Antiviral activity of copper contact surfaces against MS2 coliphage and hepatitis a virus.

AIMS: Viral diseases can be indirectly transmitted by contaminated non-food contact surfaces to final food products by cross-contamination. The interaction of metal surfaces and viruses, MS2 coliphage and hepatitis A virus (HAV), was investigated for strategy development in decreasing this transmission risk. METHODS AND RESULTS: MS2 deposited onto stainless-steel surface was stable but inactivated at 0.95 log10 PFU min-1 on 99.9% copper surfaces. Greater copper-inactivation of MS2 was observed in (a) simple media (phosphate buffered saline, PBS) than protein-rich media (beef extract buffer), and (b) acidic than pH ≥ 6.8 environments. Among food matrices (strawberry juices and beef broth), the greatest MS2 inactivation by copper occurred in filtered strawberry juice at pH 3.5. At a reduction of 0.17 log10 PFU min-1, HAV survived longer than MS2 on copper by FRhK-4 cell infectivity assay. CONCLUSIONS: The inactivation of virus on copper surfaces was greater in acidic viral surrounding environments and in simple PBS medium. In the same 99% PBS medium, MS2 may not be an appropriate surrogate for HAV when assessing viral inactivation on copper surfaces.

Effect of high pressure processing on migration characteristics of polypropylene used in food contact materials.

The migration of small molecular mass organic compounds from polypropylene (PP) copolymer films into food simulants during and after high pressure processing (HPP) was studied. An overlapping temperature profile was developed to isolate the pressure effect of HPP (700 MPa, 71°C, 5 min) from equivalent thermal processing (TP) at atmospheric pressure (0.1 MPa). Chloroform, toluene, methyl salicylate, and phenylcyclohexane were chosen as surrogate compounds, and were spiked into test polymer films at concentrations of 762-1152 mg kg-1 by a solvent soaking technique. Migration (w/w) of surrogate compounds from loaded PP films into Miglyol 812 (a medium-chain triglyceride mixture) and 10% ethanol was quantified by headspace GC/MS during HPP and TP, and subsequent storage at 25°C for up to 10 days. HPP significantly delayed migration of the surrogates from PP into both food simulants relative to TP. The average migrations into Miglyol after TP and HPP were 92.2-109% and 16-60.6%, respectively. Diffusion coefficients estimated by migration modelling showed a reduction of more than two orders of magnitude for all surrogate compounds under high pressure at 700 MPa (AP' = 8.0) relative to equivalent TP at 0.1 MPa (AP' = 13.1). The relative Tg increase of PP copolymer under compression at 700 MPa was estimated as Tg+94°C. For 10% ethanol, average migrations after TP and HPP were 9.3-50.9% and 8.6-22.8%, respectively. During extended storage, migration into both simulants from HPP-treated samples was initially slower than that from untreated or TP-treated films. However, after 8-24 hours of storage, the differences in percent migration of selected surrogates were not significant (p > .05) among the treated PP films. Therefore, the physical changes of PP films that occur during HPP appear to be reversible with a return to their original dimensions and diffusion properties after decompression.

Temperature and pH affect copper release kinetics from copper metal foil and commercial copperware to food simulants.

Copper (Cu) metal and alloys are used in cookware and other food contact surfaces due to their desirable properties for various applications. However, Cu metal can ionise and subsequently transfer to food and beverages under certain conditions. Here, we tested how pH and temperature affected Cu release kinetics using model systems utilising Cu metal foil and commercially available copperware. Cu foil and copperware were exposed to food simulants composed of 3% (w:w) aqueous solutions of citric acid, malic acid, acetic acid, or deionised (DI) water at temperatures ranging from 4°C to 60°C. An additional pilot experiment tested how simulated long-term cleaning affected subsequent Cu release from lined and unlined copperware to 3% citric acid. Food simulants were then analysed by ICP-MS for total Cu. After 180 min, incubation of Cu metal foil with acid-containing food simulants at 4°C resulted in Cu release ranging from 8.7 - 14.0 µg cm-2, while 21.5-38.1 µg cm-2 was released at 60°C. In contrast, Cu transfer from metal foil to DI water was relatively low, with <0.6 µg cm-2 released after 180 min at 60°C. With citric acid food simulant, lined copperware released between 0.6 and 3.0 µg Cu cm-2 over 180 min at the set temperatures, while unlined copperware released approximately 25-45 fold higher amounts of Cu (26.9-74.6 µg cm-2) over this same time period. In contrast, use of DI water food simulant resulted in Cu release of <0.1 µg cm-2 for the lined copperware and <2 µg cm-2 for the unlined type. No significant effect of simulated long-term cleaning on Cu release from copperware was observed. These data indicate that Cu release is affected by temperature and pH, and that specific steps can be taken to limit Cu metal release from food contact surfaces to foods and beverages.

Influence of Different Acids on the Transport of CdSe Quantum Dots from Polymer Nanocomposites to Food Simulants.

We fabricated polymer nanocomposites (PNCs) from low-density polyethylene and CdSe quantum dots (QDs) and used these materials to explore potential exposure after long-term storage in different acidic media that could be encountered in food contact applications. While the low-level release of QD-associated mass into all the food simulants was observed, exposure to dilute acetic acid resulted in more than double the mass transfer compared to that which occurred during exposure to dilute hydrochloric acid at the same pH. Conversely, exposure to citric acid resulted in a suppression of QD release. Permeation experiments and confocal microscopy were used to reveal mechanistic details underlying these mass-transfer phenomena. From this work, we conclude that the permeation of undissociated acid molecules into the polymer, limited by partitioning of the acids into the hydrophobic polymer, plays a larger role than pH in determining exposure to nanoparticles embedded in plastics. Although caution must be exercised when extrapolating these results to PNCs incorporating other nanofillers, these findings are significant because they undermine current thinking about the influence of pH on nanofiller release phenomena. From a regulatory standpoint, these results also support current guidance that 3% acetic acid is an acceptable acidic food simulant for PNCs fabricated from hydrophobic polymers because the other acids investigated resulted in significantly less exposure.

Effect of the local microenvironment on survival and thermal inactivation of Salmonella in low- and intermediate-moisture multi-ingredient foods.

Multi-ingredient foods having low- or intermediate-moisture characteristics may pose a special challenge to process design and validation. Ingredients of these foods can create local microenvironments that may have a distinct impact on pathogen survival and processing requirements. In this study, two model systems, each consisting of 80% commercial peanut butter (P) and 20% nonfat dry milk powder (M), were formulated to be identical in composition, but different in the source of the Salmonella contamination as originating in either the ingredient P or M. Immediately after inoculation, Salmonella showed a 2.0-log reduction when M was the contaminated ingredient compared with a 0.6-log reduction when P was the contaminated ingredient. This pattern of survival was consistent with the single-ingredient control containing only M (2.5-log reduction) or only P (0.7-log reduction), suggesting that the immediate proximity of cells is determined by the contaminated ingredient in the model system. After 5 weeks of storage, the survival rates of Salmonella in the two systems remained different, i.e.a 4- and 2-log reduction resulted in the system with M or P as the contaminated ingredient, respectively. Furthermore, thermal inactivation efficacies also differed significantly between the two systems. Fourier transform infrared spectroscopy demonstrated the nonhomogeneous distribution of water, lipid, and protein, indicating that varied local microenvironments were present and likely affected the behavior of the pathogen. The impact of the microenvironment on inactivation and survival of Salmonella was further confirmed in a butter cookie formulation in which Salmonella was inoculated via four different ingredients. This study shows that the local microenvironment in low- and intermediate-moisture foods affects Salmonella survival and thermal inactivation. The ingredient source of the contamination should be taken into account for process design and validation to ensure the safety of the product.

Static liquid permeation cell method for determining the migration parameters of low molecular weight organic compounds in polyethylene terephthalate.

The migration of low molecular weight organic compounds through polyethylene terephthalate (PET) films was determined by using a custom permeation cell assembly. Fatty food simulant (Miglyol 812) was added to the receptor chamber, while the donor chamber was filled with 1% and 10% (v/v) migrant compounds spiked in simulant. The permeation cell was maintained at 40°C, 66°C, 100°C or 121°C for up to 25 days of polymer film exposure time. Migrants in Miglyol were directly quantified without a liquid-liquid extraction step by headspace-GC-MS analysis. Experimental diffusion coefficients (DP) of toluene, benzyl alcohol, ethyl butyrate and methyl salicylate through PET film were determined. Results from Limm's diffusion model showed that the predicted DP values for PET were all greater than the experimental values. DP values predicted by Piringer's diffusion model were also greater than those determined experimentally at 66°C, 100°C and 121°C. However, Piringer's model led to the underestimation of benzyl alcohol (Áp = 3.7) and methyl salicylate (Áp = 4.0) diffusion at 40°C with its revised "upper-bound" Áp value of 3.1 at temperatures below the glass transition temperature (Tg) of PET (<70°C). This implies that input parameters of Piringer's model may need to be revised to ensure a margin of safety for consumers. On the other hand, at temperatures greater than the Tg, both models appear too conservative and unrealistic. The highest estimated Áp value from Piringer's model was 2.6 for methyl salicylate, which was much lower than the "upper-bound" Áp value of 6.4 for PET. Therefore, it may be necessary further to refine "upper-bound" Áp values for PET such that Piringer's model does not significantly underestimate or overestimate the migration of organic compounds dependent upon the temperature condition of the food contact material.

Cyclodextrin inclusion complex formation and solid-state characterization of the natural antioxidants alpha-tocopherol and quercetin.

Cyclodextrin (CD) complexation procedures are relatively simple processes, but these techniques often require very specific conditions for each individual guest molecule. Variations of the coprecipitation from aqueous solution technique were optimized for the CD complexation of the natural antioxidants alpha-tocopherol and quercetin. Solid inclusion complex products of alpha-tocopherol/beta-CD and quercetin/gamma-CD had molar ratios of 1.7:1, which were equivalent to 18.1% (w/w) alpha-tocopherol and 13.0% (w/w) quercetin. The molar reactant ratios of CD/antioxidant were optimized at 8:1 to improve the yield of complexation. The product yields of alpha-tocopherol/beta-CD and quercetin/gamma-CD complexes from their individual reactants were calculated as 24 and 21% (w/w), respectively. ATR/FT-IR, 13C CP/MAS NMR, TGA, and DSC provided evidence of antioxidant interaction with CD at the molecular level, which indicated true CD inclusion complexation in the solid state. Natural antioxidant/CD inclusion complexes may serve as novel additives in controlled-release active packaging to extend the oxidative stability of foods.

Comparison of total folate concentrations in foods determined by microbiological assay at several experienced U.S. commercial laboratories.

Analysis of total folate concentration measured by microbiological assay in a variety of foods submitted in a routine manner to experienced laboratories that regularly perform folate analysis on fee-for-service basis was evaluated. Homogenates of fresh strawberries, frozen spinach, orange juice, frozen meat and vegetable pizza, dry macaroni, and dried pinto beans were prepared and stored under conditions previously determined to maintain stability of folate content. An aliquot of each composite and of 3 certified reference materials were sent on each of 4 occasions to 4 laboratories. Results for macaroni and pizza, the only folic acid-fortified foods, had considerably lower between-laboratory variation (CV(B)) with CV(B) of 9-11% versus >45% for other foods. Mean total folate ranged from 14 to 279 microg/100 g for a mixed vegetable reference material, from 5 to 70 microg/100 g for strawberries, and from 28 to 81 microg/100 g for wholemeal flour. Only 1 laboratory reported using a tri-enzyme extraction, and all laboratories used folic acid fortified foods as internal control materials. Users of commercial total folate analysis should understand the uncertainty in values determined by microbiological assay, particularly for foods containing primarily naturally occurring folate, which may not be apparent when replicate samples are not submitted for analysis.

Diazoxide enhances adipose tissue protein kinase B activation and glucose transporter-4 expression in obese Zucker rats.

BACKGROUND: Attenuation of hyperinsulinemia in obese Zucker rats by diazoxide (DZ) enhanced insulin sensitivity and insulin-stimulated glucose uptake in isolated adipocytes. To determine if these metabolic effects are due to changes in glucose transporter (Glut)-4 gene products and intracellular signaling, we studied the effects DZ on adipose tissue Glut-4 gene products, insulin receptor substrate (IRS)-1, total and phosphorylated protein kinase B (PKB)/Akt. MATERIAL/METHODS: DZ (150 mg/kg per day) or vehicle (control) was administered to 7-week-old female obese and lean Zucker rats for 6 weeks. RESULTS: While adipose Glut-4 mRNA levels from control obese and lean rats were similar, Glut-4 protein content was 60% lower in obese than lean animals (p<0.05). DZ treatment increased mRNA in both obese (1.4 fold) and lean (1.7 fold) animals compared to controls (p<0.05), which was associated with a 3.7 fold and a 1.4 fold increase in Glut-4 protein content in DZ obese (p<0.01) and lean (p<0.05) rats, respectively. IRS-1 protein expression was lower in obese compared to lean rats (p<0.01) and was enhanced in DZ-treated obese (p<0.02) and lean (p<0.05) rats. While the PKB/Akt protein levels were similar in both strains, obese had lower p-Akt levels than lean rats (p<0.01). DZ-treated obese and lean rats had higher levels of p-Akt than their controls (p<0.05). CONCLUSIONS: Chronic suppression of hyperinsulinemia in obese Zucker rats improved intracellular insulin signaling and Glut-4 gene expression, corresponding to enhanced glucose uptake in isolated adipocytes. The discrepancy between adipose tissue Glut-4 mRNA and protein content in response to DZ treatment suggests post-transcriptional regulatory effects resulting from enhanced metabolic efficiency of insulin action.

Formation of natamycin:cyclodextrin inclusion complexes and their characterization.

Natamycin is a broad spectrum antimycotic with very low water solubility, which is used to extend the shelf life of shredded cheese products. beta-Cyclodextrin (beta-CD), hydroxypropyl beta-cyclodextrin (HP beta-CD), and gamma-cyclodextrin (gamma-CD) were found to form inclusion complexes with natamycin in aqueous solution. The increase in solubility of natamycin with added beta-CD was observed to be linear (type A(L) phase solubility diagram). The 1:1 stability constant of natamycin:beta-CD complex was estimated from its phase solubility diagram to be 1010 M(-1). The phase solubility diagrams of both gamma-CD and HP beta-CD exhibited negative deviation from linearity (type A(N) diagram) and, therefore, did not allow the estimation of binding constants. The water solubility of natamycin was increased 16-fold, 73-fold, and 152-fold with beta-CD, gamma-CD, and HP beta-CD, respectively. The natamycin:CD inclusion complexes resulted in in vitro antifungal activity nearly equivalent to that of natamycin in its free state.

Stability of natamycin and its cyclodextrin inclusion complexes in aqueous solution.

Aqueous solutions of natamycin and its beta-cyclodextrin (beta-CD), hydroxypropyl beta-cyclodextrin, and gamma-cyclodextrin (gamma-CD) inclusion complexes were completely degraded after 24 h of exposure to 1000 lx fluorescent lighting at 4 degrees C. After 14 days of storage in darkness at 4 degrees C, 92.2% of natamycin remained in active form. The natamycin:beta-CD complex and natamycin:gamma-CD complex were significantly more stable (p < 0.05) than natamycin in its free state in aqueous solutions stored in darkness at 4 degrees C. Clear poly(ethylene terephthalate) packaging with a UV light absorber allowed 85.0% of natamycin to remain after 14 days of storage under 1000 lx fluorescent lighting at 4 degrees C. Natamycin:cyclodextrin complexes can be dissociated for analysis in methanol/water/acetic acid, 60:40:5, v/v/v. Natamycin and its complexes in dissociated form were quantified by reverse phase HPLC with detection by photodiode array at 304 nm.

Chronic suppression of insulin by diazoxide alters the activities of key enzymes regulating hepatic gluconeogenesis in Zucker rats.

OBJECTIVES: Chronic attenuation of hyperinsulinemia by diazoxide (DZ), an inhibitor of glucose-mediated insulin secretion, improved insulin sensitivity and glucose tolerance and caused down-regulation of lipid metabolizing enzymes in adipose tissue and decreased the rate of weight gain in mildly hyperglycemic obese Zucker rats. Since the liver plays a central role in glucose homeostasis, we studied the effect of chronic insulin suppression on key insulin-sensitive enzymes regulating hepatic gluconeogenesis. METHODS: DZ (150 mg/kg per day) or vehicle (control) was administered to 7-week-old female obese and lean Zucker rats for a period of 4 weeks. RESULTS: DZ-treated animals showed lower fasting plasma insulin levels (P<0.001) than their controls. Plasma glucose levels were lower in DZ obese rats than in controls (P<0.001), without a significant change in DZ lean animals. DZ had no effect on glucose transporter 2 protein expression in either strain. DZ treatment resulted in lower hepatic glucokinase (P<0.001) and glucose-6-phosphatase (P<0.0001) and phosphoenolpyruvate carboxykinase (PEPCK) activities only in obese rats compared with controls (P<0.001). However, DZ-treated lean rats demonstrated higher PEPCK activity than controls (P<0.002). DZ-treated animals demonstrated enhanced hepatic glucose-6-phosphate content (P<0.01), glycogen synthase activity (P<0.0001) and glycogen content (P<0.02) compared with their controls despite increased hepatic glycogen phosphorylase a activity in these animals (P<0.02). CONCLUSIONS: Chronic suppression of hyperinsulinemia in obese Zucker rats by DZ decreased the activities of key enzymes regulating hepatic gluconeogenesis, implying that attenuation of the hyperinsulinemic state by DZ may be therapeutically beneficial.

Accelerating Scientific Discovery Through Computation and Visualization II.

This is the second in a series of articles describing a wide variety of projects at NIST that synergistically combine physical science and information science. It describes, through examples, how the Scientific Applications and Visualization Group (SAVG) at NIST has utilized high performance parallel computing, visualization, and machine learning to accelerate research. The examples include scientific collaborations in the following areas: (1) High Precision Energies for few electron atomic systems, (2) Flows of suspensions, (3) X-ray absorption, (4) Molecular dynamics of fluids, (5) Nanostructures, (6) Dendritic growth in alloys, (7) Screen saver science, (8) genetic programming.