Liquid phase exfoliation of graphite in six different animal sera and evaluation of its toxicity are reported here. Previously, we reported the exfoliation of graphene using proteins and here we extend this approach complex animal fluids. The kitchen blender with high turbulence flow gave high quality and maximum exfoliation efficiency in all sera tested, when compared to shear and ultrasonication methods. Raman spectra and electron microscopy confirmed the formation of 3-4 layer, submicron size graphene, independent of the serum used. Graphene prepared in serum was directly transferred to cell culture media without post treatments. Contrary to many reports, nanotoxicity study of this fully dispersed graphene to human embryonic kidney cells, human lung cancer cells and nematodes (C. elegans) showed no acute toxicity for up to 7 days at various doses (50-500 μg/mL). But prolonged exposure at higher doses (300-500 μg/mL, 10-15 days) showed cytotoxicity to cells (~95% death) and reproductive toxicity to C. elegans (5-10% reduction in brood size). The origin of toxicity was found to be due to the highly fragmented smaller graphene sheets (<200 nm), while the larger sheets were non-toxic (50-300 μg/mL dose). In contrast, graphene produced with sodium cholate as the mediator has been found to be cytotoxic to these cells at these dosages. We demonstrated the toxicity of liquid phase exfoliated graphene is attributed to highly fragmented fractions or non-biocompatible exfoliating agents. Thus, low toxicity graphene/serum suspensions are produced by a facile method in biological media, this approach may accelerate the much-anticipated development of graphene for biological applications.
Consumption of raw mangoes has led to multiple Salmonella-associated foodborne outbreaks in the United States. Although several studies have investigated the epiphytic fitness of Salmonella on fresh produce, there is sparse information available on the survival of Salmonella on mangoes under commercial handling and storage conditions. Hence, the objective of the study was to evaluate the survival of Salmonella on mangoes under ambient conditions simulating the mango packing house and importer facility. Further, the ability of the pathogen to adhere and attach on to the mango fructoplane was also investigated. For the attachment assays, mango skin sections were inoculated with fifty microliters of S. Newport suspension (6.5 log CFU/skin section) and minimum time required for adhesion and attachment were recorded. With the survival assays, unwaxed mangoes were spot inoculated with the Salmonella cocktail to establish approximately 4 and 6.5 log CFU/mango. The fruits were then subjected to different storage regimens simulating fruit unloading, waxing, and storage at the packing house and ripening and storage at the importer facility. Results of our study reveal that Salmonella was able to adhere on to the fructoplane immediately after contact. Further, formation of attachment structures was seen as early as 2 min following inoculation. With the survival assays, irrespective of the inoculum levels, no significant increase or decrease in pathogen population was observed when fruit were stored either at ambient (29–32°C and RH 85–95%, for 48 h), ripening (20–22°C and RH 90–95% for 9 days) or refrigerated storage (10–15°C and 85–95% for 24–48 h) conditions. Therefore, once contaminated, mangoes could serve as potential vehicles in the transmission of Salmonella along the post-harvest environment. Hence development and adoption of effective food safety measures are warranted to promote the microbiological safety of mangoes.
The gut-brain axis is thought to play a significant role in the development and manifestation of neurologic diseases. This study reports significant alterations in the brain dopamine metabolism in mice infected with C. difficile , an important pathogen that overgrows in the gut after prolonged antibiotic therapy. Such alterations in specific brain regions may have an effect on the precipitation or manifestation of neurodevelopmental disorders in humans.
This study investigated the prophylactic and therapeutic efficacies of baicalin (BC), a plant-derived flavone glycoside, in reducing the severity of Clostridioides difficile infection (CDI) in a mouse model. In the prophylactic trial, C57BL/6 mice were provided with BC (0, 11, and 22 mg/L in drinking water) from 12 days before C. difficile challenge through the end of the experiment, whereas BC administration started day 1 post challenge in the therapeutic trial. Both challenge and control groups were infected with 106 CFU/mL of hypervirulent C. difficile BAA 1803 spores or sterile PBS, and the clinical and diarrheal scores were recorded for 10 days post challenge. On day 2 post challenge, fecal and tissue samples were collected from mice prophylactically administered with BC for microbiome and histopathologic analysis. Both prophylactic and therapeutic supplementation of BC significantly reduced the severity of colonic lesions and improved CDI clinical progression and outcome compared with control (p < 0.05). Microbiome analysis revealed a significant increase in Gammaproteobacteria and reduction in the abundance of protective microbiota (Firmicutes) in antibiotic-treated and C. difficile-infected mice compared with controls (p < 0.05). However, baicalin supplementation favorably altered the microbiome composition, as revealed by an increased abundance in beneficial bacteria, especially Lachnospiraceae and Akkermansia. Our results warrant follow-up investigations on the use of BC as an adjunct to antibiotic therapy to control gut dysbiosis and reduce C. difficile infection in humans.
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