The use of lysozyme modified with fluorescein for the detection of Gram-positive bacteria

Arabski, Michał; Konieczna, Iwona; Tusińska, Ewa; Wa̡sik, Sławomir; Relich, Inga; Zając, Krzysztof; Kamiński, Zbigniew J.; Kaca, Wiesław

doi:10.1016/j.micres.2014.05.004

Cited by 23 publications

(10 citation statements)

References 22 publications

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“…Lysozyme is the major component of the innate immune system against infection by bacterial pathogens, particularly Gram-positive bacteria [18]. We analyzed the effect of baicalin on LYSO expression using qPCR and western blotting (Figure 2).…”

Section: Resultsmentioning

confidence: 99%

Baicalin promotes the bacteriostatic activity of lysozyme on S. aureus in mammary glands and neutrophilic granulocytes in mice

et al. 2017

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Staphylococcus aureus causes mastitis as a result of community-acquired or nosocomial infections. Lysozyme (LYSO) is an enzyme that is upregulated in many organisms during the innate immune response against infection by bacterial pathogens. Baicalin is a bioactive flavonoid that can bind to enzymes, often to potentiate their effect. Here we tested the effects of baicalin on the activity of LYSO using the S. aureus mastitis mouse model and neutrophilic granulocyte model of S. aureus infection. In our experiments, S. aureus counts decreased with increasing baicalin concentration. Furthermore, qPCR and western blot analyses showed that LYSO expression was unaffected by baicalin, while fluorescence quenching and UV fluorescence spectral analyses showed that baicalin binds to LYSO. To test whether this binding increased LYSO activity, we assessed LYSO-induced bacteriostasis in the presence of baicalin. Our results showed that LYSO-induced S. aureus bacteriostasis increased with increasing concentrations of baicalin, and that baicalin binding to LYSO synergistically increased the antibacterial activity of LYSO. These results demonstrate that baicalin enhances LYSO-induced bacteriostasis during the innate immune response to S. aureus. They suggest baicalin is a potentially useful therapeutic agent for the treatment of bacterial infections.

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Section: Resultsmentioning

confidence: 99%

Baicalin promotes the bacteriostatic activity of lysozyme on S. aureus in mammary glands and neutrophilic granulocytes in mice

et al. 2017

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“…To date, the most widely used diagnostics is the plate-count method, based on the growth of bacteria on an agar plate in the presence of different antibiotics [3]. This technique is timeconsuming (24-72 hours) because it requires many cell-division cycles; expert users are also required for sample preparation and final analysis [4]. The time-delay is the critical issue, however, because some infections such as sepsis require immediate attention [5] and, more generally, clinicians administer antibiotics based on experience rather than accurate diagnosis, which has a significant error rate.…”

Section: Introductionmentioning

confidence: 99%

Monitoring of individual bacteria using electro-photonic traps

Conteduca¹,

Brunetti²,

Dell’Olio³

et al. 2019

Biomed. Opt. Express

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Antimicrobial resistance (AMR) describes the ability of bacteria to become immune to antimicrobial treatments. Current testing for AMR is based on culturing methods that are very slow because they assess the average response of billions of bacteria. In principle, if tests were available that could assess the response of individual bacteria, they could be much faster. Here, we propose an electro-photonic approach for the analysis and the monitoring of susceptibility at the single-bacterium level. Our method employs optical tweezers based on photonic crystal cavities for the trapping of individual bacteria. While the bacteria are trapped, antibiotics can be added to the medium and the corresponding changes in the optical properties and motility of the bacteria be monitored via changes of the resonance wavelength and transmission. Furthermore, the proposed assay is able to monitor the impedance of the medium surrounding the bacterium, which allows us to record changes in metabolic rate in response to the antibiotic challenge. For example, our simulations predict a variation in measurable electrical current of up to 40% between dead and live bacteria. The proposed platform is the first, to our knowledge, that allows the parallel study of both the optical and the electrical response of individual bacteria to antibiotic challenge. Our platform opens up new lines of enquiry for monitoring the response of bacteria and it could lead the way towards the dissemination of a new generation of antibiogram study, which is relevant for the development of a point-of-care AMR diagnostics.

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“…3.9 nm which plays a crucial role in targeting Gram positive bacteria as a first line of defense in the human body. 43,44 Due to this, lysozyme is found abundantly in the nasal and ocular membranes and its deficiency often results in conjunctivitis or other increased incidences in illness. 45 The applicability of the temperature controlled delivery of lysozyme from CS-MSNs is tested by observing its antibacterial properties on Gram positive Bacillus cereus and Micrococcus luteus bacteria.…”

Section: Introductionmentioning

confidence: 99%

Poly(N-isopropylacrylamide)-gated Fe3O4/SiO2 core shell nanoparticles with expanded mesoporous structures for the temperature triggered release of lysozyme

Galiana

Martínez‐Máñez

et al. 2015

Colloids and Surfaces B: Biointerfaces

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Core-shell nanoparticles comprised of Fe3O4 cores and a mesoporous silica shell with an average expanded pore size of 6.07 nm and coated with a poly(N-isopropylacrylamide) (PNIPAM) layer (CS-MSNs-EP-PNIPAM) were prepared and characterized. The nanoparticles was loaded with (Ru(bipy)3(2+)) dye or an antibacterial enzyme, lysozyme, to obtain CS-MSNs-EP-PNIPAM-Ru(bipy)3(2+) and CS-MSNs-EP-PNIPAM-Lys, respectively. The lysozyme loading was determined to be 160 mg/g of nanoparticle. It was seen that Ru(bipy)3(2+) and lysozyme release was minimal at a room temperature of 25 °C while at physiological temperature (37 °C), abrupt release was observed. The applicability of the CS-MSNs-EP-PNIPAM-Lys was further tested with two Gram-positive bacteria samples, Bacillus cereus and Micrococcus luteus. At physiological temperature, the nanoparticles were shown to reduce bacterial growth, indicating a successful release of lysozyme from the nanoparticles. This nanoparticle system shows potential as a nanocarrier for the loading of similarly sized proteins or other species as a drug delivery platform.

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The use of lysozyme modified with fluorescein for the detection of Gram-positive bacteria

Cited by 23 publications

References 22 publications

Baicalin promotes the bacteriostatic activity of lysozyme on S. aureus in mammary glands and neutrophilic granulocytes in mice

Baicalin promotes the bacteriostatic activity of lysozyme on S. aureus in mammary glands and neutrophilic granulocytes in mice

Monitoring of individual bacteria using electro-photonic traps

Poly(N-isopropylacrylamide)-gated Fe3O4/SiO2 core shell nanoparticles with expanded mesoporous structures for the temperature triggered release of lysozyme

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