High-Pressure Processing of Fruit Smoothies Enriched with Dietary Fiber from Carrot Discards: Effects on the Contents and Bioaccessibilities of Carotenoids and Vitamin E.

The effects of high-pressure processing (HPP) (450 MPa/600 MPa/3 min) on the carotenoid and vitamin E contents of smoothies made from strawberry, orange juice, banana and apple, and the same smoothies enriched with dietary fiber from discarded carrots were compared. The contents and bioaccessibilities of these compounds were also evaluated over the course of 28 days at 4 °C. The application of HPP in the formulations significantly increased the contents of ß-cryptoxanthin, α-carotene and ß-carotene and retained the contents of lutein, zeaxanthin and vitamin E compared to untreated samples. A decreasing trend in the content of each compound was observed with an increase in storage time. The application of HPP initially led to reductions in the bioaccessibility of individual compounds. However, overall, during storage, there was an increase in bioaccessibility. This suggests that HPP influences cell structure, favoring compound release and micelle formation. HPP is a sustainable method that preserves or enhances carotenoid extractability in ready-to-drink fruit beverages. Furthermore, the incorporation of dietary fiber from carrot processing discards supports circular economy practices and enhances the health potential of the product.

High-Pressure Processing of Kale: Effects on the Extractability, In Vitro Bioaccessibility of Carotenoids & Vitamin E and the Lipophilic Antioxidant Capacity.

High pressure processing (HPP) represents a non-thermal preservation technique for the gentle treatment of food products. Information about the impact of HPP on lipophilic food ingredients (e.g., carotenoids, vitamin E) is still limited in more complex matrices such as kale. Both the variation of pressure levels (200-600 MPa) and different holding times (5-40 min) served as HPP parameters. Whereas a slightly decreasing solvent extractability mostly correlated with increasing pressure regimes; the extension of holding times resulted in elevated extract concentrations, particularly at high-pressures up to 600 MPa. Surprisingly, slightly increasing bioaccessibility correlated with both elevated pressures and extended holding times, indicating matrix-dependent processes during in vitro digestion, compared to results of extractability. Moreover, the verification of syringe filters for digest filtration resulted in the highest relative recoveries using cellulose acetate and polyvinylidene difluoride membranes. The α-tocopherol equivalent antioxidant capacity (αTEAC) and oxygen radical antioxidant capacity (ORAC) assays of treated kale samples, chopped larger in size, showed increased antioxidant capacities, regarding elevated pressures and extended holding times. Consequently, one may conclude that HPP was confirmed as a gentle treatment technique for lipophilic micronutrients in kale. Nevertheless, it was indicated that sample pre-treatments could affect HP-related processes in food matrices prior to and possibly after HPP.

Environmental impact of high-value gold scrap recycling.

PURPOSE: The gold routes satisfying the global gold supply are mining (74%), recycling of high-value gold (23%), and electronic scraps (3%). Besides its applications in the investment, jewelry, and industrial sector, gold also has a bad image. The gold production in industrial as well as artisanal and small-scale mines creates negative impacts such as resource depletion, extensive chemical use, toxic emissions, high energy consumption, and social concerns that are of great importance. On the other hand, almost all gold is recycled and has historically always been. In common life cycle assessment (LCA) databases, there is no data on recycling of high-value gold available. This article attempts to answer the question what the ecological benefits of this recycling are. METHOD: In this study, we were able to collect process data on the most commonly used high-value gold scrap recycling process, the aqua regia method, from several state-of-the-art German refineries. With this data, life cycle inventories were created and a life cycle model was produced to finally generate life cycle impacts of high-value gold scrap recycling. RESULTS: This study contains the corresponding inventories and thus enables other interested parties to use these processes for their own LCA studies. The results show that high-value gold scrap recycling has a considerably lower environmental impact than electronic gold scrap recycling and mining. For example, high-value gold scrap recycling in Germany results in a cumulative energy demand (CED) of 820 MJ and a global warming potential (GWP) of 53 kg-CO2-Eq. per kg gold. In comparison, common datasets indicate CED and GWP levels of nearly 8 GJ and 1 t-CO2-Eq. per kg gold, respectively, for electronic scrap recycling and levels of 240 GJ and 16 t-CO2-Eq. per kg gold, respectively, for mining. CONCLUSION: The results show that buying gold from precious metal recycling facilities with high technological standards and a reliable origin of the recycling material is about 300 times better than primary production.

Discrete-Point Analysis of the Energy Demand of Primary versus Secondary Metal Production.

The metal industry consumes large amounts of energy and contributes significantly, up to 10%, to global greenhouse gas (GHG) emissions. Recycling is commonly included among the most viable options for mitigating the climate forcing of metal production by replacing primary production. However, the recycling rates of metals are still incomplete and, in particular, do not exist for most specialty metals. Our empirical analysis of 48 metals shows that their recycling is mainly impeded by their low concentrations. In many cases, the metal concentration in end-of-life products is lower than that in natural ores. This phenomenon inevitably raises the question of the extent to which recycling can be conducted without losing its mitigating effects on climate change. We answer this question for two example metals, tantalum and copper, within the scope of Germany, a leader in recycling. For tantalum, the results show that a further increase in the end-of-life recycling rate (EOL-RR) could contribute to minimizing the overall energy consumption and GHG emissions, despite its low concentrations in end-of-life products. The energy requirements for recycling copper from end-of-life products already reach the magnitude of those for primary production. A further increase in EOL-RR must be examined in detail to ensure mitigating effects on climate change.

GC-ECNICI-MS analysis of S-nitrosothiols and nitroprusside after treatment with aqueous sulphide (S2-) and derivatization with pentafluorobenzyl bromide: Evidence of S-transnitrosylation and formation of nitrite and nitrate.

A GC-MS method is reported for the quantitative analysis of S-nitrosothiols (RSNO) derived from endogenous low- and high-molecular mass thiols (RSH) including hemoglobin, cysteine, glutathione, N-acetylcysteine, and the exogenous N-acetylcysteine ethyl ester. The method is based on the conversion of RSNO to nitrite by aqueous Na2S (S2-). 15N-Labelled analogs (RS15NO) or 15N-labelled nitrite and nitrate were used as internal standards. The nitrite (14NO2- and 15NO2-) and nitrate (O14NO2- and O15NO2- anions were derivatised by pentafluorobenzyl (PFB) bromide (PFB-Br) in aqueous acetone and their PFB derivatives were separated by gas chromatography. After electron-capture negative-ion chemical ionization, the anions were separated by mass spectrometry and detected by selected-ion monitoring of m/z 46 for 14NO2-, m/z 47 for 15NO2-, m/z 62 for O14NO2-, and m/z 63 for O15NO2-. The expected thionitrites (-S14NO and -S15NO) were not detected, suggesting that they are intermediates and rapidly exchange their S by O from water, presumably prior to PFB-Br derivatization. The reaction of S2- with RSNO and sodium nitroprusside (SNP) resulted in the formation of nitrite and nitrate as the major and minor reaction products, respectively. The novel Na2S procedure was compared with established procedures based on the use of aqueous HgCl2 or cysteine/Cu2+ reagents to convert the S-nitroso group to nitrite. Our results provide evidence for an equilibrium S-transnitrosylation reaction between S2- with RSNO in buffered solutions of neutral pH. Use of Na2S in molar excess over RSNO shifts this reaction to the right, thus allowing almost complete conversion of RSNO to nitrite and nitrate. The Na2S procedure should be useful for the quantitative determination of RSNO as nitrite and nitrate after PFB-Br derivatization and GC-MS analysis. The Na2S procedure may also contribute to explore the complex reactions of S2- with RSNO, SNP and other NO-containing compounds.

Evidence by chromatography and mass spectrometry that inorganic nitrite induces S-glutathionylation of hemoglobin in human red blood cells.

Previously we found by HPLC with fluorescence detection that inorganic nitrite induces oxidation of glutathione (GSH) to its disulfide (GSSG) in intact and more abundantly in lyzed red blood cells (RBCs) from healthy humans. In the present work, we performed MS-based protein analysis and observed that nitrite (range, 0-20mM) induces formation of S-glutathionyl hemoglobin (HbSSG) at cysteine (Cys) ß93 and ß112 of oxyhemoglobin (HbO2) in lyzed human RBCs (range, 6-8mM HbO2). Hemoglobin species were isolated from incubation mixtures of nitrite in lyzed RBCs by ultrafiltration or affinity chromatography and analyzed by HPLC and LC-MS/MS. The mechanism likely involves inhibition of catalase activity by nitrite (IC50, 9 µM), which allows H2O2 to accumulate and oxidize Cys moieties of oxyhemoglobin and erythrocytic GSH to form HbSSG in addition to GSSG. In freshly prepared hemolysate samples, nitrite induced release of superoxide and molecular oxygen. In the presence of paracetamol and nitrite in hemolysate samples, 3-nitro-paracetamol was detected. Nitrite also induced S-nitroso hemoglobin (HbSNO) formation in low yield (i.e., 0.1%). Synthetic cysteine (Cys), glutathione (GSH), N-acetylcysteine (NAC) and N-acetylcysteine ethyl ester (NACET) inhibited nitrite-induced modifications of oxyhemoglobin including methemoglobin, HbSSG (CysSH >> NACET >> GSH ≈ NAC; thiol concentration, 50 µM) and HbSNO. Nitrite-induced oxidative modifications may alter physiological hemoglobin functions and may require alternative treatments for conditions associated with oxidized hemoglobin like in nitrite-induced methemoglobinemia. Accumulation of soluble Cys in RBCs via oral administration of NACET could be a new promising strategy to prevent nitrite-induced methemoglobinemia by nitrite and other oxidants.

Intuitive steering assistance in critical understeer situations.

OBJECTIVE: To develop and verify a driver assistance function, working on the electric power steering of passenger cars, to support the driver on handling critical understeer situations. The main objectives of the so-called understeer assistance are reinforcing the driver's awareness of the driving conditions and giving support to handle the situation correctly without inducing irritation by abnormal steering behavior. METHODS: The system was designed in consideration of psychological aspects of human decision making while operating a vehicle in unfamiliar understeer situations. Using a comparison of vehicle dynamics with a reference model computed in the car, the level of understeer is calculated. Depending on the understeer level, the steering wheel restoring torque is increased while a vibration is applied to the steering wheel at the same time. To verify the achievement of the objectives, the standard steering system is compared to the developed understeer assistance in an active driving study on a test track. Not only objective measurement data but also subjective ratings delivered by 63 unbiased participants were used. RESULTS: The subjects follow the offered steering recommendation by steering less when the assistance function is activated (Δγmax = 44.38°, p = 2.5°·10⁻³%). In the sequel, an enhanced vehicle reaction arises that is validated by analyzing the achieved maximum lateral offset (ΔSy,res = 0.16m, p = 0.07%). In addition, the evaluation of subjective ratings clearly indicates a better awareness of the understeer situation with the assistance function (χ= +0.44, p = 0.89%). Furthermore the subjects rate the understeer assistance better than the standard steering system (χ= +0.43, p = 3.03%). CONCLUSIONS: By measuring vehicle data and eliciting subjective opinions of the participants, the effectiveness regarding an improved handling of an understeering vehicle as well as the acceptance of the understeer assistance by the driver is confirmed. Larger subject groups and more realistic boundary conditions should be considered for additional evaluations. The approach of using standard vehicle hardware to adapt the vehicle to specific driving situations under consideration of psychological aspects of human decision making should be investigated further.

UPLC-MS/MS measurement of S-nitrosoglutathione (GSNO) in human plasma solves the S-nitrosothiol concentration enigma.

We developed and validated a fast UPLC-MS/MS method with positive electrospray ionization (ESI+) for the quantitative determination of S-nitrosoglutathione (GSNO) in human plasma. We used a published protocol for the inactivation of plasma γ-glutamyltransferase (γGT) activity by using the γGT transition inhibitor serine/borate and the chelator EDTA for the stabilization of GSNO, and N-ethylmaleimide (NEM) to block SH groups and to avoid S-transnitrosylation reactions which may diminish GSNO concentration. S-[(15)N]Nitrosoglutathione (GS(15)NO) served as internal standard. Fresh blood was treated with NEM/serine/borate/EDTA, plasma spiked with GS(15)NO (50nM) was ultrafiltered (cut-off 10kDa) and 10µL aliquots of the ultrafiltrate were analyzed by UPLC-MS/MS. Five HILIC columns and an Acquity UPLC BH amide column were tested. The mobile phase was acetonitrile-water (70:30, v/v), contained 20mM ammonium formate, had a pH value of 7, and was pumped isocratically (0.5mL/min). The Nucleoshell column allowed better LC performance and higher MS sensitivity. The retention time of GSNO was about 1.1min. Quantification was performed by selected-reaction monitoring the mass transition m/z 337 ([M+H](+))→m/z 307 ([M+H(14)NO](+)) for GSNO (i.e., GS(14)NO) and m/z 338 ([M+H](+))→m/z 307 ([M+H(15)NO](+)) for GS(15)NO. NEM/serine/borate/EDTA was found to stabilize GSNO in human plasma. The method was validated in human plasma (range, 0-300nM) using 50nM GS(15)NO. Accuracy and precision were in generally acceptable ranges. A considerable matrix effect was observed, which was however outweighed by the internal standard GS(15)NO. In freshly prepared plasma from heparinized blood donated by 10 healthy subjects, no endogenous GSNO was determined above 2.8nM, the limit of quantitation (LOQ) of the method. This study challenges previously reported GSNO plasma concentrations being far above the present method LOQ value and predicts that the concentration of low-molecular-mass and high-molecular-mass S-nitrosothiols are in the upper pM- and lower nM-range, respectively.

Microevolution of the major common Pseudomonas aeruginosa clones C and PA14 in cystic fibrosis lungs.

Clones C and PA14 are the worldwide most abundant clonal complexes in the Pseudomonas aeruginosa population. The microevolution of clones C and PA14 was investigated in serial cystic fibrosis (CF) airway isolates collected over 20 years since the onset of colonization. Intraclonal evolution in CF lungs was resolved by genome sequencing of first, intermediate and late isolates and subsequent multimarker SNP genotyping of the whole strain panel. Mapping of sequence reads onto the P. aeruginosa PA14 reference genome unravelled an intraclonal and interclonal sequence diversity of 0.0035% and 0.68% respectively. Clone PA14 diversified into three branches in the patient's lungs, and the PA14 population acquired 15 nucleotide substitutions and a large deletion during the observation period. The clone C genome remained invariant during the first 3 years in CF lungs; however, 15 years later 947 transitions and 12 transversions were detected in a clone C mutL mutant strain. Key mutations occurred in retS, RNA polymerase, multidrug transporter, virulence and denitrification genes. Late clone C and PA14 persistors in the CF lungs were compromised in growth and cytotoxicity, but their mutation frequency was normal even in mutL mutant clades.

Inertance measurements by jet pulses in ventilated small lungs after perfluorochemical liquid (PFC) applications.

Perfluorochemical liquid (PFC) liquids or aerosols are used for assisted ventilation, drug delivery, lung cancer hyperthermia and pulmonary imaging. The aim of this study was to investigate the effect of PFC liquid on the inertance (I) of the respiratory system in newborn piglets using partial liquid ventilation (PLV) with different volumes of liquid. End-inspiratory (I(in)) and end-expiratory (I(ex)) inertance were measured in 15 ventilated newborn piglets (age < 12 h, mean weight 724 +/- 93 g) by brief flow pulses before and 80 min after PLV using a PFC volume (PF5080, 3 M) of 10 ml kg(-1) (N = 5) or 30 ml kg(-1) (N = 10). I was calculated from the imaginary part of the measured respiratory input impedance by regression analysis. Straight tubes with 2-4 mm inner diameter were used to validate the equipment in vitro by comparison with the analytically calculated values. In vitro measurements showed that the measuring error of I was <5% and that the reproducibility was better than 1.5%. The correlation coefficient of the regression model to determine I was >0.988 in all piglets. During gas ventilation, I(in) and I(ex) (mean +/- SD) were 31.7 +/- 0.8 Pa l(-1) s(2) and 33.3 +/- 2.1 Pa l(-1) s(2) in the 10 ml group and 32.4 +/- 0.8 Pa l(-1) s(2) and 34.0 +/- 2.5 Pa l(-1) s(2) in the 30 ml group. However, I of the 3 mm endotracheal tube (ETT) used was already 26.4 Pa l(-1) s(2) (about 80% of measured I). During PLV, there was a minimal increase of I(in) to 33.1 +/- 2.5 Pa l(-1) s(2) in the 10 ml group and to 34.5 +/- 2.7 Pa l(-1) s(2) in the 30 ml group. In contrast, the increase of I(ex) was dramatically larger (p < 0.001) to 67.7 +/- 13.3 Pa l(-1) s(2) and to 74.8 +/- 9.3 Pa l(-1) s(2) in the 10 ml and 30 ml groups, respectively. Measurements of I by jet pulses in intubated small animals are reproducible. PFC increases the respiratory inertance, but the magnitude depends considerably on its spatial distribution which changes during the breathing cycle. Large differences between I(in) and I(ex) are an indicator for liquid in airways or the ETT.

Measurement of changes in respiratory mechanics during partial liquid ventilation using jet pulses.

OBJECTIVE: To compare the changes in respiratory mechanics within the breathing cycle in healthy lungs between gas ventilation and partial liquid ventilation using a special forced-oscillation technique. DESIGN: Prospective animal trial. SETTINGS: Animal laboratory in a university setting. SUBJECTS: A total of 12 newborn piglets (age, <12 hrs; mean weight, 725 g). INTERVENTIONS: After intubation and instrumentation, lung mechanics of the anesthetized piglets were measured by forced-oscillation technique at the end of inspiration and the end of expiration. The measurements were performed during gas ventilation and 80 mins after instillation of 30 mL/kg perfluorocarbon PF 5080. MEASUREMENTS AND MAIN RESULTS: Brief flow pulses (width, 10 msec; peak flow, 16 L/min) were generated by a jet generator to measure the end-inspiratory and the end-expiratory respiratory input impedance in the frequency range of 4-32 Hz. The mechanical variables resistance, inertance, and compliance were determined by model fitting, using the method of least squares. At least in the lower frequency range, respiratory mechanics could be described adequately by an RIC single-compartment model in all piglets. During gas ventilation, the respiratory variables resistance and inertance did not differ significantly between end-inspiratory and end-expiratory measurements (mean [sd]: 4.2 [0.7] vs. 4.1 [0.6] kPa x L(-1) x sec, 30.0 [3.2] vs. 30.7 [3.1] Pa x L(-1) x sec2, respectively), whereas compliance decreased during inspiration from 14.8 (2.0) to 10.2 (2.4) mL x kPa(-1) x kg(-1) due to a slight lung overdistension. During partial liquid ventilation, the end-inspiratory respiratory mechanics was not different from the end-inspiratory respiratory mechanics measured during gas ventilation. However, in contrast to gas ventilation during partial liquid ventilation, compliance rose from 8.2 (1.0) to 13.0 (3.0) mL x kPa(-1) x kg(-1) during inspiration. During expiration, when perfluorocarbon came into the upper airways, both resistance and inertance increased considerably (mean with 95% confidence interval) by 34.3% (23.1%-45.8%) and 104.1% (96.0%-112.1%), respectively. CONCLUSIONS: The changes in the respiratory mechanics within the breathing cycle are considerably higher during partial liquid ventilation compared with gas ventilation. This dependence of lung mechanics from the pulmonary gas volume hampers the comparability of dynamic measurements during partial liquid ventilation, and the magnitude of these changes cannot be detected by conventional respiratory-mechanical analysis using time-averaged variables.