL of raw and fermented H. fusiforme extract. Fermented extract exhibited greater (21.95%) inhibition of nitric oxide synthesis than did raw extract (14.66%) at a concentration of 1 mg/mL. The fermented extract exerted its potent anti-inflammatory activity via attenuation of expression of inflammation-related cytokine proteins (TNF-${\alpha}$ and iNOS).

A Comparison of Two Methods for the Extraction of Lactoferrin-binding Proteins from Streptococcus uberis (Streptococcus uberis의 락토페린 결합단백질 추출을 위한 두 가지 방법의 비교)

  • Park, Hee-Myung;Yoo, Jong-Hyun;Almeida, Raul A.;Oliver, Stephen P.
    • Journal of Veterinary Clinics
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    • v.24 no.3
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    • pp.305-307
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    • 2007
  • Lactoferrin-binding proteins (LBP) has not been well characterized in Streptococcus uberis isolated from milk of bovine mastitis and to date this protein is considered to be an important virulence factor in Streptococcal mastitis. To determine the more efficient extraction method of LBP from four S. uberis strains, we used two different extraction methods (mutanolysin and sodium dodecyl sulfate) in this study. Bacterial proteins extracted were electrophoresed by 10% polyacrylamide gels in the presence of sodium deodecyl sulfate and gels were transferred onto nitrocellulose membrane. Rabbit anti-bovine lactoferrin antibody and HRP-conjugated donkey anti-rabbit IgG antibody were used to detect LBP. This Western blotting analysis demonstrates that extraction method with SDS extracted 110 kDa and 112 kDa LBPs more efficiently compared to the mutanolysin extraction method.