Characterization of size-differentiated airborne particulate matter collected from indoor environments of childcare facilities.

Inhalation of particulate matter (PM) present in indoor atmospheres has been associated with poor health and wellbeing of occupants. Here we report the characteristics of airborne PM collected from twenty-two air-conditioned childcare centres in Singapore. Airborne PM were collected using cascade impactors and characterized for morphology, elemental composition, endotoxin levels, ability to generate abiotic reactive oxygen species, and oxidative stress-dependent cytotoxicity in BEAS-2B cell lines. The mass concentrations of ultrafine particles (PM0.06-1) were more abundant than that of larger particles (PM1-4, PM4-20, and PM20-35 particles). PM20-35 and PM4-20 were irregularly shaped particles, PM1-4 particles had membranous flaky structures and PM0.06-1 particles were pseudo-spherical with the occasional presence of crystalline structures. Carbonaceous matter dominated PM20-35 particles, and the abundance of inorganic salts, iron and sulfur increased with decreasing PM size. Measured endotoxin levels were especially higher in PM4-20 particles. Compared to other particle size fractions, PM0.06-1 particles generated the highest ROS and were also the most potent in generating intracellular ROS in BEAS-2B cell lines. However, total mass concentrations, elemental compositions, abiotic responses, and PM collected from centres with split air-conditioning systems and no active outdoor air supply (SAC) were not statistically different compared with PM collected from centres with air conditioning with mechanical ventilation (ACMV). In conclusion, our study showed obvious distinctions in mass concentrations, morphology, elemental compositions, and cytotoxic potential of different sized particles collected from childcare centres, where the smallest particles (PM0.06-1) exhibited higher hazard potential.

Mechanistic study of the increased phototoxicity of titanium dioxide nanoparticles to Chlorella vulgaris in the presence of NOM eco-corona.

Widespread applications and release of photoactive nanoparticles (NPs) such as titanium dioxide (TiO2) into environmental matrices warrant mechanistic investigations addressing toxicity of NPs under environmentally relevant conditions. Accordingly, we investigated the effects of surface adsorbed natural organic matters (NOMs) such as humic acid, tannic acid and lignin on the band gap energy, abiotic reactive oxygen species (ROS) generation, surface chemistry and phototoxicity of TiO2 NPs. Initially, a liquid assisted grinding method was optimized to produce TiO2 NPs with a NOM layer of defined thickness for further analysis. Generally, adsorption of NOM reduced the band-gap energy of TiO2 NPs from 3.08 eV to 0.56 eV with humic acid, 1.92 eV with tannic acid and 2.48 eV with lignin. Light activated ROS generation by TiO2 NPs such as hydroxyl radicals, however, was reduced by 4, 2, 9 times in those coated with humic acid, tannic acid and lignin, respectively. This reduction in ROS despite decrease in band gap energy corroborated with the decreased surface oxygen vacancy (as revealed by X-ray Photoelectron Spectroscopy (XPS)) and quenching of ROS by surface adsorbed NOM. Despite the reduced ROS generation, the NOM-modified TiO2 NPs exhibited an increased phototoxicity to Chlorella vulgaris in comparison to pristine TiO2 NPs. Further analysis suggested that photoactivation of NOM modified TiO2 NPs releases toxic degradation products. Findings from our studies thus provide mechanistic insight into the ecotoxic potential of NOM-modified TiO2 NPs when exposed to light in the environment.

The potential of dietary nanoparticles to enhance allergenicity of milk proteins: an in vitro investigation.

In recent years, the popularity of dietary nanoparticles (NPs) in the food industry as additives has raised concerns because of the lack of knowledge about potential adverse health outcomes ensuing from the interactions of NPs with components of the food matrix and gastrointestinal system. In this study, we used a transwell culture system that consisted of human colorectal adenocarcinoma (Caco-2) cells in the apical insert and Laboratory of Allergic Diseases 2 mast cells in the basal compartment to study the effect of NPs on milk allergen delivery across the epithelial layer, mast cell responses and signaling between epithelial and mast cells in allergenic inflammation. A library of dietary particles (silicon dioxide NPs, titanium dioxide NPs and silver NPs) that varied in particle size, surface chemistry and crystal structures with or without pre-exposure to milk was used in this investigation. Milk-interacted particles were found to acquire surface corona and increased the bioavailability of milk allergens (casein and ß-lactoglobulin) across the intestinal epithelial layer. The signaling between epithelial cells and mast cells resulted in significant changes in the early phase and late-phase activation of the mast cells. This study suggested that antigen challenge in mast cells with the presence of dietary NPs may cause the transition of allergic responses from an immunoglobulin E (IgE)-dependent mechanism to a mixed mechanism (both IgE-dependent and IgE-independent mechanisms).

Genomic and phenotypic profiling of Staphylococcus aureus isolates from bovine mastitis for antibiotic resistance and intestinal infectivity.

BACKGROUND: Staphylococcus aureus is one of the prevalent etiological agents of contagious bovine mastitis, causing a significant economic burden on the global dairy industry. Given the emergence of antibiotic resistance (ABR) and possible zoonotic spillovers, S aureus from mastitic cattle pose threat to both veterinary and public health. Therefore, assessment of their ABR status and pathogenic translation in human infection models is crucial. RESULTS: In this study, 43 S. aureus isolates associated with bovine mastitis obtained from four different Canadian provinces (Alberta, Ontario, Quebec, and Atlantic provinces) were tested for ABR and virulence through phenotypic and genotypic profiling. All 43 isolates exhibited crucial virulence characteristics such as hemolysis, and biofilm formation, and six isolates from ST151, ST352, and ST8 categories showed ABR. Genes associated with ABR (tetK, tetM, aac6', norA, norB, lmrS, blaR, blaZ, etc.), toxin production (hla, hlab, lukD, etc.), adherence (fmbA, fnbB, clfA, clfB, icaABCD, etc.), and host immune invasion (spa, sbi, cap, adsA, etc.) were identified by analyzing whole-genome sequences. Although none of the isolates possessed human adaptation genes, both groups of ABR and antibiotic-susceptible isolates demonstrated intracellular invasion, colonization, infection, and death of human intestinal epithelial cells (Caco-2), and Caenorhabditis elegans. Notably, the susceptibilities of S. aureus towards antibiotics such as streptomycin, kanamycin, and ampicillin were altered when the bacteria were internalized in Caco-2 cells and C. elegans. Meanwhile, tetracycline, chloramphenicol, and ceftiofur were comparatively more effective with ≤ 2.5 log10 reductions of intracellular S. aureus. CONCLUSIONS: This study demonstrated the potential of S. aureus isolated from mastitis cows to possess virulence characteristics enabling invasion of intestinal cells thus calling for developing therapeutics capable of targeting drug-resistant intracellular pathogens for effective disease management.

Transcriptomic points of departure calculated from human intestinal cells exposed to dietary nanoparticles.

Use of nanoparticles (NPs) in the food industry raises dietary health concerns. Assessing dietary NPs remains challenged due the vast number of products and the resource-intensive nature of toxicity testing. Advancements in high-throughput transcriptomics, coupled with benchmark dose (BMD) analysis are poised to modernize chemical safety assessments. The study objective was to derive transcriptomic point of departure (tPOD) values for common dietary NPs through dose-response analysis of 3'RNA-sequencing data. Two intestinal cell lines (Caco-2, HIEC-6) were exposed to 9 forms of Ag, SiO2, and TiO2, and expression of L1000 landmark genes was characterized. In Caco-2 cells, tPODmode concentrations were 0.4-0.6, 21-32, and 17-59 ppm for NPs of Ag, SiO2, and TiO2, respectively; in HIEC-6 cells, the respective tPOD values were 6-7, 7-9, and 3-13 ppm. Pathway BMDs across cases identified, for example, osteoclast and Th1/Th2 cell differentiation, and cell cycle, signaling, and senescence pathways. In all cases, the tPOD and pathway BMD values were lower than concentrations associated with cellular changes (e.g., generation of reactive oxygen species and proinflammatory cytokines, and cytotoxicity). These results demonstrate that transcriptomics dose-response analysis using in vitro models can help to increase understanding of a NP's mechanisms of action and derive quantitative information for dietary risk assessment.

Hazard Profiling of Commercially Relevant Quantum Dot Components Revealed Synergistic Interactions between Heavy Metals and Polymers.

Commercially used quantum dots (QDs) exemplify complex nanomaterials with multiple components, though little is known about the type of interactions between these components in determining the overall toxicity of this material. We synthesized and characterized a functional QD (CdSe/ZnS_P&E) that was identical in structure and composition to a patented and commercially applied QD and the combinations of its components (CdSe, CdSe/ZnS, ZnS, CdSe_P&E, ZnS_P&E, and P&E). Cells exposed to incremental concentrations of these materials were investigated for cell viability and cellular perturbations, contributing to a final common pathway of cell death using high-content screening assays in model human intestinal epithelial cells (HIEC-6). The concentrations that resulted in a loss of 20% cell viability (EC20 values) for each tested component were used for estimating the combination index (CI) to evaluate synergistic or antagonistic effects between the components. Complete QD (core/shell-polymer) showed the highest toxic potential due to synergistic interactions between core and surface functional groups. The cationic polymer coating enhanced cellular uptake of the QD, ensuing lysosome acidification and release of heavy metal ions to the intracellular milieu, and caused oxidative stress and cytotoxicity. Overall, this study advances our understanding of the collective contribution of individual components of a functional QD toward its toxic potential and emphasizes the need to study multilayered nanomaterials in their entirety for hazard characterization.

Nanoparticle-Enabled Combination Therapy Showed Superior Activity against Multi-Drug Resistant Bacterial Pathogens in Comparison to Free Drugs.

The emergence of multidrug-resistant (MDR) bacterial pathogens in farm animals and their zoonotic spread is a concern to both animal agriculture and public health. Apart from antimicrobial resistance (AMR), bacterial pathogens from the genera of Salmonella and Staphylococcus take refuge inside host cells, thereby demanding intervention strategies that can eliminate intracellular MDR pathogens. In this study, seven clinical isolates of Salmonella and Staphylococcus from swine farms were characterized for antibiotic (n = 24) resistance, resistance mechanisms, and virulence characteristics. All isolates showed resistance to one or more antibiotics and S. enterica ser. Typhimurium isolate had the highest resistance to the panel of antibiotics tested. Major resistance mechanisms identified were efflux pump and beta-lactamase enzyme activities. Staphylococcus isolates showed complete hemolysis and strong biofilm formation, while Salmonella isolates caused partial hemolysis, but showed no or weak biofilm formation. MDR isolates of S. aureus M12 and S. enterica ser. Typhimurium bacteria were subsequently tested against combinations of antibiotics and potentiating adjuvants for improved antibacterial efficacy using a checkerboard assay, and their fractional inhibitory concentration index (FICI) was calculated. A combination of chitosan and silica nanoparticles containing tetracycline (TET) and efflux pump inhibitor chlorpromazine (CPZ), respectively, was characterized for physicochemical properties and effectiveness against MDR Salmonella enterica ser. Typhimurium isolate. This combination of nano-encapsulated drugs improved the antibacterial efficacy by inhibiting AMR mechanisms (efflux activity, beta-lactamase enzyme activity, and hydrogen sulfide (H2S) production) and reducing intracellular pathogen load by 83.02 ± 14.35%. In conclusion, this study sheds light on the promising applicability of nanoparticle-enabled combination therapy to combat multidrug-resistant pathogens encountered in animal agriculture.

Light activation of gold nanorods but not gold nanospheres enhance antibacterial effect through photodynamic and photothermal mechanisms.

Plasmonic nanomaterials of gold and silver have been reported to have antibacterial effect. In this study, three gold nanomaterials (NMs) of different aspect rations (Gold nanospheres (AuNSs, aspect ratio 1), and two gold nanorods (AuNRs636, aspect ratio 2.79; AuNRs772, aspect ratio 3.42)) and silver nanoparticles (AgNPs) were synthesized, characterized and the effect of incandescent light on their antibacterial properties were examined. Bacterial inactivation during photoinactivation of nanomaterials and antibacterial mechanisms (biotic ROS, membrane potential, membrane damage) were investigated using Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Salmonella enterica serovar Typhimurium, and methicillin-resistant S. aureus. The results indicated that AuNSs had no antibacterial activity in the tested concentration (0.49-250 µg/mL), while AuNR636 and AuNRs772 showed significant bactericidal effect on all tested bacteria. Notably, AuNRs636 presented higher antibacterial effect than AuNRs772, which could result from higher surface reactivity of AuNRs636 owing to higher dangling bonds. Further studies showed that AuNRs but not AuNSs generated hydroxyl radicals (·OH) (photodynamic effect) and photothermal effect when exposed to incandescent light. The combined photodynamic and photothermal effect resulted in bacterial inactivation through cell membrane damage, lowering of cell membrane potential and DNA degradation. In summary, this investigation showed that Au NRs but not Au NSs exhibit photodynamic and photothermal effects suggesting the potential of fabricating material surfaces with Au NRs for photoactivated bacterial inactivation.

Composite of Layered Double Hydroxide with Casein and Carboxymethylcellulose as a White Pigment for Food Application.

Titanium dioxide (TiO2) is commonly used in food, cosmetic, and pharmaceutical industries as a white pigment due to its extraordinary light scattering properties and high refractive index. However, as evidenced from recent reports, there are overriding concerns about the safety of nanoparticles of TiO2. As an alternative to TiO2, Mg-Al layered double hydroxide (LDH) and their composite containing casein and carboxymethyl cellulose (CMC) were synthesized using wet chemistry and compared with currently used materials (food grade TiO2 (E171), rice starch, and silicon dioxide (E551)) for its potential application as a white pigment. These particles were characterized for their size and shape (Transmission Electron Microscopy), crystallographic structure (X-Ray Diffraction), agglomeration behavior and surface charge (Dynamic Light Scattering), surface chemistry (Fourier Transform Infrared Spectroscopy), transmittance (UV-VIS spectroscopy), masking power, and cytotoxicity. Our results showed the formation of typical layered double hydroxide with flower-like morphology which was restructured into pseudo-spheres after casein intercalation. Transmittance measurement showed that LDH composites had better performance than pristine LDH, and the aqueous suspension was heat and pH resistant. While its masking power was not on a par with E171, the composite of LDH was superior to current alternatives such as rice starch and E551. Sustainability score obtained by MATLAB® based comparison for price, safety, and performance showed that LDH composite was better than any of the compared materials, highlighting its potential as a white pigment for applications in food.

Hazard profiling of a combinatorial library of zinc oxide nanoparticles: Ameliorating light and dark toxicity through surface passivation.

Zinc oxide (ZnO) is one of the high-volume production nanoparticles (NPs) currently used in a wide range of consumer and industrial goods. The inevitable seepage into environmental matrices and the photoactive nature of ZnO NPs warrants hazard profiling under environmentally related conditions. In this paper, the influence of simulated solar light (SSL) on dissolution behaviour and phototoxicity of ZnO NPs was studied using a combinatorial library of ZnO NPs with different sizes, surface coatings, dopant chemistry, and aspect ratios in a fish cell line (BF2) and zebrafish embryos. Generally, the cytotoxicity and embryo mortality increased when exposed concomitantly to SSL and ZnO NPs. The increase in toxic potential of ZnO NPs during SSL exposure concurred with release of Zn ions and ROS generation. Surface modification of NPs with poly(methacrylic acid) (PMAA), silica or serum coating decreased toxicity and ZnO with serum coating was the only NP that had no significant effect on any of the cytotoxicity parameters when tested under both dark and SSL conditions. Results from our study show that exposure to light could increase the toxic potential of ZnO NPs to environmental lifeforms and mitigation of ZnO NP toxicity is possible through modifying the surface chemistry.

Inorganic food additive nanomaterials alter the allergenicity of milk proteins.

While inorganic nanomaterials are copiously incorporated in food products, their impact on the allergenicity food proteins is largely unknown. This study analyzed the effect of widely used food additive nanomaterials (silica and titania) on the antigenicity and allergenicity of milk proteins (ß-lactoglobulin and casein) and skimmed milk. Changes in the antigenicity of milk proteins in the presence of dietary nanomaterials were identified using an indirect-ELISA assay, while the change in allergenicity was studied using mast cell (LAD2) sensitized using allergic human sera. Results showed an enhancement in the allergenicity of milk proteins/skimmed milk interacted with particles (both silica and titania). Similarly, mast cell degranulation (a proxy for allergenicity) was higher when exposed to particle interacted skim milk where nanomaterials of titania showed the highest effect, and this tendency was retained even after subjecting to simulated gut digestion. Particles induced alterations in the structure of milk proteins, as evidenced by our studies, are reasoned to expose epitopes that increase allergenicity of milk proteins.

Protein-biomolecule interactions play a major role in shaping corona proteome: studies on milk interacted dietary particles.

Despite the significance of surface absorbed proteins in determining the biological identity of nanoparticles (NPs) entering the human body, little is known about the surface corona and factors that shape their formation on dietary particles used as food additives. In this study, food grade NPs of silica and titania and their food additive counterparts (E551 and E171) were interacted with milk proteins or with skimmed milk and the levels of protein adsorption were quantified. Characteristics of proteins correlating with their level of adsorption to NPs were determined using partial least squares regression analysis. Results from individual protein-particle interactions revealed the significance of factors such as zeta potential, hydrophobicity and hydrodynamic size of particles, and protein characteristics such as the number of beta strands, isoelectric points, the number of amino acid units (Ile, Tyr, Ala, Gly, Pro, Asp, and Arg), and phosphorylation sites on their adsorption to particles. Similar regression analysis was performed to identify the characteristics of twenty abundant and enriched proteins (identified using LC-MS/MS analysis) for their association with the surface corona of milk-interacted particles. Contrary to individual protein-particle interactions, protein characteristics such as helices, turns, protein structures, disulfide bonds, the number of amino acid units (Cys, Met, Leu, and Trp), and Fe binding sites were significant for their association with the surface corona of milk interacted particles. This difference in factors identified from individual proteins and milk interacted particles suggested possible interactions of proteins with surface adsorbed biomolecules as revealed by scanning transmission X-ray microscopy and other biochemical assays.

Prevalence and mechanisms of antibiotic resistance in Escherichia coli isolated from mastitic dairy cattle in Canada.

BACKGROUND: Bovine mastitis is the most common infectious disease in dairy cattle with major economic implications for the dairy industry worldwide. Continuous monitoring for the emergence of antimicrobial resistance (AMR) among bacterial isolates from dairy farms is vital not only for animal husbandry but also for public health. METHODS: In this study, the prevalence of AMR in 113 Escherichia coli isolates from cases of bovine clinical mastitis in Canada was investigated. Kirby-Bauer disk diffusion test with 18 antibiotics and microdilution method with 3 heavy metals (copper, zinc, and silver) was performed to determine the antibiotic and heavy-metal susceptibility. Resistant strains were assessed for efflux and ß-lactamase activities besides assessing biofilm formation and hemolysis. Whole-genome sequences for each of the isolates were examined to detect the presence of genes corresponding to the observed AMR and virulence factors. RESULTS: Phenotypic analysis revealed that 32 isolates were resistant to one or more antibiotics and 107 showed resistance against at least one heavy metal. Quinolones and silver were the most efficient against the tested isolates. Among the AMR isolates, AcrAB-TolC efflux activity and ß-lactamase enzyme activities were detected in 13 and 14 isolates, respectively. All isolates produced biofilm but with different capacities, and 33 isolates showed α-hemolysin activity. A positive correlation (Pearson r = + 0.89) between efflux pump activity and quantity of biofilm was observed. Genes associated with aggregation, adhesion, cyclic di-GMP, quorum sensing were detected in the AMR isolates corroborating phenotype observations. CONCLUSIONS: This investigation showed the prevalence of AMR in E. coli isolates from bovine clinical mastitis. The results also suggest the inadequacy of antimicrobials with a single mode of action to curtail AMR bacteria with multiple mechanisms of resistance and virulence factors. Therefore, it calls for combinatorial therapy for the effective management of AMR infections in dairy farms and combats its potential transmission to the food supply chain through the milk and dairy products.

Characterizing the effects of titanium dioxide and silver nanoparticles released from painted surfaces due to weathering on zebrafish (Danio rerio).

Silver (nAg) and titanium dioxide nanoparticles (nTiO2) are common engineered nanoparticles (ENPs) added into paint for their antimicrobial and whitening properties, respectively. Weathering of outdoor painted surfaces can release such ENPs, though little is known about the potential effects of released ENPs on aquatic species. The objective of this study was to characterize the toxicity of nAg and nTiO2 released from painted panels using fish liver cells (CRL2643) and zebrafish embryos (OECD 236 embryotoxicity test). Cells and embryos were exposed to suspensions of pristine nAg or nTiO2, panels (unpainted or painted with nAg or nTiO2) or base paint, after sonication. Cell viability and gene expression were assessed using resazurin assay and qPCR, respectively, while embryo mortality and deformities were scored visually via microscopic examination. In the cell studies, both paint-released nanoparticles did not affect viability, but paint-released nAg resulted in differential expression of a few genes including gclc and ncf1. In embryos, paint-released nAg increased mortality and incidence of deformities, whereas paint-released nTiO2 resulted in differential expression of several genes including gclc, ncf1, txnrd1, gpx1b, and cyp1c1 but without major phenotypic abnormalities. Comparing the two types of exposures, paint-released exposures affected both molecular (gene expression) and apical (embryotoxicity) endpoints, while pristine exposures affected the expression of some genes but had no apical effects. The differing effects of paint-released and pristine nanoparticle exposures suggest that further research is needed to further understand how paint coatings (and the products of their weathering and aging) may influence nanoparticle toxicity to aquatic organisms.

Dietary nanoparticles compromise epithelial integrity and enhance translocation and antigenicity of milk proteins: An in vitro investigation.

Nanoparticles (NPs) are increasingly being used in the food sector, yet little is known about the potential health risks associated with oral exposure to dietary NPs. In this study, the most widely used NPs in food industry including food grade silicon dioxide (SiO2), titanium dioxide (TiO2) and silver (Ag), along with their non-food grade and bulk counterparts, are characterized for physicochemical properties and molecular, cellular, and intracellular effects on human intestinal epithelial cells (Caco-2 and HIEC-6). Silver NPs are the most cytotoxic and induce significant cellular changes in oxidative stress, Ca2+ flux and mitochondria function, leading to cellular junction disruption at the lowest exposure concentration. At higher testing concentrations, NPs but not microparticles of SiO2 and TiO2 cause sublethal cellular responses and remodel tight junctions without impairing epithelial integrity. To relate the cellular results to key events in GI disorder progression, NPs are exposed to an in vitro co-culture model for cow's milk allergy comprised of Caco-2 and allergy sera-primed mast cells (LUVA). All particle treatments increase the allergen delivery across intestinal epithelium and subsequent allergy responses. Overall, the study has identified a particle-dependent alteration in intestinal epithelium and highlighted potential safety concerns of dietary NPs.

Personal level exposure and hazard potential of particulate matter during haze and non-haze periods in Singapore.

Severe haze episodes originating from biomass burning are common in Southeast Asia. However, there is a paucity of data on the personal exposure and characteristics of Particulate Matter (PM) present in ambient air during haze and non-haze periods. Aims of this study were to monitor 24 h ambulatory exposure to PM among school children in Singapore; characterize haze and non-haze PM for their physicochemical properties, cytotoxicity and inflammatory potential, using bronchial epithelial cell culture model (BEAS-2B). Forty-six children had ambulatory PM exposure monitored using portable Aethalometer and their hourly activity recorded. The mean (±SE) PM exposure on a typical school day was 3343 (±174.4) ng/m3/min. Higher PM exposure was observed during haze periods and during commuting to and from the school. Characterization of PM collected showed a drastic increase in the proportion of ultrafine particle (UFP) in haze PM. These PM fraction showed higher level of sulphur, potassium and trace metals in comparison to those collected during non-haze periods. Dose dependent increases in abiotic reactive oxygen species generation, activation of NF-κB and cytotoxicity were observed for both haze and non-haze PM. Generally, haze PM induced significantly higher release of IL-6, IL-8 and TNFα by BEAS-2B cells in comparison to non-haze PM. In summary, this study provides experimental evidence for higher PM exposure during haze period which has the potential to elicit oxidative stress and pro-inflammatory cytokine release from airway epithelial cells.

Enhancing the Bioavailability of Silver Through Nanotechnology Approaches Could Overcome Efflux Pump Mediated Silver Resistance in Methicillin Resistant Staphylococcus aureus.

While the wide-spectrum antimicrobial properties and stability of silver nanomaterials have been copiously utilized in many medical and consumer products, we found that Methicillin Resistant Staphylococcus aureus (MRSA) is less susceptible to silver in comparison to Methicillin Sensitive Staphylococcus aureus (MSSA). Pre-exposure of MRSA to sub-lethal concentrations of AgNO3 caused 2.5-fold increase in LD50 of silver suggesting an inducible resistance mechanism. Studies involving gene expression profiling and efflux pump blockers showed the induction of P-type efflux pumps (Cop A, Cop Z and Nor B) as the principle mechanism conferring silver resistance in MRSA. Chlorpromazine-an efflux pump blocker increased sensitivity of MRSA to silver. Leveraging on these observations, silver resistance in MRSA was circumvented by enhancing the bioavailability of silver by cationic functioning of silver nanoparticles or by co-delivering silver together with chlorpromazine. Atomic Force Microscopy showed that poly-ethylene imine (PEI) functionalized silver nanoparticles adhere to bacterial cells which was found to increase the bioavailability, membrane rupture and cell death. The strategy of co-delivery of AgNO3 and chlorpromazine using chitosan-functionalized wormhole silica nanoparticles caused 12 log reduction in bacterial count which was 1000 times higher than bacterial reduction by AgNO3 alone. In short, these studies showed that circumventing antimicrobial resistance in pathogenic bacteria is possible by designed silver nanotechnology.

A Comparative Analysis of Different Grades of Silica Particles and Temperature Variants of Food-Grade Silica Nanoparticles for Their Physicochemical Properties and Effect on Trypsin.

While silica particles are used extensively in food products, different grades and temperature variants of silica particles have not been compared for their physiochemical and biological properties. Different grades of silica (food-grade nanoparticles (FG-NPs), nonfood-grade nanoparticles (NFG-NPs), and food-grade micron particles (FG-MPs)) and the temperature variants generated by exposing FG-NPs to wet heating, dry heating, and refrigeration were compared for their physicochemical properties and interaction with trypsin. FG-NPs were similar to NFG-NPs and FG-MPs in their elemental composition and amorphous nature but had relatively less branched and ring siloxane groups than the latter ones. There were subtle but noticeable changes in the agglomeration behavior and relative abundance of different silica groups in FG-NPs exposed to food-handling temperatures. Secondary structure and function of trypsin were negatively impacted by FG-NPs and their temperature variants. Silica particles showed a "mixed-type inhibition" of trypsin resulting in partial digestion of bovine serum albumin. In conclusion, our studies showed differences in the surface chemistry of different grades of silica particles and temperature variants of FG-NPs and their negative impact on the structure and function of trypsin.

New Trends in the Microencapsulation of Functional Fatty Acid-Rich Oils Using Transglutaminase Catalyzed Crosslinking.

Preparing stable protein-based microcapsules containing functional fatty acids and oils for food applications has been a big challenge. However, recent advances with transglutaminase (TGase) enzyme as an effective protein cross-linker could provide workable solutions for the encapsulation of omega-3 and omega-6 fatty acids without compromising their targeted release and their biological and physicochemical characteristics. The recent and available literature related to the microencapsulation techniques, physical and oxidative properties, and core retention and release mechanisms of TGase-crosslinked microcapsules entrapping edible oils were reviewed. The effects of factors involved in microencapsulation processes, on the efficiency and quality of the produced innovative microcapsules were also discussed and highlighted. A brief focus has been finally addressed to new insights and additional knowledge on micro- and nanoencapsulation of lipophilic food-grade ingredients by TGase-induced gelation. Two dominant microencapsulation methods for fish, vegetable, and essential oils by TGase-crosslinking are complex coacervation and emulsion-based spray drying. The developed spherical particles (<100 µm) with some wrinkles and smooth surfaces showed an excellent encapsulation efficiency and yield. A negligible release rate and a substantial retention level can result for different lipid-based cores covered by TGase-crosslinked proteins during the oral digestion and storage. A significant structural, thermal and oxidative stability for edible oils-loaded microcapsules in the presence of TGase can be also obtained.