Antibacterial mechanism of high-mobility group nucleosomal-binding domain 2 on the Gram-negative bacteria Escherichia coli.

OBJECTIVE: To investigate the antibacterial mechanism of high-mobility group nucleosomal-binding domain 2 (HMGN2) on Escherichia coli K12, focusing on the antibacterial and antibiofilm formation effects. Its chemotactic activity on human neutrophils was also investigated. METHODS: Human tissue-derived HMGN2 (tHMGN2) was extracted from fresh uterus fiber cystadenoma and purified by HP1100 reversed-phase high-performance liquid chromatography (RP-HPLC). Recombinant human HMGN2 (rHMGN2) was generated in E. coli DE3 carrying PET-32a-c(+)-HMGN2. Antibacterial activity of HMGN2 was determined using an agarose diffusion assay and minimum inhibitory concentration (MIC) of HMGN2 was determined by the microdilution broth method. Bacterial membrane permeability assay and DNA binding assay were performed. The antibiofilm effect of HMGN2 was investigated using a crystal violet assay and electron microscopy scanning. The activating effect and chemotactic activity of HMGN2 on neutrophils were determined using a nitroblue tetrazolium (NBT) reduction assay and Transwell chamber cell migration assay, respectively. RESULTS: HMGN2 showed a relatively high potency against Gram-negative bacteria E. coli and the MIC of HMGN2 was 16.25 µg/ml. Elevated bacterial membrane permeability was observed in HMGN2-treated E. coli K12. HMGN2 could also bind the bacterial plasmid and genomic DNA in a dose-dependent manner. The antibiofilm effect of HMGN2 on E. coli K12 was confirmed by crystal violet staining and scanning electron microscopy. However, the activating effects and chemotactic effects of HMGN2 on human neutrophils were not observed. CONCLUSIONS: As an antimicrobial peptide (AMP), HMGN2 possessed a good capacity for antibacterial and antibiofilm activities on E. coli K12. This capacity might be associated with disruption of the bacterial membrane and combination of DNA, which might affect the growth and viability of E. coli.

Multiple scattering of electromagnetic waves by an aggregate of uniaxial anisotropic spheres.

An exact analytical solution is obtained for the scattering of electromagnetic waves from a plane wave with arbitrary directions of propagation and polarization by an aggregate of interacting homogeneous uniaxial anisotropic spheres with parallel primary optical axes. The expansion coefficients of a plane wave with arbitrary directions of propagation and polarization, for both TM and TE modes, are derived in terms of spherical vector wave functions. The effects of the incident angle α and the polarization angle ß on the radar cross sections (RCSs) of several types of collective uniaxial anisotropic spheres are numerically analyzed in detail. The characteristics of the forward and backward RCSs in relation to the incident wavelength are also numerically studied. Selected results on the forward and backward RCSs of several types of square arrays of SiO2 spheres illuminated by a plane wave with different incident angles are described. The accuracy of the expansion coefficients of the incident fields is verified by comparing them with the results obtained from references when the plane wave is degenerated to a z-propagating and x- or y-polarized plane wave. The validity of the theory is also confirmed by comparing the numerical results with those provided by a CST simulation.

[Study on antimicrobial activity of human hemoglobin and its fragments both in vitro and in vivo].

OBJECTIVE: To isolate human hemoglobin and its fragrments, compare their antimicrobial activity in vitro and pilot study of their antimicrobial activity in vivo. METHODS: The alpha and beta chains of hemoglobin were separated by cation exchange chromatography and gel chromatography; The alpha and beta chains were cleaved by cyanogens bromide respectively. The cleaved fragments were purified by reverse phase high performance liquid chromatography. Antimicrobial activity of hemoglobin and its fragments was determined by agrose radial diffusion assay. After establishment of E. coli vaginal infection model, the rats were randomized into the experimental group (hemoglobin group) and the control group. The histologically pathological section was observed. RESULTS: Hemoglobin, alpha/beta chain and their fragments had similar antibacterial activities in vitro, which were mainly against Gram-negative bacteria E. coli; except alpha1-32 had a comparatively lower activity. Antimicrobial activity in vivo: a comparison of the hemoglobin group with the matrix control group (no treatment after infection), the surface layer of vaginal stratified squamous epithelium was smoother, inflammatory cells were significantly reduced in the lamina propria and congestion was obviously decreased. CONCLUSION: Human hemoglobin and its fragments had antibacterial activity in vitro, hemoglobin might relieve the inflammation of E. coil vaginal infection in rats moreover.