Covalent Complex of DNA and Bacterial Topoisomerase: Implications in Antibacterial Drug Development

A topoisomerase-DNA transient covalent complex can be a druggable target for novel topoisomerase poison inhibitors that represent a new class of antibacterial or anticancer drugs. Herein, we have investigated molecular features of the functionally important Escherichia coli topoisomerase I (EctopoI)-DNA covalent complex (EctopoIcc) for molecular simulations, which is very useful in the development of new antibacterial drugs. To demonstrate the usefulness of our approach, we used a model small molecule (SM), NSC76027, obtained from virtual screening. We examined the direct binding of NSC76027 to EctopoI as well as inhibition of EctopoI relaxation activity of this SM via experimental techniques. We then performed molecular dynamics (MD) simulations to investigate the dynamics and stability of EctopoIcc and EctopoI-NSC76027-DNA ternary complex. Our simulation results show that NSC76027 forms a stable ternary complex with EctopoIcc. EctopoI investigated here also serves as a model system for investigating a complex of topoisomerase and DNA in which DNA is covalently attached to the protein.     

Antibacterial mechanism of lactic acid on physiological and morphological properties of Salmonella Enteritidis,Escherichia coli and Listeria monocytogenes

Lactic acid is widely used to inhibit the growth of important microbial pathogens, but its antibacterial mechanism is not yet fully understood. The objective of this study was to investigate the antibacterial mechanism of lactic acid on Salmonella Enteritidis, Escherichia coli and Listeria monocytogenes by size measurement, TEM, and SDS-PAGE analysis. The results indicated that 0.5% lactic acid could completely inhibit the growth of Salmonella Enteritidis, E. coli and L. monocytogenes cells. Meanwhile, lactic acid resulted in leakage of proteins of Salmonella, E. coli and Listeria cells, and the amount of leakage after 6 h exposure were up to 11.36, 11.76 and 16.29 μg/mL, respectively. Measurements of the release of proteins and SDS-PAGE confirmed the disruptive action of lactic acid on cytoplasmic membrane, as well as the content and activity of bacterial proteins. The Z-Average sizes of three pathogens were changed to smaller after lactic acid treatment. The damaged membrane structure and intracellular structure induced by lactic acid could be observed from TEM images. The results suggested that the antimicrobial effect was probably caused by physiological and morphological changes in bacterial cells.     

Coconut husk extract: antibacterial properties and its application for shelf-life extension of Asian sea bass slices

Antibacterial properties of ethanolic coconut husk extracts (ECHE) prepared using varying ethanol concentrations as the extracting media against Pseudomonas aeruginosa, Escherichia coli, Vibrio parahaemolyticus, Listeria monocytogenes and Staphylococcus aureus was studied. ECHE60, prepared using 60% ethanol, had similar minimum inhibitory concentration and minimum bactericidal concentration to those extracted using 80% and 100% ethanol (P ˂ 0.05). When Asian sea bass (Lates calcarifer) slices were treated with ECHE60 at 200 and 400 ppm, the quality changes were monitored in comparison with the control (without treatment) during 12 days of refrigerated storage (4 °C). The shelf life of sea bass treated with 400 ppm ECHE60 was extended to 9 days, whereas the control had a shelf life of 3 days. Lipid oxidation was lowered in ECHE60 treated samples than the control during storage. Therefore, ECHE60 could inhibit both spoilage and pathogenic bacteria and also extended the shelf life of sea bass slices.     

Precise quantification of the antibacterial activity of chitosan by NB medium neutralizer

In the present research,nutrient broth(NB)medium was identified to be able to neutralize the antibac-terial activity of chitosan and its derivatives.Therefore,an improved test method independent of NB medium was proposed to precisely quantify the antibacterial effectiveness and efficiency of chitosan.The minimum bactericidal concentration(MBC)of chitosan was 60 μg mL-1 against S.aureus and E.coli,and 0.01％(w/v)chitosan could kill 100％of bacteria within 3 min.From another point of view,the neutralizing efficiency of NB could be tripled by adding 25 g L-1 of sodium chloride.Then the neutraliz-ing mechanism of NB medium was ascribed to flocculation between chitosan and protein.Adding extra sodium chloride could significantly reduce the size of floccules,and smaller floccules would lose the ability of binding with bacteria directly,showing higher neutralizing rate on the macro scale.     

Metallocenyl 7-ACA Conjugates: Antibacterial Activity Studies and Atomic-Resolution X-ray Crystal Structure with CTX-M β-Lactamase

The conjugation of organometallic groups to current β-lactam antibiotics is a field of increasing study due to the ability of certain organometallic groups to enhance the antibiotic potency of these drugs. Herein, we report the antibacterial properties of two metallocenyl (ferrocenyl and ruthenocenyl) 7-aminocephalosporanic acid (7-ACA) antibiotic conjugates. Continuing a trend we found in our previous studies, the ruthenocenyl conjugate showed greater antibacterial activity than its ferrocenyl counterpart. Compared with the previously published 7-aminodesacetoxycephalosporanic acid (7-ADCA) conjugates, the 3-acetyloxymethyl group significantly improved the compounds’ activity. Furthermore, the Rc-7-ACA compound was more active against clinical Staphylococcus aureus isolates than the ampicillin reference. Noticeably, neither of the two new compounds showed an undesirable toxic effect in HeLa and L929 cells at the concentrations at which they displayed strong antibacterial effects. The antibacterial activity of the two metallocenyl 7-ACA derivatives was further confirmed by scanning electron microscopy (SEM). SEM micrographs showed that bacteria treated with metallocenyl 7-ACA derivatives feature cell wall damage and morphology changes. Using a CTX-M-14 β-lactamase competition assay based on nitrocefin hydrolysis, we showed that the Rc-7-ACA bound more favorably to CTX-M-14 than its ferrocenyl counterpart, again confirming the superiority of the ruthenocenyl moiety over the ferrocenyl one in interacting with proteins. We also report a 1.47 Å resolution crystal structure of Rc-7-ACA in complex with the CTX-M-14 E166A mutant, an enzyme sharing a similar active site configuration with penicillin-binding proteins, the molecular target of β-lactam antibiotics. These results strengthen the case for the antibacterial utility of the Rc and Fc groups.     

Facile fabrication of hollow mesoporous bioactive glass spheres: From structural behaviour to in vitro biology evaluation

Bone defects accompanied by infection or inflammation can significantly delay the healing process. To simultaneously achieve controlled release of local antibiotics for infection control and bone healing, bone-implantable delivery systems have been considered as a promising strategy. This study aims to improve drug loading capacity of bone-implantable delivery systems by introducing hollow structure mesoporous bioactive glass nanospheres (HMBGs) through a sol–gel process. Particularly, such core–shell bimodal-porous structured nanoparticles were prepared through a sacrificing template using cetyltrimethylammonium bromide (CTAB) as surfactant. It was found that varying the amount of CTAB during the synthesis process is a simple and effective approach for tuning the particle size, morphology, and structure of HMBGs. For in vitro drug release, HMBGs could sustain storage and release of vancomycin hydrochloride (VAN) via diffusion-controlled mechanism, thereby inhibiting the bacteria growth in the subsequent bacterial study. Moreover, HMBGs incorporated with VAN provided a biomimetic microenvironment favored by cell adhesion and proliferation. These findings support the compatibility of HMBG nanoparticles with antibiotics and their potential application in the treatment of infectious bone defects.     

Development of compartmentalized antibacterial systems based on encapsulated alliinase

Allicin, an organic compound produced via the transformation of biologically inactive alliin by alliinase, an enzyme found in garlic, combines a strong antibacterial effect with suppressed development of bacterial resistance. However, because of the high reactivity and volatility, controlled in-situ production of allicin that mimics the natural synthesis is essential to achieve desired therapeutic effects. In this work, the spray-drying technique was employed for encapsulation of alliinase into polymer micro-carriers with an emphasis on the effect of process parameters, i.e., drying temperature, nozzle type, choice of the carrier materials, on the activity of encapsulated alliinase in soluble maltodextrin and swellable chitosan microparticles. The results show that maltodextrin is a suitable carrier for the effective protection of alliinase against thermal stress. On the other hand, we can control allicin production and release from chitosan carriers with immobilized alliinase by variation of the cross-linker amount. Antibacterial activity of in-situ formed allicin vapors was confirmed against Escherichia coli bacterial strain using customized sample holders preventing physical contact of powder sample with the inoculated agar plate.     

Evaluation of antibacterial protein with antioxidant activity from Rahnella aquatilis L103 and its effect on beef during refrigerated storage

A strain, Rahnella aquatilis L103, which was isolated from the soil around the roots of mushrooms, produced antibacterial protein though fermentation. This protein has broad antibacterial spectrum towards gram-positive and gram-negative bacteria. The protein has a significant antioxidant capacity on scavenging ABTS^+· (82.5%, 120 μg mL⁻¹), DPPH^· (64.1%, 600 μg mL⁻¹) and OH^· (60.1%, 750 μg mL⁻¹) as well. The protein was considered to be safe within concentration of 150 μg mL⁻¹ as shown by MTT results and was applied to beef refrigerated preservation. The treatment group presented lower total viable counts (TVC), total volatile basic nitrogen (TVB-N), 2-thiobarbituric acid (TBA), meanwhile, higher sensory score and longer shelf life (2 days) compared with the control group (P < 0.05). The results indicated that the protein under safe concentration was beneficial to beef preservation.     

Hydroxyapatite forming ability,ion release and antibacterial activity of the melt-derived SiO2–P2O5–Na2O–CaO–F glasses modified by replacing CaO with SrO and ZnO

Since the discovery of 1970s, bioactive glass has been a hot topic of research because of its excellent biological activity, which makes it a material that can repair and replace human bone tissue organs. In this work, the bioactive glasses in the system SiO₂–P₂O₅–Na₂O–CaO–F with different amounts of strontium oxide (SrO) and zinc oxide (ZnO) were prepared by the conventional melt quenching technology. The hydroxyapatite (HA) forming ability, ion release and antibacterial activity of these prepared glasses were investigated and the obtained results illustrated that SrO-doped samples had a better ability to form HA in modified simulated body fluid (MSBF) than ZnO-doped samples. As the immersion time of the sample in MSBF increased, the content of HA phase gradually increased. In the same immersion time, the formation ability of HA and the variation of SrO substitution amount showed a non-linear trend, which is mainly related to the influence of SrO content on the glass network structure. The results of ion concentration showed that the formation of HA was the result of the comprehensive action of various ions in the solution, especially the release rate of Si⁴⁺ ions, which had a direct impact on the formation ability of HA. The antibacterial test illustrated that the difference in antibacterial activity of bacteria solution at different sample concentrations may be related to the high pH environment and the osmotic effects caused by the non-physiological concentration of ions in the solution. The glass sample contained 4 wt% SrO showed the minimum bactericidal concentration at 64 mg/mL. The glass samples prepared in this experiment had good biological activity and antibacterial effect, making them suitable for using in dentistry and orthopedic applications, as well as providing a valuable composition reference for the preparation of bioactive glass with excellent comprehensive properties.     

A high strength,wear and corrosion-resistant,antibacterial and biocompatible Nb-5 at.％ Ag alloy for dental and orthopedic implants

Refractory metal niobium (Nb) incorporated with a small amount of silver (Ag),the resulting Nb-Ag two-phase alloys,were fabricated by mechanical alloying and spark plasma sintering.The microstructure,mechanical properties,wear resistance,corrosion behavior,in vitro and in vivo antibacterial properties and biocompatibility of the Nb-Ag alloys were systematically investigated.The results show that the mechanical properties,wear resistance,corrosion resistance and antibacterial ability were significantly enhanced after addition of 5 at.％ Ag.The fabricated Nb-5 at.％ Ag alloy demonstrates high yield strength of up to ～ 1486 MPa and fracture strain of ～ 35 ％.The precipitated Ag particles could reduce friction and wear.The enhanced corrosion resistance was attributed to the higher relative density of the sintered alloys and the formation of a stable and dense passive film of niobium and silver oxides.In vitro and in vivo evaluations show that the Nb-5 at.％ Ag alloy also has strong antibacterial activity and good biocompati-bility and osteointegration ability.These results demonstrate great potential of the nanostructured Nb-Ag alloys for dental and orthopedic implants.