Assessing the inter-annual variability of separation distances around odour sources to protect the residents from odour annoyance.

In recent years, there has been a growing concern about potential impacts on public health and wellbeing due to exposure to environmental odour. Separation distances between odour-emitting sources and residential areas can be calculated using dispersion models, as a means of protecting the neighbourhood from odour annoyance. This study investigates the suitability of using one single year of meteorological input data to calculate reliable direction-dependent separation distances. Accordingly, we assessed and quantified the inter-annual variability of separation distances at two sites with different meteorological conditions, one in Brazil and the other in Austria. A 5-year dataset of hourly meteorological observations was used for each site. Two odour impact criteria set in current regulations were selected to explore their effect on the separation distances. The coefficient of variation was used as a statistical measure to characterise the amount of annual variation. Overall, for all scenarios, the separation distances had a low degree of inter-annual variability (mean coefficient of variation values from 8% to 21%). Reasonable agreements from year to year were therefore observed at the two sites under investigation, showing that one year of meteorological data is a good compromise to achieve reliable accuracy. This finding can provide a more cost-effective solution to calculate separation distances in the vicinity of odour sources.

Are the simplified methods to estimate Ki in 18F-FDG PET studies feasible in clinical routine? Comparison between three simplified methods.

BACKGROUND: The aim of this study was to verify the feasibility, in clinical practice, of three simplified methods (Hunter, Sadato and another one proposed by our group) to calculate Ki and MRglu of 18F-FDG, comparing the results with those derived by the linear regression (LR) method (considered the golden standard), and also with SUV. METHODS: Forty-five patients (32males, mean age 69±9years) with non-small-cell-lung cancer prospectively enrolled, underwent dynamic 18F-FDG PET-CT over the thorax. Ki was estimated as follows: from a static acquisition and performing one venous blood sampling using the Hunter method; multiplying the SUV for the average plasma clearance rate (kP(T)) and for the initial distribution volume (V0bw) without performing any blood sampling using the Sadato method; multiplying the SUV for a factor F (which encompasses the mean value of haematocrit and plasma volume, both according to patient's sex) without performing any blood sample using ours method. Wilcoxon signed rank and coefficient of determination (R2) were used for statistical analysis. RESULTS: No significant difference was observed between the Ki and MRglu estimated by all three simplified methods and the Ki and MRglu estimated by LR. The highest P values and the lower values of mean differences were observed with our method compared with LR: Ki=0.0392±0.0178 min-1 vs. Ki=0.0392±0.0202 min-1 (P=0.897, MD=0.0001 min-1), respectively; MRglu= 4.47±2.23 ml/min/100g vs. MRglu= 4.43±2.38 ml/min/100g (P=0.839, MD= -0.0373 mL/min/100g), respectively. The highest correlation was observed between the Ki estimated by both Hunter and our methods and the Ki estimated by LR: R2=0.87, R2=0.86, respectively. A good correlation (R2=0.83) was observed between SUV and Ki estimated by LR. CONCLUSIONS: These three simplified methods represent a valid alternative to the more invasive and complex full kinetic analysis. Their "pros" are: the non-invasiveness, the feasibility, the good correlation with the golden standard; their "cons" is that full kinetic analysis provides highest accuracy in Ki determination. Therefore, in clinical oncology routine, the nuclear physicians can choose among different simplified methods especially for monitoring the response to treatment, for tumour grading, and for prognostic stratification, letting the full kinetic analysis to specific centre/studies.

Toward a rational design of surface textures promoting endothelialization.

The safe integration of cardiovascular devices requires the sustainable coverage of their luminal surface by endothelial cells (ECs). The engineering of active surface textures has the potential to coordinate cellular adhesion and migration under the action of hemodynamic forces. We define a paradigm to rationally design textures maximizing EC activities as a function of the applied stresses. This is based on harnessing the adhesions established by ECs through fine-tuning of the vertical extend of the underlying surface nanotopography.

Investigation of rainwater contamination sources in the southern part of Brazil.

Rainwater quality is influenced by air pollutants and can affect sensitive ecosystems. This study was conducted to identify the sources of rainwater contamination in a receptor investigated in the southern part of Brazil. A total of 22 rainwater samples were collected at Florianópolis, Brazil. The sampling station is influenced by continental emissions (soil resuspension, traffic emissions and combustion) and marine aerosols. Over the sampling period, the average pH and electrical conductivity (EC) of the precipitation was found to be 4.97 +/- 0.41 and 14.68 microS cm(-1) +/- 13.47, respectively. In addition topH and EC, ions and trace metals in the collected rainwater were quantified. The results were investigated by a combination of techniques including principal component analysis (PCA), a back trajectory model and other statistical and graphical interpretation methodologies. A PCA showed that Cl(-), Na+, Mg2+ and part of the K+ and SO4(2-) content were mainly contributed by marine aerosols, whereas the contribution from continental sources (combustion, traffic emissions and other urban activities) was dominant in the content of NO3(-) and part of the SO4(2-) and Mn content. Soil resuspension was responsible for the concentrations of most of the trace metals (apart from Mn) and Ca2+ in the rainwater. An inverse correlation among the elemental concentrations, amount of rainfall and wind speed was observed. The northern transport pathway was identified as being associated with high concentrations of NO3(-) and slightly decreased pH values. However, the low standard deviation observed for the pH values during the sampling campaign also showed a small variation in the data, suggesting that the acidity is most probably being constantly sourced from a natural origin, such as organic acids.

A review of plastic debris in the South American Atlantic Ocean coast - Distribution, characteristics, policies and legal aspects.

The presence of plastics in the oceans has already become a pervasive phenomenon. Marine pollution by plastics surpasses the status of an emerging threat to become a well-established environmental problem, boosting research on this topic. However, despite many studies on the main seas and oceans, it is necessary to compile information on the South American Atlantic Ocean Coast to identify the lack of research and expand knowledge on marine plastic pollution in this region. Accordingly, this paper conducted an in-depth review of monitoring methods, sampling, and identification of macroplastics and microplastics (MPs) in water, sediments, and biota, including information on legal requirements from different countries as well as non-governmental initiatives. Brazil was the country with the highest number of published papers, followed by Argentina. MPs accounted for 75 % of the papers selected, with blue microfibers being the most common morphology, whereas PE and PP were the most abundant polymers. Also, a lack of standardization in the methodologies used was identified; however, the sites with the highest concentrations of MPs were the Bahía Blanca Estuary (Argentina), Guanabara Bay (Brazil), and Todos os Santos Bay (Brazil), regardless of the method applied. Regarding legislation, Uruguay and Argentina have the most advanced policies in the region against marine plastic pollution due to their emphasis on the life cycle and the national ban on certain single-use plastics. Therefore, considering its content, this expert review can be useful to assist researchers dealing with plastic pollution along the South American Atlantic Ocean Coast.

Pseudidotheaarmata sp. n., a new isopod of the genus Pseudidothea (Crustacea, Malacostraca, Isopoda) from the Atlantic sector of the Southern Ocean.

BACKGROUND: In the framework of the British Antarctic Survey (BAS) Expedion JR 15005 SO-AntEco, held in February-March 2016, the South Orkney Islands seafloor was sampled in order to investigate the distribution and composition of benthic communities around the area. NEW INFORMATION: A new species of the genus Pseudidothea Ohlin, 1901 is described from the Burdwood Bank area (South Orkney Islands). It has been collected during the SO-AntEco JR15005 RRS James Clark Ross expedition under the lead of the British Antarctic Survey (BAS). The new species, Pseudidotheaarmata sp. n., is very similar to P.scutata (Stephensen, 1947); however, it is characterised by peculiar supra-ocular spines and a different tubercular pattern. The study of the species of the Pseudidothea helps to better understand the diversity of the Pseudidotheidae in the Southern Ocean.

Proposed search for a fourth neutrino with a PBq antineutrino source.

Several observed anomalies in neutrino oscillation data can be explained by a hypothetical fourth neutrino separated from the three standard neutrinos by a squared mass difference of a few eV(2). We show that this hypothesis can be tested with a PBq (ten kilocurie scale) (144)Ce or (106)Ru antineutrino beta source deployed at the center of a large low background liquid scintillator detector. In particular, the compact size of such a source could yield an energy-dependent oscillating pattern in event spatial distribution that would unambiguously determine neutrino mass differences and mixing angles.

Abundance data of benthic peracarid crustaceans from the South Atlantic and Southern Ocean.

Peracarid data were collected in the Southern Ocean and South Atlantic Ocean. Sampling was performed during nine different expeditions on board of RRS James Clark Ross and RV Polarstern, using epibenthic sledges (EBS) at depth ranging between 160-6348 m at 109 locations. The correlation between environmental variables and peracarid abundance was investigated. Abundance data comprise a total of 128570 peracarids (52366 were amphipods, 28516 were cumaceans, 36142 isopods, 5676 mysidaceans and 5870 were tanaidaceans). The presented data are useful to investigate the composition and abundance patterns of peracarid orders at a wide depth range and spatial scale in the Southern Ocean. They can also be reused to compare their abundance with that of other taxa in broader ecological surveys.

An analysis of error propagation in AERMOD lateral dispersion using Round Hill II and Uttenweiller experiments in reduced averaging times.

Dispersion modelling was proved by researchers that most part of the models, including the regulatory models recommended by the Environmental Protection Agency of the United States (AERMOD and CALPUFF), do not have the ability to predict under complex situations. This article presents a novel evaluation of the propagation of errors in lateral dispersion coefficient of AERMOD with emphasis on estimate of average times under 10 min. The sources of uncertainty evaluated were parameterizations of lateral dispersion ([Formula: see text]), standard deviation of lateral wind speed ([Formula: see text]) and processing of obstacle effect. The model's performance was tested in two field tracer experiments: Round Hill II and Uttenweiller. The results show that error propagation from the estimate of [Formula: see text] directly affects the determination of [Formula: see text], especially in Round Hill II experiment conditions. After average times are reduced, errors arise in the parameterization of [Formula: see text], even after observation assimilations of [Formula: see text], exposing errors on Lagrangian Time Scale parameterization. The assessment of the model in the presence of obstacles shows that the implementation of a plume rise model enhancement algorithm can improve the performance of the AERMOD model. However, these improvements are small when the obstacles have a complex geometry, such as Uttenweiller.

A review of odour impact criteria in selected countries around the world.

Exposure to environmental odour can result in annoyance, health effects and depreciation of property values. Therefore, many jurisdictions classify odour as an atmospheric pollutant and regulate emissions and/or impacts from odour generating activities at a national, state or municipal level. In this work, a critical review of odour regulations in selected jurisdictions of 28 countries is presented. Individual approaches were identified as: comparing ambient air odour concentration and individual chemicals statistics against impact criteria (maximum impact standard); using fixed and variable separation distances (separation distance standard); maximum emission rate for mixtures of odorants and individual chemical species (maximum emission standard); number of complaints received or annoyance level determined via community surveys (maximum annoyance standard); and requiring use of best available technologies (BAT) to minimize odour emissions (technology standard). The comparison of model-predicted odour concentration statistics against odour impact criteria (OIC) is identified as one of the most common tools used by regulators to evaluate the risk of odour impacts in planning stage assessments and is also used to inform assessment of odour impacts of existing facilities. Special emphasis is given to summarizing OIC (concentration percentile and threshold) and the manner in which they are applied. The way short term odour peak to model time-step mean (peak-to-mean) effects is also captured. Furthermore, the fundamentals of odorant properties, dimensions of nuisance odour, odour sampling and analysis methods and dispersion modelling guidance are provided. Common elements of mature and effective odour regulation frameworks are identified and an integrated multi-tool strategy is recommended.

Photocatalytic oxidation of H2S in the gas phase over TiO2-coated glass fiber filter.

To promote the photocatalytic oxidation (PCO) of hydrogen sulfide (H2S) in the gas phase, TiO2-coated glass fiber filters were packed in an annular photoreactor. Glass fibers coated with TiO2 thin films were characterized structurally and morphologically by field emission gun scanning electron microscopy (FEG-SEM), scanning electron microscopy (SEM) coupled with energy-dispersive X-ray spectroscopy (EDS) and X-ray diffractometry (XRD). Flow rate and H2S inlet concentration were evaluated to determine the performance of the reactor. Removal efficiencies up to 99% were achieved for flow rate of 25 L h(-1) (residence time of 121 s) and H2S inlet concentration from 12 to 14 ppmv. The long-term experiment presented H2S removal of 89% for 16 h. After 28 h of continuous use, H2S degradation was observed at 64%, which suggests that the photocatalyst was losing activity due to deactivation. Moreover, the kinetics of the PCO of H2S according to the Langmuir-Hinshelwood (L-H) approach along with the mass balance of a plug-flow reactor was modeled. The reaction constant (k) was calculated at approximately 10.5 µmol m(-3) s(-1) and the adsorption constant (K) of approximately 5263 m(-3) mol with linearity (R2) of 0.98.

Potential Usefulness of 99mTc-DMSA for Radio-Guided Surgery in Pediatric Renal Dysplasia.

We report a case of an ectopic/hypoplastic kidney removed by radio-guided surgery. A 7-year-old girl, with a history of vaginal drainage of urine, underwent renal scintigraphy with Tc-DMSA. SPECT/CT revealed a focal uptake in the pelvis, corresponding to hypoplastic kidney as confirmed by MRI. Based on SPECT/CT findings, the patient underwent laparoscopic surgery, using Tc-DMSA scan to help the surgeon to detect the small ectopic kidney. Intraoperatory histological report confirmed the renal origin of the specimen.

Diagnostic Performance of (18)F-Fluorodeoxyglucose in 162 Small Pulmonary Nodules Incidentally Detected in Subjects Without a History of Malignancy.

BACKGROUND: Solitary pulmonary nodule (SPN) still represents a diagnostic challenge. The aim of our study was to evaluate the diagnostic performance of (18)F-fluorodeoxyglucose positron emission tomography-computed tomography in one of the largest samples of small SPNs, incidentally detected in subjects without a history of malignancy (nonscreening population) and undetermined at computed tomography. METHODS: One-hundred and sixty-two small (>0.8 to 1.5 cm) and, for comparison, 206 large nodules (>1.5 to 3 cm) were retrospectively evaluated. Diagnostic performance of (18)F-fluorodeoxyglucose visual analysis, receiver-operating characteristic (ROC) analysis for maximum standardized uptake value (SUVmax), and Bayesian analysis were assessed using histology or radiological follow-up as a golden standard. RESULTS: In 162 small nodules, (18)F-fluorodeoxyglucose visual and ROC analyses (SUVmax = 1.3) provided 72.6% and 77.4% sensitivity and 88.0% and 82.0% specificity, respectively. The prevalence of malignancy was 38%; Bayesian analysis provided 78.8% positive and 16.0% negative posttest probabilities of malignancy. In 206 large nodules (18)F-fluorodeoxyglucose visual and ROC analyses (SUVmax = 1.9) provided 89.5% and 85.1% sensitivity and 70.8% and 79.2% specificity, respectively. The prevalence of malignancy was 65%; Bayesian analysis provided 85.0% positive and 21.6% negative posttest probabilities of malignancy. In both groups, malignant nodules had a significant higher SUVmax (p < 0.0001) than benign nodules. Only in the small group, malignant nodules were significantly larger (p = 0.0054) than benign ones. CONCLUSIONS: (18)F-fluorodeoxyglucose can be clinically relevant to rule in and rule out malignancy in undetermined small SPNs, incidentally detected in nonscreening population with intermediate pretest probability of malignancy, as well as in larger ones. Visual analysis can be considered an optimal diagnostic criterion, adequately detecting a wide range of malignant nodules with different metabolic activity.

Influência de diferentes tipos de nanopartículas de prata na biodegradação da fração orgânica de resíduos sólidos urbanos / Influence of different types of silver nanoparticles on the biodegradation of the organic fraction of municipal solid waste

RESUMO Os resíduos sólidos urbanos (RSU) contêm aproximadamente 50% de fração orgânica (FO), que, por meio de processos de biodegradação em aterros sanitários, resultam em lixiviados e biogás. A degradação anaeróbia ocorre impulsionada por processos microbianos realizados por um consórcio complexo de micro-organismos, responsáveis pelas diferentes fases. Por outro lado, observa-se que as nanopartículas (NP), especialmente as de prata (NPsAg), vêm sendo cada vez mais utilizadas em novos produtos. Assim, os RSU podem apresentar materiais nanoparticulados, que, depositados em aterros sanitários, se comportam de maneira ainda pouco conhecida, podendo influenciar os processos de biodegradação. Os objetivos do presente trabalho são analisar e verificar o potencial metanogênico de resíduos sólidos orgânicos com e sem a interferência de diferentes concentrações e tipos de NPsAg. Para isso, foram realizados ensaios do potencial biometanogênico com as concentrações de 5, 15, 45, 50, 150 e 450 mg NPsAg.kg−1 RSU e comparados com a amostra padrão (branco). Diferentes tipos de nanopartículas foram utilizados: sólido (em pó), solução em base aquosa e em base alcoólica com álcool polivinílico (PVA). Os resultados mostram que, em concentrações altas (150 e 450 mg NPsAg.kg−1), as NPsAg tendem a diminuir a atividade dos micro-organismos metanogênicos. Finalmente, a modelagem dos resultados mostrou que o modelo logístico se ajusta bem aos dados experimentais. A regressão linear entre os parâmetros desse modelo e a concentração de NPsAg demonstram o decréscimo dos parâmetros com o aumento na concentração desses compostos.

ABSTRACT Municipal solid waste (MSW) contains approximately 50% of organic fraction (OF), which through landfill biodegradation processes results in leachate and biogas. Anaerobic degradation occurs driven by microbial processes, with a complex consortium of microorganisms, responsible for different phases. On the other hand, it is observed that nanoparticles (NPs), specially silver nanoparticles (AgNPs), have been appearing in a large range of new products. The MSW, for example, can present nanoparticulate materials, which, deposited in landfills, behave in a way still little known in science, and that can influence the biodegradation processes. The objective of this work is to analyze and verify the biochemical methanogenic potential of organic solid waste with and without the interference of different concentrations and types of AgNPs. For that purpose, tests of biochemical methane potential (BMP) with the concentrations of 5, 15 and 45, 50, 150 and 450 AgNPs mg.kg−1 were carried out and verified through a standard sample (white). Different types of nanoparticles were used: solid (powder), and two solutions, aqueous and alcoholic (polyvinyl alcohol — PVA). The results showed the interference of AgNPs in the highest concentrations used, considerably decreasing the biological process measured through the activity of microorganisms in the production of methane gas, which occurred in the range between 150 and 450 mg.kg−1 of AgNPs. Finally, the statistical studies and the mathematical modeling of the results showed that the logistic model adjusted well to the experimental data. The linear regression of the model parameters with the concentration of AgNPs demonstrates the decrease in the values of the parameters with the increase in the concentration of these compounds.

A thyroid incidentaloma detected by 18F-choline PET/CT.

We report a case of thyroid incidentaloma detected by 18F-choline PET/CT. A 66-year-old male patient with a history of prostate cancer underwent a 18F-choline PET/CT for restaging. PET/CT revealed a focal area of increased 18F-choline uptake corresponding to a hypodense nodule in the right lobe of the thyroid. Based on PET/CT findings, the patient underwent a ultrasonography guided fine-needle aspiration biopsy which demonstrated the presence of a benign thyroid nodule.

The influence of surface micro-structure on endothelialization under supraphysiological wall shear stress.

Interaction between platelets and artificial materials within cardiovascular devices triggers blood coagulation and represents a frequent adverse response to implant deployment. Avoidance of this interaction is obtained through the generation and sustenance under flow of a confluent and stable endothelial monolayer covering the luminal device surface, altogether defined as the process of endothelialization. Supraphysiological wall shear stress (WSS) levels generated within vascular assist devices (VADs) constitute a major challenge toward endothelialization. Here we report the experimental demonstration that stable endothelialization can be achieved at supraphysiological WSS levels by pure means of appropriate surface micro-structuring. Using a custom-designed flow bioreactor we exposed endothelial monolayers to physiological and supraphysiological WSS levels and investigated the resulting integrity of cell-to-cell junctions, the cell density and the cell polarization. At physiological WSS levels, optimal endothelialization was obtained independently from surface topography. However, at higher WSS levels, only monolayers grown on appropriately micro-structured surfaces preserved optimal integrity. Under these flow conditions, endothelial cells polarized by the contact with the micro-structure and, interestingly, oriented themselves in the direction perpendicular to flow. Such endothelial layers withstood WSS levels exceeding of 100% or more the thresholds detected on flat substrates.

[Future perspectives for diagnostic imaging in urology: from anatomic and functional to molecular imaging]. / Scenari futuri della diagnostica per immagini in urologia: passaggio dall'imaging anatomico e funzionale all'imaging molecolare.

The future approach of diagnostic imaging in urology follows the technological progress, which made the visualization of in vivo molecular processes possible. From anatomo-morphological diagnostic imaging and through functional imaging molecular radiology is reached. Based on molecular probes, imaging is aimed at assessing the in vivo molecular processes, their physiology and function at cellular level. The future imaging will investigate the complex tumor functioning as metabolism, aerobic glycolysis in particular, angiogenesis, cell proliferation, metastatic potential, hypoxia, apoptosis and receptors expressed by neoplastic cells. Methods for performing molecular radiology are CT, MRI, PET-CT, PET-MRI, SPECT and optical imaging. Molecular ultrasound combines technological advancement with targeted contrast media based on microbubbles, this allowing the selective registration of microbubble signal while that of stationary tissues is suppressed. An experimental study was carried out where the ultrasound molecular probe BR55 strictly bound to prostate tumor results in strong enhancement in the early phase after contrast, this contrast being maintained in the late phase. This late enhancement is markedly significant for the detection of prostatic cancer foci and to guide the biopsy sampling. The 124I-cG250 molecular antibody which is strictly linked to cellular carbonic anhydrase IX of clear cell renal carcinoma, allows the acquisition of diagnostic PET images of clear cell renal carcinoma without biopsy. This WG-250 (RENCAREX) antibody was used as a therapy in metastatic clear cell renal carcinoma. Future advancements and applications will result in early cancer diagnosis, personalized therapy that will be specific according to the molecular features of cancer and leading to the development of catheter-based multichannel molecular imaging devices for cystoscopy-based molecular imaging diagnosis and intervention.

On cell separation with topographically engineered surfaces.

BACKGROUND: Topographical modifications of the surface influence several cell functions and can be exploited to modulate cellular activities such as adhesion, migration and proliferation. These complex interactions are cell-type specific, therefore engineered substrates featuring patterns of two or more different topographies may be used to obtain the selective separation of different cell lineages. This process has the potential to enhance the performance of biomedical devices promoting, for example, the local coverage with functional tissues while demoting the onset of inflammatory reactions. FINDINGS & CONCLUSIONS: Here we present a computational tool, based on Monte Carlo simulation, which decouples the contribution of cell proliferation and migration and predicts the cell-separation performance of topographically engineered substrates. Additionally, we propose an optimization procedure to shape the topographically engineered areas of a substrate and obtain maximal cell separation.

A novel 3D integrated platform for the high-resolution study of cell migration plasticity.

Understanding the mechanisms of interstitial cancer migration is of great scientific and medical interest. Creating 3D platforms, conducive to optical microscopy and mimicking the physical parameters (in plane and out of plane) involved in interstitial migration, is a major step forward in this direction. Here, a novel approach is used to directly print free-form, 3D micropores on basal scaffolds containing microgratings optimized for contact guidance. The platforms so formed are validated by monitoring cancer cell migration and micropore penetration with high-resolution optical microscopy. The shapes, sizes and deformability of the micropores are controllable, paving the way to decipher their role in interstitial migration.

Accelerated endothelial wound healing on microstructured substrates under flow.

Understanding and accelerating the mechanisms of endothelial wound healing is of fundamental interest for biotechnology and of significant medical utility in repairing pathologic changes to the vasculature induced by invasive medical interventions. We report the fundamental mechanisms that determine the influence of substrate topography and flow on the efficiency of endothelial regeneration. We exposed endothelial monolayers, grown on topographically engineered substrates (gratings), to controlled levels of flow-induced shear stress. The wound healing dynamics were recorded and analyzed in various configurations, defined by the relative orientation of an inflicted wound, the topography and the flow direction. Under flow perpendicular to the wound, the speed of endothelial regeneration was significantly increased on substrates with gratings oriented in the direction of the flow when compared to flat substrates. This behavior is linked to the dynamic state of cell-to-cell adhesions in the monolayer. In particular, interactions with the substrate topography counteract Vascular Endothelial Cadherin phosphorylation induced by the flow and the wounding. This effect contributes to modulating the mechanical connection between migrating cells to an optimal level, increasing their coordination and resulting in coherent cell motility and preservation of the monolayer integrity, thus accelerating wound healing. We further demonstrate that the reduction of vascular endothelial cadherin phosphorylation, through specific inhibition of Src activity, enhances endothelial wound healing in flows over flat substrates.