"Exotic" seeds from Southern Africa as potential Novel Foods? - Chemical composition of manketti nuts (Schinziophyton rautanenii) and ushivi beans (Guibourtia coleosperma).

Various species of Southern African plants and their edible seeds have gained huge importance due to positive health aspects, and there is increasing interest to introduce such seeds as Novel Food on the international market. Especially the seeds of Schinziophyton rautanenii (manketti) and Guibourtia coleosperma (ushivi) could have great potential as a food and food ingredient. Hence, extensive analyses on the chemical composition of manketti nuts and ushivi beans including the analysis of total solids, protein and fat content, soluble carbohydrates, ash, total and free amino acids, biogenic amines and polyamines, fatty acid profile as well as the content of certain B-vitamins and tocopherols were performed. Results obtained showed a valuable nutritional composition, e.g., a true protein content of 22.6% with a ratio of essential amino acids to total amino acid composition of 48% in manketti nuts, while ushivi beans had a true protein content of 8.2% with a similar ratio of essential to total amino acids (45%). Lipid content was 54.1% in manketti nuts, ushivi beans had a value of 7.7%. In both, linoleic acid was the most abundant. Furthermore, ushivi beans had high amounts of vitamin B1 and B2.

Geospatial Information Research: State of the Art, Case Studies and Future Perspectives.

Geospatial information science (GI science) is concerned with the development and application of geodetic and information science methods for modeling, acquiring, sharing, managing, exploring, analyzing, synthesizing, visualizing, and evaluating data on spatio-temporal phenomena related to the Earth. As an interdisciplinary scientific discipline, it focuses on developing and adapting information technologies to understand processes on the Earth and human-place interactions, to detect and predict trends and patterns in the observed data, and to support decision making. The authors - members of DGK, the Geoinformatics division, as part of the Committee on Geodesy of the Bavarian Academy of Sciences and Humanities, representing geodetic research and university teaching in Germany - have prepared this paper as a means to point out future research questions and directions in geospatial information science. For the different facets of geospatial information science, the state of art is presented and underlined with mostly own case studies. The paper thus illustrates which contributions the German GI community makes and which research perspectives arise in geospatial information science. The paper further demonstrates that GI science, with its expertise in data acquisition and interpretation, information modeling and management, integration, decision support, visualization, and dissemination, can help solve many of the grand challenges facing society today and in the future.

"A2 milk" authentication using isoelectric focusing and different PCR techniques.

Genetic variants of milk proteins have attracted great interest for decades as they are related to important issues such as the composition and technological properties of milk. More recently, an "A1/A2 hypothesis" was developed saying that ß-casein variant A1 may be a dietary risk factor for cardiovascular diseases, type 1 diabetes, sudden infant death syndrome and neurological disorders due to the release of ß-casomorphin-7, whereas no evidence for such adverse effects was assumed for ß-casein A2. Thus, the aim of this study was to adapt and establish analytical methods for the identification of genetic variants of ß-casein using isoelectric focusing of milk proteins as well as appropriate PCR techniques. Allele-specific polymerase chain reaction (AS-PCR) proved to be a reliable method for differentiating most common ß-casein variants (A1, A2, B, C), amplification-created restriction site (ACRS)-PCR using three different restriction enzymes allowed also the detection of variant A3, and the restriction fragment length polymorphism (RFLP)-PCR method enabled the reliable discrimination between A2 (homozygote/heterozygote) and non-A2 animals. Since traces of ß-casein A1 were also found in commercial "A2 milk" in Austria, the authentication of such expensive dairy products is urgently recommended, both by genotyping of all dairy cows at farm level (to confirm that all cows are homozygous ß-casein A2A2) and by screening commercial products on the market (to confirm the absence of ß-casein variants A1, B, and C in dairy products labelled "A2 milk") to protect consumers from this unexpected fraud.

Development and validation of a real-time PCR assay to detect Cannabis sativa in food.

Regarding the prospective investigation of food authenticity and adulteration the aim of the present study was the development and validation of a real-time PCR assay to identify hemp (Cannabis sativa) which has gained increasing importance as a valuable food ingredient. The assay targets a specific spacer DNA sequence in Cannabis sativa chloroplasts and detects 1.5 pg hemp DNA, which is equivalent to 18 copies/µL. Corresponding to the very low LOD (0.00031 ng/µL) the method allows the detection of hemp even in the infinitesimal concentration of contaminants. Due to a SNP in position 603, hemp can be identified unequivocally and discriminated from its closest relative hops (Humulus lupulus). The PCR method shows no cross-reactivity with 39 of 46 tested plant species. Low cross-reactivity with mulberry, stinging nettle, lavender, cornflower, wine, figs and hops can be neglected, because the Δ Ct-values are > 14, and the obtained Ct-values are beyond the cut-off for a positive assessment (Ct-values ≤ 33). Moreover, the suitability of the method to identify hemp as a food ingredient was proved by analysing diverse food products such as chocolate or cookies.

A novel UHPLC method for determining the degree of coffee roasting by analysis of furans.

The aim of this study was to develop a multi-analyte UHPLC method for furans and to apply it to commercial coffee samples as well as commercial roasting trials. Furans, as rising time-temperature indicators (TTIs), promised to be an alternative to unsatisfactory roasting temperature measurements. Consequently, a UHPLC-UV method for the determination of 5-hydroxymethyl-2-furfural (HMF), 5-hydroxymethyl-2-furoic acid (HMFA), 2-furfural (F), 5-methylfurfural (MF), 2-furyl methyl ketone (FMC), 2-furoic acid (FA), and for 3-caffeoylquinic acid (3-CQA) was developed and validated. Commercial roasted coffee beans contained 77.7-322 mg/kg HMF, 73.3-158 mg/kg HMFA, 109-200 mg/kg 2-F, 157-209 mg/kg MF, 12.3-32.8 mg/kg FMC, and 137-205 mg/kg FA. Roasting trial samples showed strong rising HMF contents (max.: Arabica: 769 mg/kg, Robusta: 364 mg/kg) followed by a distinct decline. Only MF and FA appeared as steady rising TTIs in the roasting process in Arabica and Robusta beans. 3-CQA fitted well as a decreasing TTI as expected.

High-throughput quantitation of bovine milk proteins and discrimination of commercial milk types by external cavity-quantum cascade laser spectroscopy and chemometrics.

Analysis of bovine milk proteins is crucial in many food and non-food industrial applications, nevertheless labour-intensive wet-chemical, low-throughput methods are still routinely used. In this work, external cavity-quantum cascade laser (EC-QCL) mid-infrared spectroscopy is employed as a rapid method for protein analysis of commercial bovine milk. Combined analysis of the amide I and II bands enabled quantitation of individual proteins (casein, ß-lactoglobulin, α-lactalbumin) and total protein content. IR spectra of spiked and diluted milk samples were employed for calibration of the target analytes in the presence of a complex matrix by partial least squares (PLS) regression modelling. A sample set of different milk types (pasteurized; differently processed extended shelf life, ESL; ultra-high temperature, UHT) was analysed, and results agreed well with reference methods. Quantitation of temperature sensitive proteins enables detailed distinction between milk types experiencing different heat loads during processing, and discrimination between diverse bovine milk types is successfully demonstrated.

Determination of vitamin B12 in four edible insect species by immunoaffinity and ultra-high performance liquid chromatography.

Insects are rich in major nutrients, such as protein and fat. Recently, minor nutrients like vitamins have become the subjects of interest in insects. Hence, this study reports on the development and validation of a method for the determination of vitamin B12 in mealworm (Tenebrio molitor larvae), cricket (Gryllus assimilis), grasshopper (Locusta migratoria) and cockroach (Shelfordella lateralis), using an ultra-high performance liquid chromatography approach with preliminary immunoaffinity chromatography sample preparation. The method was validated regarding linearity, specificity, accuracy and precision, as well as limits of detection/quantification, and was found to be satisfactory for the desired application. Found levels of vitamin B12 were 1.08 µg/100 g for mealworm, 2.88 µg/100 g for cricket, 0.84 µg/100 g for grasshopper, and 13.2 µg/100 g dry weight for cockroach, representing the first validated report on the content of vitamin B12 in edible insects. Observed interferences are likely caused by the presence of pseudovitamin B12.

To what extent does the air flow initialisation of the ENVI-met model affect human heat stress simulated in a common street canyon?

This study concerns the effects of the air flow initialisation of the ENVI-met model on simulated human heat stress in a fictive E-W street canyon with different aspect ratios that is typical of Central European cities. Human heat stress is described by near-surface air temperature (Ta), mean radiant temperature (Tmrt) and physiologically equivalent temperature (PET). The numerical simulations, which are performed for a Central European heat wave day in order to consider the increase of severe heat due to regional climate change, are based on the version 4.0 BETA of ENVI-met. The simulation results in terms of grid-related Ta, Tmrt and PET values as well as mean values for both sidewalks of the street canyon are averaged over the period 10-16 CET, because they should be representative of outdoor human heat stress in Central European cities. The simulation results point to the significance of the type of inflow direction in relation to the orientation of the street canyon, i.e. whether it is flowed parallel or across. The type of inflow direction determines the modification of the inflow speed within the street canyon. Due to its physical basis, mean Tmrt does not show a noticeable impact by the inflow conditions. They also influence mean Ta relatively low. However, PET is much more affected by the inflow conditions as it depends on the local wind speed. This impact can reach the magnitude of thermal grassland effects within urban quarters, i.e. it cannot be ignored under a human-biometeorological perspective.

Thermal comfort of pedestrians in an urban street canyon is affected by increasing albedo of building walls.

Numerical simulations based on the ENVI-met model were carried out for an E-W street canyon in the city of Stuttgart (Southwest Germany) to analyse the effect of increased albedo of building walls on outdoor human thermal comfort. It was quantified by air temperature (Ta), mean radiant temperature (Tmrt) and physiologically equivalent temperature (PET). The simulations were conducted on 4 August 2003 as a heat wave day that represents a typical scenario for future summer weather in Central Europe. The simulation results presented for 13 CET and averaged over the period 10-16 CET are focused on pedestrians on both sidewalks. For the initial situation, i.e. albedo of 0.2, human heat stress indicated by mean PET is by 26% lower on the N-facing than on the S-facing sidewalk, while this reduction amounts to 42% for mean Tmrt. Mean Ta does not show any spatial differentiation. The systematic albedo increment by 0.2 from 0.2 to 0.8 leads to a linear increase of outdoor human heat stress in terms of Tmrt and PET. For both variables, this increase is more pronounced on the N-facing than on the S-facing sidewalk. Mean relative Ta shows the tendency of a minimal increase with rising albedo. The results were achieved for the usual standardised human-biometeorological reference person. Its substitution by two other types of male and female pedestrians, respectively, which are statistically characteristic of human conditions in Germany, does not reveal any significant change in the results.

Fast quantification of bovine milk proteins employing external cavity-quantum cascade laser spectroscopy.

Analysis of proteins in bovine milk is usually tackled by time-consuming analytical approaches involving wet-chemical, multi-step sample clean-up procedures. The use of external cavity-quantum cascade laser (EC-QCL) based IR spectroscopy was evaluated as an alternative screening tool for direct and simultaneous quantification of individual proteins (i.e. casein and ß-lactoglobulin) and total protein content in commercial bovine milk samples. Mid-IR spectra of protein standard mixtures were used for building partial least squares (PLS) regression models. A sample set comprising different milk types (pasteurized; differently processed extended shelf life, ESL; ultra-high temperature, UHT) was analysed and results were compared to reference methods. Concentration values of the QCL-IR spectroscopy approach obtained within several minutes are in good agreement with reference methods involving multiple sample preparation steps. The potential application as a fast screening method for estimating the heat load applied to liquid milk is demonstrated.

Sleep in patients with disorders of consciousness characterized by means of machine learning.

Sleep has been proposed to indicate preserved residual brain functioning in patients suffering from disorders of consciousness (DOC) after awakening from coma. However, a reliable characterization of sleep patterns in this clinical population continues to be challenging given severely altered brain oscillations, frequent and extended artifacts in clinical recordings and the absence of established staging criteria. In the present study, we try to address these issues and investigate the usefulness of a multivariate machine learning technique based on permutation entropy, a complexity measure. Specifically, we used long-term polysomnography (PSG), along with video recordings in day and night periods in a sample of 23 DOC; 12 patients were diagnosed as Unresponsive Wakefulness Syndrome (UWS) and 11 were diagnosed as Minimally Conscious State (MCS). Eight hour PSG recordings of healthy sleepers (N = 26) were additionally used for training and setting parameters of supervised and unsupervised model, respectively. In DOC, the supervised classification (wake, N1, N2, N3 or REM) was validated using simultaneous videos which identified periods with prolonged eye opening or eye closure.The supervised classification revealed that out of the 23 subjects, 11 patients (5 MCS and 6 UWS) yielded highly accurate classification with an average F1-score of 0.87 representing high overlap between the classifier predicting sleep (i.e. one of the 4 sleep stages) and closed eyes. Furthermore, the unsupervised approach revealed a more complex pattern of sleep-wake stages during the night period in the MCS group, as evidenced by the presence of several distinct clusters. In contrast, in UWS patients no such clustering was found. Altogether, we present a novel data-driven method, based on machine learning that can be used to gain new and unambiguous insights into sleep organization and residual brain functioning of patients with DOC.

Determination of the native forms of vitamin B1 in bovine milk using a fast and simplified UHPLC method.

The aim of this study was to develop a high-throughput UHPLC method for the determination vitamin B1 active compounds; thiamin, thiamin monophosphate and thiamin diphosphate in bovine milk. In order to sustain the native vitamin B1 phosphorus esters, sample preparation is crucial. Various acids as well as commonly used enzymes and their enzyme mixtures were compared. Method accuracy was confirmed using certified reference material as well as comparison with the corresponding CEN method, and was found to be satisfactory. Studied milk samples showed significant amounts of thiamin monophosphate, which can make up to 53.9% of the total vitamin B1 content in commercial milk, and up to 78% in raw milk. Moreover, a tremendous variation of the total content of vitamin B1 was observed between single cows, which ranged from 0.24mg/L up to 0.54mg/L of total vitamin B1.

External cavity-quantum cascade laser (EC-QCL) spectroscopy for protein analysis in bovine milk.

The analytical determination of bovine milk proteins is important in food and non-food industrial applications and yet, rather labour-intensive wet-chemical, low-throughput methods have been employed since decades. This work proposes the use of external cavity-quantum cascade laser (EC-QCL) spectroscopy for the simultaneous quantification of the most abundant bovine milk proteins and the total protein content based on the chemical information contained in mid-infrared (IR) spectral features of the amide I band. Mid-IR spectra of protein standard mixtures were used for building partial least squares (PLS) regression models. Protein concentrations in commercial bovine milk samples were calculated after chemometric compensation of the matrix contribution employing science-based calibration (SBC) without sample pre-processing. The use of EC-QCL spectroscopy together with advanced multivariate data analysis allowed the determination of casein, α-lactalbumin, ß-lactoglobulin and total protein content within several minutes.

Analytical assessment of the intense heat load of whipping cream, coffee cream, and condensed milk at retail in Austria and Germany.

Time temperature integrators (TTIs) are useful tools in estimating the heat load applied on differently processed dairy products. The objective of this study was to analyze and assess three TTIs - lactulose, furosine, and acid-soluble ß-lactoglobulin (ß-Lg) - in 70 high heated dairy products at retail in Austria and Germany comprising whipping cream, coffee cream/milk, and condensed milk products. While ß-Lg was not appropriate to evaluate the heat load of these products, furosine and especially lactulose increased with rising intensity of heat treatment, and are appropriate to distinguish between several heating categories analyzed. Pasteurized (n = 8) and "heat treated" (n = 5) whipping cream samples showed lowest furosine (48 ± 14/ 45 ± 19 mg.100 g-1 protein) and low lactulose (29 ± 10/57 ± 28 mg.L-1) concentrations, followed by ESL whipping cream (n = 10), ESL coffee cream (n = 1), and UHT whipping cream (n = 10) (furosine = 72 ± 37/71/161 ± 30 mg.100 g-1 protein; lactulose = 56 ± 41/161/195 ± 39 mg.L-1), respectively. Sterilized condensed milk samples (n = 14) showed the highest concentrations of both TTIs and could be clearly separated from UHT treated samples (n = 5) (furosine = 491 ± 196/216 ± 46 mg.100 g-1 protein; lactulose = 1997 ± 658/409 ± 161 mg.L-1), whereas the so-called heat-treated samples (n = 9) had a heat load in between showing an extreme range of variation for both TTIs.

Validation of the mean radiant temperature simulated by the RayMan software in urban environments.

The RayMan software is worldwide applied in investigations on different issues in human-biometeorology. However, only the simulated mean radiant temperature (T mrt) has been validated so far in a few case studies. They are based on T mrt values, which were experimentally determined in urban environments by use of a globe thermometer or applying the six-directional method. This study analyses previous T mrt validations in a comparative manner. Their results are extended by a recent validation of T mrt in an urban micro-environment in Freiburg (southwest Germany), which can be regarded as relatively heterogeneous due to different shading intensities by tree crowns. In addition, a validation of the physiologically equivalent temperature (PET) simulated by RayMan is conducted for the first time. The validations are based on experimentally determined T mrt and PET values, which were calculated from measured meteorological variables in the daytime of a clear-sky summer day. In total, the validation results show that RayMan is capable of simulating T mrt satisfactorily under relatively homogeneous site conditions. However, the inaccuracy of simulated T mrt is increasing with lower sun elevation and growing heterogeneity of the simulation site. As T mrt represents the meteorological variable that mostly governs PET in the daytime of clear-sky summer days, the accuracy of simulated T mrt is mainly responsible for the accuracy of simulated PET. The T mrt validations result in some recommendations, which concern an update of physical principles applied in the RayMan software to simulate the short- and long-wave radiant flux densities, especially from vertical building walls and tree crowns.

A novel ultra-high performance liquid chromatography method for the rapid determination of ß-lactoglobulin as heat load indicator in commercial milk samples.

The level of undenatured acid-soluble ß-lactoglobulin can be used as an indicator to assess the heat load applied to liquid milk, thus further allowing the discrimination between milk originating from different thermal production processes. In this work, a new UHPLC method for the rapid determination of bovine ß-lactoglobulin in 1.8min only (total runtime 3min) is presented using simple UV detection at 205nm. Separation selectivity for possibly co-eluting other major whey proteins (bovine serum albumin, lactoferrin, α-lactalbumin, immunoglobulin G) was verified, and the method validated for the analysis of liquid milk samples regarding linearity (20-560µg/mL, R(2)>0.99), instrumentation precision (RSDs<2.8%), limits of detection and quantification (7 and 23mg/L milk), repeatability of sample work-up (RSDs≤2.6%) and method recovery (103%). In total, 71 commercial liquid milk samples produced using different preservation techniques (e.g., thermal or mechanical treatment), hence featuring different applied heat loads, were profiled for their intrinsic undenatured acid-soluble ß-lactoglobulin levels. As expected, pasteurized milk showed the highest concentrations clearly above 3000mg/L due to pasteurization being the mildest thermal treatment, while in contrast, ultra-high temperature heated milk featured the lowest amounts (<200mg/L). For extended shelf life (ESL) milk, quite diverse levels were determined ranging from ∼100 up to 4000mg/L, thus clearly illustrating variable applied heat loads and impacts on the "nativeness" of milk essentially due to the fact that the production technologies used for ESL milk may differ significantly, and are currently not regulated in the EU.

Application of UHPLC for the determination of free amino acids in different cheese varieties.

A rapid ultra-high performance liquid chromatography (UHPLC) protocol for the determination of amino acids as their respective 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) derivatives was successfully applied for assessing free amino acid levels in commercial cheese samples representing typical product groups (ripening protocols) in cheesemaking. Based on the Waters AccQ.Tag™ method as a high performance liquid chromatography (HPLC) amino acid solution designed for hydrolyzate analyses, method adaptation onto UHPLC was performed, and detection of AQC derivatives was changed from former fluorescence (λ(Ex) 250 nm/λ(Em) 395 nm) to UV (254 nm). Compared to the original HPLC method, UHPLC proved to be superior by facilitating excellent separations of 18 amino acids within 12 min only, thus demonstrating significantly shortened runtimes (>35 min for HPLC) while retaining the original separation chemistry and amino acid elution pattern. Free amino acid levels of the analyzed cheese samples showed a high extent of variability depending on the cheese type, with highest total amounts found for original Italian extra-hard cheeses (up to 9,000 mg/100 g) and lowest for surface mold- or bacterial smear-ripened soft cheeses (200-600 mg/100 g). Despite the intrinsic variability in both total and specific concentrations, the established UHPLC method enabled reliable and interference-free amino acid profiling throughout all cheese types, thus demonstrating a valuable tool to generate high quality data for the characterization of cheese ripening.

Application of UHPLC for the simultaneous analysis of free amino acids and biogenic amines in ripened acid-curd cheeses.

A new UHPLC method for the simultaneous determination of amino acids and biogenic amines in a single run, and its first application to profile ripened acid-curd cheeses was presented. After pre-column derivatization with 6-aminoquinolyl-N-hydroxy succinimidyl carbamate (AQC), 23 amino acids and 15 amines were separated in 9min only (12min total run time), and eluates monitored using their UV response at 249nm. Limits of detection (0.05-0.29mg/100g) and quantification (0.16-0.97mg/100g), repeatability for sample preparation (1.0-6.1% RSD) and method recoveries (83-120%) were found suitable for cheese analysis. In total, 47 acid-curd cheeses classified into sub-groups like cooked, Quargel-type or grey cheeses were analyzed for their free amino acid and amine (histamine, tyramine, putrescine, cadaverine, and tryptamine) contents, which (as expected) were highlighted by a great variability. Total free amino acid levels ranged between less than 100 and more than 4000mg/100g (median 567mg/100g), implying that for some cheeses less or not ripened/fresh quark was used for production or, in contrast, a higher degree of proteolysis had occurred. For the sum of biogenic amines, median concentration was determined at 7.0mg/100g, while only 5% of all cheeses had levels higher than 161.9mg/100g. Thus, the obtained results suggest quite acceptable biogenic amine levels for (mostly underrated) ripened acid-curd cheeses, although partly exceptional high concentrations (>250mg/100g) were indeed observed in individual samples.

Analysis and simulation of dynamic response behavior of Scots pine trees to wind loading.

This paper presents an empirical approach for the decomposition, simulation, and reconstruction of wind-induced stem displacement of plantation-grown Scots pine trees. Results from singular spectrum analysis (SSA) allow a low-dimensional characterization of the complex and complicated tree motion patterns in response to non-destructive wind excitation. Since motion of the sample trees was dominated by sway in the first mode, the application of SSA on time series of sample trees' stem displacement yielded characteristic and distinguishable non-oscillatory trend components, quasi-oscillatory sway, and noise, of which only the non-oscillatory components were correlated directly with wind characteristics. Although sway in the range of the dominant damped fundamental frequency dominated the measured stem displacement signals, it was almost decoupled from near-surface airflow. The ability to discriminate SSA-components is demonstrated based on correlation and spectral analysis. These SSA-components, as well as wind speed measured in the canopy space of the Scots pine forest, were used to train neural networks, which could then reasonably simulate tree response to wind excitation.

Quantification of cow's milk percentage in dairy products - a myth?

The substitution of ewe's and goat's milk for cheaper cow's milk is still a fraudulent practice in the dairy industry. Moreover, soy-based products (e.g., soy milk, yoghurt) have to be checked for cow's milk as they are an alternative for people suffering from an allergy against bovine milk proteins. This work reports the evaluation of different protein-based electrophoretic methods and DNA-based techniques for the qualitative detection as well as the quantitative determination of cow's milk percentage in dairy and soy milk products. Isoelectric focusing (IEF) of γ-caseins using an optimized pH gradient was appropriate not only for the detection of cow's milk, but also for an estimation of cow's milk percentage in mixed-milk cheese varieties. Urea-polyacrylamide gel electrophoresis (PAGE) proved the method of choice to detect cow's milk in soy milk products, whereas IEF and SDS-PAGE of proteins were not applicable due to false-positive results. Polymerase chain reaction (PCR) analysis was used to confirm the results of protein-based electrophoretic methods. Problems inherent in quantitative analysis of cow's milk percentage using protein-based techniques and even more using DNA-based methods were emphasized. Applicability of quantitative real-time PCR for the determination of cow's milk percentage in mixed-milk cheese was shown to be hampered by several factors (e.g., somatic cell count of milk; technological parameters influencing the final DNA concentration in ripened commercial cheese samples). The implementation of certified reference standards (of major relevant cheese groups) containing 50% cow's milk was urgently recommended to enable at least a yes/no decision in commercial mixed-milk cheese samples.