Journal of Microbiology

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Functional Characterization of DNA N‑Glycosylase Ogg1 and Ntg1 in DNA Damage Stress of Cryptococcus neoformans: Kwang-Woo Jung , Sunhak Kwon , Jong-Hyun Jung , Sangyong Lim , Yong-Sun Bahn; J. Microbiol. 2023;61(11):981-992. Published online December 6, 2023; DOI: https://doi.org/10.1007/s12275-023-00092-y

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Abstract: Reactive oxygen species induce DNA strand breaks and DNA oxidation. DNA oxidation leads to DNA mismatches, resulting in mutations in the genome if not properly repaired. Homologous recombination (HR) and non-homologous end-joining (NHEJ) are required for DNA strand breaks, whereas the base excision repair system mainly repairs oxidized DNAs, such as 8-oxoguanine and thymine glycol, by cleaving the glycosidic bond, inserting correct nucleotides, and sealing the gap. Our previous studies revealed that the Rad53-Bdr1 pathway mainly controls DNA strand breaks through the regulation of HRand NHEJ-related genes. However, the functional roles of genes involved in the base excision repair system remain elusive in Cryptococcus neoformans. In the present study, we identified OGG1 and NTG1 genes in the base excision repair system of C. neoformans, which are involved in DNA oxidation repair. The expression of OGG1 was induced in a Hog1-dependent manner under oxidative stress. On the other hand, the expression of NTG1 was strongly induced by DNA damage stress in a Rad53-independent manner. We demonstrated that the deletion of NTG1, but not OGG1, resulted in elevated susceptibility to DNA damage agents and oxidative stress inducers. Notably, the ntg1Δ mutant showed growth defects upon antifungal drug treatment. Although deletion of OGG1 or NTG1 did not increase mutation rates, the mutation profile of each ogg1Δ and ntg1Δ mutant was different from that of the wild-type strain. Taken together, we found that DNA N-glycosylase Ntg1 is required for oxidative DNA damage stress and antifungal drug resistance in C. neoformans.

Lipoteichoic acids of lactobacilli inhibit Enterococcus faecalis biofilm formation and disrupt the preformed biofilm: Solmin Jung , Ok-Jin Park , A Reum Kim , Ki Bum Ahn , Dongwook Lee , Kee-Yeon Kum , Cheol-Heui Yun , Seung Hyun Han; J. Microbiol. 2019;57(4):310-315. Published online January 22, 2019; DOI: https://doi.org/10.1007/s12275-019-8538-4

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Enterococcus faecalis, a Gram-positive bacterium commonly isolated in patients with refractory apical periodontitis, invades dentin tubules easily and forms biofilms. Bacteria in biofilms, which contribute to recurrent and/or chronic inflammatory diseases, are more resistant to antimicrobial agents than planktonic cells and easily avoid phagocytosis. Although Lactobacillus plantarum lipoteichoic acid (Lp.LTA) is associated with biofilm formation, the effect of Lp.LTA on biofilm formation by E. faecalis is not clearly understood. In this study, we investigated whether Lp.LTA inhibits E. faecalis biofilm formation. The degree of biofilm formation was determined by using crystal violet assay and LIVE/DEAD bacteria staining. The quantification of bacterial growth was determined by measuring the optical density at 600 nm with a spectrophotometer. Formation of biofilms on human dentin slices was observed under a scanning electron microscope. E. faecalis biofilm formation was reduced by Lp.LTA treatment in a dose-dependent manner. Lp.LTA inhibited biofilm development of E. faecalis at the early stage without affecting bacterial growth. LTA from other Lactobacillus species such as Lactobacillus acidophilus, Lactobacillus casei, or Lactobacillus rhamnosus GG also inhibited E. faecalis biofilm formation. In particular, among LTAs from various lactobacilli, Lp.LTA showed the highest inhibitory effect on biofilms formed by E. faecalis. Interestingly, LTAs from lactobacilli could remove the biofilm preformed by E. faecalis. These inhibitory effects were also observed on the surface of human dentin slices. In conclusion, Lactobacillus species LTA inhibits biofilm formation caused by E. faecalis and it could be used as an anti-biofilm agent for prevention or treatment against E. faecalis-associated diseases.

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Macrolide Resistance and In Vitro Selection of Resistance to Antibiotics in Lactobacillus Isolates: Lorenzo Drago , Roberto Mattina , Lucia Nicola , Valentina Rodighiero , Elena De Vecchi; J. Microbiol. 2011;49(4):651-656. Published online September 2, 2011; DOI: https://doi.org/10.1007/s12275-011-0470-1

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Abstract: Spreading of resistance to antibiotics is of great concern due to the increasing rate of isolation of multiresistant pathogens. Since commensal bacteria may transfer determinants of resistance to pathogens, studies on development of resistance should include also lactobacilli. Resistance to macrolides, penicillins and tetracycline was determined in 40 isolates of Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus crispatus, and Lactobacillus casei isolated from faeces of apparently healthy volunteers. Frequency of mutation and changes in susceptibility after serial exposure to these antibiotics at concentrations of 4× and 8× MIC were evaluated in susceptible isolates. Acquired resistance was defined as an increment in MIC values of at least four times in respect to the pre-selection values. Resistance to macrolides and/or tetracycline was identified in 14 and 4 isolates, respectively. ermB gene and A2058G mutation in 23S rRNA were detected in macrolide resistant isolates. Frequencies of mutation of susceptible isolates (n=26) were lower for ampicillin and erythromycin than for tetracycline. Serial exposure to antibiotics led to selection of resistant mutants. However, acquired resistance was rather unstable and was lost after subcultures in antibiotic-free medium in most mutants. Resistance to erythromycin was associated to a A2058G mutation in 23S rRNA. In conclusion,
results
indicate that resistance to macrolides and tetracycline is present among intestinal lactobacilli. Decrease in susceptibility following serial exposure to antibiotics might occur in lactobacilli, in a strain- and antibiotic- dependent way. Since lactobacilli are often used as probiotics, their ability to acquire resistance should be evaluated for isolates candidate to be included in probiotics based products.

Induction of Cytokines and Nitric Oxide in Murine Macrophages Stimulated with Enzymatically Digested Lactobacillus Strains: Dong Woon Kim , Sung Back Cho , Cheol Heui Yun , Ha Yeon Jeong , Wan Tae Chung , Chang Weon Choi , Hyun Jeong Lee , In Sik Nam , Guk Hyun Suh , Sang Suk Lee , Byong Seak Lee; J. Microbiol. 2007;45(5):373-378.; DOI: https://doi.org/2601 [pii]

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Abstract: Based on observations that lactic acid bacteria have the ability to activate macrophages, we assessed the potential effects of eight different Lactobacillus strains treated with gastrointestinal enzymes on the production of nitric oxide and various cytokines in macrophages. RAW 264.7 murine macrophage cells were cultured with either precipitates or supernatants of Lactobacillus strains digested with pepsin followed by pancreatin. The increased production of nitric oxide and interleukin (IL)-1β, IL-6, IL-12 and tumour necrosis factor (TNF)-α were observed when cultured with precipitates, and this effect was largely strain-dependent. In contrast, the exposure of RAW 264.7 cells to supernatants produced weaker or nearly undetectable effects in comparison to the effects of exposure to precipitates. The induction of nitric oxide appeared to be unaffected. These results demonstrate that nitric oxide and cytokines were effectively induced when the bacterial precipitate was treated with macrophages. The results of the present study also indicate that Lactobacillus strains treated with digestive enzymes are capable of stimulating the production of nitric oxide and cytokines in macrophages, which may modulate the gastrointestinal immune function of the host when it is given as a feed additive.

Antibacterial Activities of Lactobacillus crispatus ATCC 33820 and Lactobacillus gasseri ATCC 33323: Jin-Woo Kim , S.N. Rajagopal; J. Microbiol. 2001;39(2):146-148.

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Abstract: Lactobacillus crispatus ATCC 33820 and L. gasseri ATCC 33323 were grown in MRS broth (pH 6.5) at 37 C for 24 h and the antibacterial activities of cell free culture supernatants were determined by the agar well diffusion method. The culture supernatants were inhibitory to Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, Enterococcus faecalis, Pediococcus acidilacticii, and Lactobacillus helveticus. The supernatants did not show any lysozyme activity. Addition of catalase did not affect the antibacterial activities of the supernatants. The antibacterial substances were heat stable (100 C for 60 min) and sensitive to proteases.

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