Neutralizing Staphylococcus aureus Virulence with AZD6389, a Three mAb Combination, Accelerates Closure of a Diabetic Polymicrobial Wound.

Nonhealing diabetic foot ulcers (DFU), a major complication of diabetes, are associated with high morbidity and mortality despite current standard of care. Since Staphylococcus aureus is the most common pathogen isolated from nonhealing and infected DFU, we hypothesized that S. aureus virulence factors would damage tissue, promote immune evasion and alter the microbiome, leading to bacterial persistence and delayed wound healing. In a diabetic mouse polymicrobial wound model with S. aureus, Pseudomonas aeruginosa, and Streptococcus pyogenes, we report a rapid bacterial proliferation, prolonged pro-inflammatory response and large necrotic lesions unclosed for up to 40 days. Treatment with AZD6389, a three-monoclonal antibody combination targeting S. aureus alpha toxin, 4 secreted leukotoxins, and fibrinogen binding cell-surface adhesin clumping factor A resulted in full skin re-epithelization 21 days after inoculation. By neutralizing multiple virulence factors, AZD6389 effectively blocked bacterial agglutination and S. aureus-mediated cell killing, abrogated S. aureus-mediated immune evasion and targeted the bacteria for opsonophagocytic killing. Neutralizing S. aureus virulence not only facilitated S. aureus clearance in lesions, but also reduced S. pyogenes and P. aeruginosa numbers, damaging inflammatory mediators and markers for neutrophil extracellular trap formation 14 days post initiation. Collectively, our data suggest that AZD6389 holds promise as an immunotherapeutic approach against DFU complications. IMPORTANCE Diabetic foot ulcers (DFU) represent a major complication of diabetes and are associated with poor quality of life and increased morbidity and mortality despite standard of care. They have a complex pathogenesis starting with superficial skin lesions, which often progress to deeper tissue structures up to the bone and ultimately require limb amputation. The skin microbiome of diabetic patients has emerged as having an impact on DFU occurrence and chronicity. DFU are mostly polymicrobial, and the Gram-positive bacterium Staphylococcus aureus detected in more than 95% of cases. S. aureus possess a collection of virulence factors which participate in disease progression and may facilitate growth of other pathogens. Here we show in a diabetic mouse wound model that targeting some specific S. aureus virulence factors with a multimechanistic antibody combination accelerated wound closure and promoted full skin re-epithelization. This work opens promising new avenues for the treatment of DFU.

[Preparation of Modified Watermelon Biochar and Its Adsorption Properties for Pb(â ¡)].

In this study, watermelon rind was used as a raw material to modify watermelon rind biochar (MBC) with ammonium sulphate[(NH4)2S] for adsorption of Pb(Ⅱ) ions. The effects of solution pH, adsorption time, adsorbent addition amount, initial mass concentration of Pb(Ⅱ) ions, and ionic strength on the adsorption of Pb(Ⅱ) ions were investigated. The results show that the saturated adsorption time was 5 h, the optimum pH of the adsorption reaction was 6, and when the initial mass concentration of Pb(Ⅱ) ions were 1000 mg·L-1, and the amount of adsorbent was 2.0 g·L-1. The maximum adsorption amount of MBC to Pb(Ⅱ) ions can reach 97.63 mg·g-1, which is significantly higher than unmodified watermelon husk biochar (BC). The adsorption of Pb(Ⅱ) ions by modified watermelon biochar was in accordance with the Langmuir isotherm adsorption model and the pseudo second-order kinetic model, which proves that adsorption is dominated by monolayer chemical adsorption. The desorption of MBC after adsorption of Pb(Ⅱ) ions was carried out using a sodium hydroxide solution to study the reusability of MBC, and the adsorption amount was still 64.74 mg·g-1 in the sixth cycle. Characterization and analysis of adsorbents by Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, nitrogen adsorption, scanning electron microscopy-energy spectroscopy, zeta potential analysis, and X-ray diffraction (XRD) were carried out, which showed that the adsorption mechanism is mainly that MBC oxygen- and MBC sulfur-containing groups adsorb Pb(Ⅱ) through complexation and precipitation. Therefore, ammonium sulfide modified watermelon rind biochar can be used as a highly efficient lead adsorbent.

[Effects and Mechanisms of Methyl Orange Removal from Aqueous Solutions by Modified Rice Shell Biochar].

Waste rice shell (RS) was used for modified biochar preparation via different activation methods. The types of modifiers, impregnation ratio, and pyrolysis temperature have significant effects on the characteristics of biochar and the adsorption capacity of methyl orange (MO). The physical and chemical properties of modified biochar and MO adsorption mechanisms were analyzed by N2-adsorption, X-ray diffraction (XRD), Fourier infrared spectroscopy (FT-IR), field emission scanning electron microscopy (SEM), thermogravimetric analyzer (TG), transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS) techniques. The results showed that the modified biochar (named Z2RT400) prepared at 400℃ with a mass ratio of 2:1 (ZnCl2:rice shell) had the highest adsorption capacity for MO. Under the following conditions with a solution pH value of 4, adsorbent dosage of 10 mg, initial MO concentration of 2000 mg·L-1, and reaction time of 420 min, the maximum adsorption capacity of Z2RT400 was 1967.72 mg·g-1. When the adsorbent dosage was 80 mg, the maximum removal rate reached 99.52%. The adsorption data fitted well with the pseudo-second order kinetic model and Freundlich isotherm model, which indicates that chemical adsorption is the main adsorption mechanism and physical adsorption is the auxiliary adsorption mechanism. Therefore, the waste rice shell derived biochar can be used as a highly efficient dye adsorbent in applications such as sewage treatment.