Soil self-cleaning capacity: Removal of organic compounds during sub-surface irrigation with sewage effluent.

Globally, the reuse of treated sewage effluent for irrigation purposes is increasingly encouraged as a practical solution against the mismatch between the demand for and availability of freshwater resources. The reuse of sewage effluent for sub-surface irrigation (SSI) in agriculture serves the dual purpose of supplying water to crops and diminishing emissions of contaminants of emerging concern (CoECs) into surface water. To investigate such reuse, in a real scale cropland with SSI using sewage effluent, from September 2017 to March 2019 including the extremely dry year 2018, residues were followed of 133 CoECs as related to their physicochemical properties and quantified by liquid chromatography coupled to high-resolution mass spectrometry. Of the 133 target CoECs, 89 were retrieved in the field, most non-detect CoECs have low persistency. During the growing season with sub-surface irrigation, CoECs spread to the shallow groundwater and rhizosphere. Significantly lower concentrations are found between infiltration pipes as compared to directly next to the pipes in shallow groundwater for all persistency-mobility classes. CoECs belonging to the class pm (low persistency and low mobility) or class PM (high persistency and high mobility) class show no change amongst their removal in the rhizosphere and groundwater in a dry versus normal year. CoECs belonging to the class pM (low persistency and high mobility) show high seasonal dynamics in the rhizosphere and shallow groundwater, indicating that these CoECs break down. CoECs of the class Pm (high persistency and low mobility) only significantly build up in the rhizosphere next to infiltration pipes. Climatic conditions with dry summers and precipitation surplus and drainage in winter strongly affect the fate of CoECs. During the dry summer of 2018 infiltrated effluent is hardly diluted, resulting in significantly higher concentrations for the CoECs belonging to the classes pM and Pm. After the extremely dry year of 2018, cumulative concentrations are still significantly higher, while after a normal year during winter precipitation surplus removes CoECs. For all persistency-mobility classes in the shallow groundwater between the pipes, we find significant removal efficiencies. For the rhizosphere between the pipes, we find the same except for Pm. Next to the pipes however we find no significant removal for all classes in both the rhizosphere and shallow groundwater and even significant accumulation for Pm. For this group of persistent moderately hydrophobic CoECs risk characterization ratio's were calculated for the period of time with the highest normalized concentration. None of the single-chemical RCRs are above one and the ΣRCR is also far below one, implying sufficiently safe ambient exposures. Overall the deeper groundwater (7.0-11.8 m below soil surface) has the lowest response to the sub-surface irrigation for all persistency-mobility. When adopting a SSI STP effluent reuse system care must be taken to monitor the CoECs that are (moderately) hydrophobic as these can build up in the SSI system. For the deeper groundwater and for the discharge to the surface water, we find significant removal for the pM and the PM class but not for other classes. In conclusion, relatively high removal efficiencies are shown benefiting the surface waters that would otherwise receive the STP effluent directly.

Inflammatory and regenerative processes in bioresorbable synthetic pulmonary valves up to two years in sheep-Spatiotemporal insights augmented by Raman microspectroscopy.

In situ heart valve tissue engineering is an emerging approach in which resorbable, off-the-shelf available scaffolds are used to induce endogenous heart valve restoration. Such scaffolds are designed to recruit endogenous cells in vivo, which subsequently resorb polymer and produce and remodel new valvular tissue in situ. Recently, preclinical studies using electrospun supramolecular elastomeric valvular grafts have shown that this approach enables in situ regeneration of pulmonary valves with long-term functionality in vivo. However, the evolution and mechanisms of inflammation, polymer absorption and tissue regeneration are largely unknown, and adverse valve remodeling and intra- and inter-valvular variability have been reported. Therefore, the goal of the present study was to gain a mechanistic understanding of the in vivo regenerative processes by combining routine histology and immunohistochemistry, using a comprehensive sheep-specific antibody panel, with Raman microspectroscopy for the spatiotemporal analysis of in situ tissue-engineered pulmonary valves with follow-up to 24 months from a previous preclinical study in sheep. The analyses revealed a strong spatial heterogeneity in the influx of inflammatory cells, graft resorption, and foreign body giant cells. Collagen maturation occurred predominantly between 6 and 12 months after implantation, which was accompanied by a progressive switch to a more quiescent phenotype of infiltrating cells with properties of valvular interstitial cells. Variability among specimens in the extent of tissue remodeling was observed for follow-up times after 6 months. Taken together, these findings advance the understanding of key events and mechanisms in material-driven in situ heart valve tissue engineering. STATEMENT OF SIGNIFICANCE: This study describes for the first time the long-term in vivo inflammatory and regenerative processes that underly in situ heart valve tissue engineering using resorbable synthetic scaffolds. Using a unique combinatorial analysis of immunohistochemistry and Raman microspectroscopy, important spatiotemporal variability in graft resorption and tissue formation was pinpointed in in situ tissue-engineered heart valves, with a follow-up time of up to 24 months in sheep. This variability was correlated to heterogenous regional cellular repopulation, most likely instigated by region-specific differences in surrounding tissue and hemodynamics. The findings of this research contribute to the mechanistic understanding of in situ tissue engineering using resorbable synthetics, which is necessary to enable rational design of improved grafts, and ensure safe and robust clinical translation.

A systems approach to understand microplastic occurrence and variability in Dutch riverine surface waters.

Assessment methods on data quality and environmental variability are lacking for microplastics (MP). Here we assess occurrence and variability of MP number concentrations in two Dutch rivers. Strict QA/QC procedures were applied to identify MP using Fourier-transform infrared (FTIR) microscopy followed by state of the art automated image analysis. For a series of randomly selected, yet ever smaller subareas of filters, we assessed how accurately MP numbers and polymer types are represented during partial filter analysis. Levels of uncertainty were acceptable when analysing 50% of a filter during chemical mapping, and when identifying at least a subset of 50 individual particles with attenuated total reflection (ATR)-FTIR. Applying these guidelines, MP number concentrations between 67 and 11532 MP m-3 were detected in Dutch riverine surface waters. Spatial differences caused MP number concentrations to vary by two orders of magnitude. Temporal differences were lower and induced a maximum variation of one order of magnitude. In total, 26 polymer types were identified, the most common were polyethylene (23%), polypropylene (19.7%) and ethylene propylene diene monomer rubber (18.3%). The highest diversity of polymer types was found for small MPs, whereas MP larger than 1 mm was scarce and almost exclusively made of polyethylene or polypropylene. Virtually all sampling locations revealed MP number concentrations that are considerably below known effect thresholds for anticipated adverse ecological effects.

Modular operation of microfluidic chips for highly parallelized cell culture and liquid dosing via a fluidic circuit board.

Microfluidic systems enable automated and highly parallelized cell culture with low volumes and defined liquid dosing. To achieve this, systems typically integrate all functions into a single, monolithic device as a "one size fits all" solution. However, this approach limits the end users' (re)design flexibility and complicates the addition of new functions to the system. To address this challenge, we propose and demonstrate a modular and standardized plug-and-play fluidic circuit board (FCB) for operating microfluidic building blocks (MFBBs), whereby both the FCB and the MFBBs contain integrated valves. A single FCB can parallelize up to three MFBBs of the same design or operate MFBBs with entirely different architectures. The operation of the MFBBs through the FCB is fully automated and does not incur the cost of an extra external footprint. We use this modular platform to control three microfluidic large-scale integration (mLSI) MFBBs, each of which features 64 microchambers suitable for cell culturing with high spatiotemporal control. We show as a proof of principle that we can culture human umbilical vein endothelial cells (HUVECs) for multiple days in the chambers of this MFBB. Moreover, we also use the same FCB to control an MFBB for liquid dosing with a high dynamic range. Our results demonstrate that MFBBs with different designs can be controlled and combined on a single FCB. Our novel modular approach to operating an automated microfluidic system for parallelized cell culture will enable greater experimental flexibility and facilitate the cooperation of different chips from different labs.

Lactate clearance metrics are not superior to initial lactate in predicting mortality in trauma.

PURPOSE: Despite the availability of different lactate clearance (LC) metrics for clinical use, it remains unknown which metric is superior as a clinical predictor for outcome, particularly in trauma patients. This retrospective study compared four previously described metrics of LC and examined the association between LC and outcome in trauma patients. METHODS: Lactate values of trauma patients admitted to a level I trauma center between 2010 and 2013 were retrieved from patient records. LC was calculated according to Huckabee, Regnier et al., Billeter et al. and Zhang et al. Patients were categorized as isolated traumatic brain injury (TBI), trauma with TBI, and trauma without TBI. The primary study outcome was in-hospital mortality. RESULTS: 367 trauma patients were eligible for LC calculation. Only LC by Zhang et al. [area under the curve (AUC) > 0.622, p < 0.01], and Billeter et al. (AUC > 0.616, p < 0.05) were predictive for mortality in trauma patients with and without TBI. However, both were equally prognostic as the initial lactate value for in-hospital mortality. The prognostic value of initial lactate and lactate clearance for in-hospital mortality were not found to differ between isolated TBI, polytrauma with TBI, and trauma without TBI. CONCLUSIONS: LC metrics based on the methods of Zhang et al. and Billeter et al. predicted mortality in trauma patients, and their prognostic value did not differ between patients with and without TBI. However, initial lactate value was equally prognostic as these LC metrics. Our findings suggest that a single initial lactate measurement may be a more clinically useful tool to predict mortality than the calculation of lactate clearance.

Towards microfluidic sperm refinement: impedance-based analysis and sorting of sperm cells.

The use of high quality semen for artificial insemination in the livestock industry is essential for successful outcome. Insemination using semen with a high number of sperm cells containing morphological defects has a negative impact on fertilization outcome. Therefore, semen with a high number of these abnormal cells is discarded in order to maintain high fertilization potential, resulting in the loss of a large number of morphologically normal sperm cells (up to 70-80% of original sample). A commonly occurring morphological sperm anomaly is the cytoplasmic droplet on the sperm flagella. Currently, no techniques are available to extract morphologically normal sperm cells from rejected samples. Therefore, we aim to develop a microfluidic setup which is able to detect and sort morphologically normal sperm cells label-free and non-invasively. In a proof-of-concept experiment, differential impedance measurements were used to detect the presence of cytoplasmic droplets on sperm flagella, which was quantified by calculating the area under the curve (AUC) of the corresponding impedance peaks. A receiver operating characteristic curve of this electrical analysis method showed the good predictive power of this analysis method (AUC value of 0.85). Furthermore, we developed a label-free cell sorting system using LabVIEW, which is capable of sorting sperm cells based on impedance. In a proof-of-concept experiment, sperm cells and 3 µm beads were sorted label-free and non-invasively using impedance detection and dielectrophoresis sorting. These experiments present our first attempt to perform sperm refinement using microfluidic technology.

The letter digit substitution test: demographic influences and regression-based normative data for school-aged children.

The Letter Digit Substitution Test (LDST) was administered to a sample of N=296 healthy children (aged between 8.03 and 15.87). The aim of the present study was to evaluate the impact of age, gender, and parental educational level on LDST performance and to establish demographically corrected normative data. The results showed that the relationship between age and LDST performance was curvilinear (i.e., improvements in test performance were more pronounced for younger children than for older children) and was moderated by gender (i.e., the gender differences were small at younger ages but increased as a function of age, with girls outperforming boys). Moreover, children who had parents with a higher level of education outperformed their counterparts who had parents with a lower level of education. Regression-based normative LDST data were established, and an automatic scoring program was provided.

Total loss and distribution of nitrogen and phosphorus in the outdoor run of organic laying hens.

1. The objective of this study was to determine the level and variation of the total mass, and load of nitrogen (N) and phosphorus (P) excreted into the outdoor run of organic egg production systems. 2. Three egg production farms with an aviary system and an outdoor run were selected for this study. Four measurements, one per season, were executed on each farm. 3. Mean content of N and P of a manure dropping was 14·0 g N kg⁻¹ and 3·12 g P kg⁻¹, mean mass of a dropping was 6·36 g and mean dry matter content of a dropping was 238 g kg⁻¹. Mean rate of excretion in the outdoor run was 2·99 droppings per hen per h. Mean percentage of hens outside during the time the outdoor run could be accessed was lowest on Farm 1 (1·7%), highest on Farm 2 (16·0%), and intermediate on Farm 3 (7·1%). 4. On all farms an exponential decrease of the number of hens and of the load of N and P with increasing distance from the hen house was found. Load of N exceeded the fertilisation standard (of 170 kg ha⁻¹y⁻¹) in the region at a 0 to 19 m distance from the hen house on Farm 1, 0 to 146 m on Farm 2 and 0 to 52 m on Farm 3. 5. It is concluded that the husbandry system should be redesigned to solve the problem of overloading, unwanted loss of N and P to the environment and loss of N and P from the organic production cycle.

Divergent composition but similar function of soil food webs of individual plants: plant species and community effects.

Soils are extremely rich in biodiversity, and soil organisms play pivotal roles in supporting terrestrial life, but the role that individual plants and plant communities play in influencing the diversity and functioning of soil food webs remains highly debated. Plants, as primary producers and providers of resources to the soil food web, are of vital importance for the composition, structure, and functioning of soil communities. However, whether natural soil food webs that are completely open to immigration and emigration differ underneath individual plants remains unknown. In a biodiversity restoration experiment we first compared the soil nematode communities of 228 individual plants belonging to eight herbaceous species. We included grass, leguminous, and non-leguminous species. Each individual plant grew intermingled with other species, but all plant species had a different nematode community. Moreover, nematode communities were more similar when plant individuals were growing in the same as compared to different plant communities, and these effects were most apparent for the groups of bacterivorous, carnivorous, and omnivorous nematodes. Subsequently, we analyzed the composition, structure, and functioning of the complete soil food webs of 58 individual plants, belonging to two of the plant species, Lotus corniculatus (Fabaceae) and Plantago lanceolata (Plantaginaceae). We isolated and identified more than 150 taxa/groups of soil organisms. The soil community composition and structure of the entire food webs were influenced both by the species identity of the plant individual and the surrounding plant community. Unexpectedly, plant identity had the strongest effects on decomposing soil organisms, widely believed to be generalist feeders. In contrast, quantitative food web modeling showed that the composition of the plant community influenced nitrogen mineralization under individual plants, but that plant species identity did not affect nitrogen or carbon mineralization or food web stability. Hence, the composition and structure of entire soil food webs vary at the scale of individual plants and are strongly influenced by the species identity of the plant. However, the ecosystem functions these food webs provide are determined by the identity of the entire plant community.

Characterization of picosecond pulse nonlinear propagation in chalcogenide As(2)S(3) fiber.

We characterize the nonlinear propagation of picosecond pulses in chalcogenide As(2)S(3) single-mode fiber using a pump-probe technique. The cross-phase modulation (XPM)-induced sideband broadening and stimulated Raman scattering (SRS)-induced sideband amplification are measured in order to map out the Raman gain spectrum of this glass across the C-band. We extract the Raman response function from the Raman gain spectrum and determine the power and polarization dependence of the SRS. In contrast to previous work using As(2)Se(3) fiber, we find that the As(2)S(3) fiber does not suffer from large two-photon absorption (TPA) in the wavelength range of the telecommunications band. We achieved a 20 dB peak Raman gain at a Stokes shift of 350 cm(-1) in a 205 mm length of As(2)S(3) single-mode fiber. The Raman gain coefficient is estimated to be 4.3x10(-12) m/W and the threshold pump peak power is estimated to be 16.2 W for the 205 mm As(2)S(3) fiber. We also demonstrate that we can infer the dispersion of the As(2)S(3) fiber and justify the Raman response function by comparing simulation and experimental results.

A putative mechanism for bog patterning.

The surface of bogs commonly shows various spatial vegetation patterning. Typical are "string patterns" consisting of regular densely vegetated bands oriented perpendicular to the slope. Here, we report on regular "maze patterns" on flat ground, consisting of bands densely vegetated by vascular plants in a more sparsely vegetated matrix of nonvascular plant communities. We present a model reproducing these maze and string patterns, describing how nutrient-limited vascular plants are controlled by, and in turn control, both hydrology and solute transport. We propose that the patterns are self-organized and originate from a nutrient accumulation mechanism. In the model, this is caused by the convective transport of nutrients in the groundwater toward areas with higher vascular plant biomass, driven by differences in transpiration rate. In a numerical bifurcation analysis we show how the maze patterns originate from the spatially homogeneous equilibrium and how this is affected by changes in rainfall, nutrient input, and plant properties. Our results confirm earlier model results, showing that redistribution of a limiting resource may lead to fine-scale facilitative and coarse-scale competitive plant interactions in different ecosystems. Self-organization in ecosystems may be a more general phenomenon than previously thought, which can be mechanistically linked to scale-dependent facilitation and competition.

Contrast in the efficacy of hDAF mouse hearts between ex vivo perfusion and transplantation into primates.

In recent experiments, in which we compared hDAF transgenic rat hearts perfused with 15% human serum in the Langendorff device and hDAF rat hearts transplanted into cynomolgus monkeys, we demonstrated that in the ex vivo heart perfusion model both homozygous and heterozygous hDAF hearts survived longer as nontransgenic controls. Surprisingly, we found that only homozygous hDAF hearts were protected against hyperacute rejection in vivo. The first aim of this study was to determine whether perfusion of mouse hearts with higher human serum concentrations or human blood might explain some of the differences found in survival time of the recently performed experiments with rat heart xenografts. Secondly, we investigated whether the observed differences in survival times of rat xenografts between in vivo and ex vivo transplantation would also hold for mouse hearts transgenic for hDAF. An ex vivo model was used to perfuse hDAF mouse hearts and controls with human serum or blood, and hDAF transgenic hearts and controls were transplanted into cynomolgus monkeys. hDAF transgenic mouse hearts survived significantly longer than their controls when perfused with 15% human serum, but no difference was found when 30% human serum was used, or when these hearts were transplanted into cynomolgus monkeys. However, in both the in vivo and ex vivo models the amount of PMNs adhering to the vascular endothelium was significantly lower in hDAF transgenes as compared with their controls. In conclusion, in the ex vivo situation, the efficacy of hDAF transgenesis in preventing HAR is limited by serum complement concentration.

The ergonomics of flight management systems: fixing holes in the cockpit certification net.

Recent air traffic control regulations mandate the installation of computer-based flight management systems in airliners across Europe. Integrating and certifying add-on cockpit systems is a long and costly process, which in its current form cannot meaningfully address ergonomics aspects. Two levels of problems occur: add-on systems carry many "classic" HCI failures, which could easily be addressed with modified certification requirements. Further, adding new technology changes practice, creates new skill and knowledge demands and produces new forms of error, which are more difficult to assess in advance. However, one innovative certification approach for add-on cockpit systems, based on the use of a representative population of user pilots, was found to be promising. This method minimizes the subjective bias of individual pilots in addition to defining pass/fail criteria in an operational environment.

Skin equivalent: an attractive model to evaluate early melanoma metastasis.

The growth patterns and morphological phenotype of four human melanoma cell lines with different metastatic potentials were investigated in submerged and in air-exposed (skin equivalent) keratinocyte-melanoma cell co-cultures. In contrast to the submerged co-cultures, all four cell lines formed sharply demarcated tumour cell nests within the epidermal compartment of the skin equivalent model, with the morphology highly mimicking the in vivo situation. Differences among the melanoma cell lines tested were observed with respect to the number of clusters formed and the ability to exhibit invasive growth. Only the two metastatic cell lines were able to invade the dermal compartment. Screening of cellular adhesion molecules revealed that the expression patterns in different cell lines were heterogeneous and remained unchanged during the whole culture period, irrespective of whether the melanoma cells were located in the epidermal or dermal compartment. A correlation was found between expression of a lower number of different cellular adhesion molecules and the ability to acquire invasive growth capability. Our results indicate that melanoma cells exhibit a heterogeneous growth behaviour when co-cultured with human keratinocytes, and the air-exposed skin equivalent model was shown to be suitable for studying differences in growth patterns and potential invasive behaviour.

Mycosis fungoides: disease evolution and prognosis of 309 Dutch patients.

OBJECTIVES: To determine the disease course of Dutch patients with mycosis fungoides and to define factors related to disease progression and survival. DESIGN: A multicenter, 13-year, retrospective cohort analysis. SETTING: Eight dermatology departments collaborating in the Dutch Cutaneous Lymphoma Group. PATIENTS: Three hundred nine patients with mycosis fungoides registered between October 1985 and May 1997, including 89 patients with limited patches or plaques (stage Ia), 135 with generalized patches or plaques (stage Ib), 46 with skin tumors (stage Ic), 18 with enlarged but uninvolved lymph nodes (stage II), 18 with lymph node involvement (stage III), and 3 with visceral involvement (stage IV). MAIN OUTCOME MEASURES: Response to initial treatment, sustained complete remission, actuarial disease progression, and overall and disease-specific survival per clinical stage. RESULTS: The median follow-up was 62 months (range, 1-113 months). For the entire group, the actuarial overall and disease-specific survival was 80% and 89% at 5 years, and 57% and 75% at 10 years, respectively. The actuarial 5-year disease-specific survival of patients with stage Ia, Ib, and Ic disease was 100%, 96%, and 80%, respectively, and only 40% for patients with stage III disease. Using multivariate analysis, the presence of extracutaneous disease, the type and extent of skin involvement, the response to initial treatment, and the presence of follicular mucinosis were independently associated with higher disease progression and mortality rates. The calculated risks of disease progression at 5 and 10 years gradually increased from 4% to 10% for those with stage Ia disease, from 21% to 39% for those with stage Ib disease, and from 32% to 60% for those with stage Ic disease; for those with stage III disease, the risk remained at 70% at 5 and 10 years. The overall risk of disease progression at 5 and 10 years was 24% and 38%, respectively, for the total study group. CONCLUSION: At least within the first 10 years after diagnosis, disease progression and mycosis fungoides-related mortality occur in only a subset of patients generally presenting with advanced disease.

Modeling daily gas exchange of a Douglas-fir forest: comparison of three stomatal conductance models with and without a soil water stress function.

Modeling stomatal conductance is a key element in predicting tree growth and water use at the stand scale. We compared three commonly used models of stomatal conductance, the Jarvis-Loustau, Ball-Berry and Leuning models, for their suitability for incorporating soil water stress into their formulation, and for their performance in modeling forest ecosystem fluxes. We optimized the parameters of each of the three models with sap flow and soil water content data. The optimized Ball-Berry model showed clear relationships with air temperature and soil water content, whereas the optimized Leuning and Jarvis-Loustau models only showed a relationship with soil water content. We conclude that use of relative humidity instead of vapor pressure deficit, as in the Ball-Berry model, is not suitable for modeling daily gas exchange in Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) in the Speulderbos forest near the village of Garderen, The Netherlands. Based on the calculated responses to soil water content, we linked a model of forest growth, FORGRO, with a model of soil water, SWIF, to obtain a forest water-balance model that satisfactorily simulated carbon and water (transpiration) fluxes and soil water contents in the Douglas-fir forest for 1995.