Comparative Study of Chemoembolization, Loadable Beads: In vitro Drug Release and Physical Properties of DC Bead and Hepasphere, Loaded with Doxorubicin and Irinotecan

Purpose To characterize in vitro the loadability, physical properties, and release of irinotecan and doxorubicin from two commercially available embolization microspheres. Materials and Methods DC Bead (500-700 μm) and Hepasphere (400-600 μm) microspheres were loaded with either doxorubicin or irinotecan solutions. Drug amount was quantified with spectrophotometry, bead elasticity was measured under compression, and bead size and loading homogeneity were assessed with microscopy image analysis. Drug release was measured over 1-week periods in saline by using a pharmacopeia flow-through method. Results Almost complete drug loading was obtained for both microsphere types and drugs. Doxorubicin-loaded DC Beads maintained their spherical shape throughout the release. In contrast, Hepaspheres showed less homogeneous doxorubicin loading and, after release, some fractured microspheres. Incomplete doxorubicin release was observed in saline over 1 week (27% ± 2 for DC beads and 18% ± 7 for Hepaspheres; P = .013). About 75% of this amount was released within 2.2 hours for both beads. For irinotecan, complete release was obtained for both types of beads, in a sustained manner over 2-3 hours for DC Beads, and in a significantly faster manner as a 7-minute burst for Hepaspheres. Conclusions The two drug-eluting microspheres could be efficiently loaded with both drugs. Incomplete doxorubicin release was attributed to strong drug-bead ionic interactions. Weaker interactions were observed with irinotecan, which led to faster drug release.

Automatic classification of MR scans in Alzheimer's disease.

To be diagnostically useful, structural MRI must reliably distinguish Alzheimer's disease (AD) from normal aging in individual scans. Recent advances in statistical learning theory have led to the application of support vector machines to MRI for detection of a variety of disease states. The aims of this study were to assess how successfully support vector machines assigned individual diagnoses and to determine whether data-sets combined from multiple scanners and different centres could be used to obtain effective classification of scans. We used linear support vector machines to classify the grey matter segment of T1-weighted MR scans from pathologically proven AD patients and cognitively normal elderly individuals obtained from two centres with different scanning equipment. Because the clinical diagnosis of mild AD is difficult we also tested the ability of support vector machines to differentiate control scans from patients without post-mortem confirmation. Finally we sought to use these methods to differentiate scans between patients suffering from AD from those with frontotemporal lobar degeneration. Up to 96% of pathologically verified AD patients were correctly classified using whole brain images. Data from different centres were successfully combined achieving comparable results from the separate analyses. Importantly, data from one centre could be used to train a support vector machine to accurately differentiate AD and normal ageing scans obtained from another centre with different subjects and different scanner equipment. Patients with mild, clinically probable AD and age/sex matched controls were correctly separated in 89% of cases which is compatible with published diagnosis rates in the best clinical centres. This method correctly assigned 89% of patients with post-mortem confirmed diagnosis of either AD or frontotemporal lobar degeneration to their respective group. Our study leads to three conclusions: Firstly, support vector machines successfully separate patients with AD from healthy aging subjects. Secondly, they perform well in the differential diagnosis of two different forms of dementia. Thirdly, the method is robust and can be generalized across different centres. This suggests an important role for computer based diagnostic image analysis for clinical practice.

Caracterização Geológica e Geotécnica das Unidades Litológicas da Cidade da Praia (Santiago - Cabo Verde)

Este trabalho produz uma avaliação das características geológicas e geotécnicas de um território da ilha de Santiago, Cabo Verde, incidindo sobre a cidade da Praia e a área envolvente. Os objectivos principais do trabalho são a representação cartográfica das unidades litológicas, com uma proposta de coluna litológica sintética, a correlação com as unidades geológicas definidas por outros autores, bem como a realização de um conjunto de ensaios in situ e análises laboratoriais visando a caracterização dos parâmetros físicos e mecânicos das diferentes unidades litológicas definidas para a área em estudo. É efectuada uma compilação sobre as principais investigações geológicas realizadas em Cabo Verde, desde o séc. XIX até a actualidade, assim como a descrição das características geológicas da ilha de Santiago e dos principais processos de formação e evolução das formas de relevo e dos recursos naturais (hídricos e solos). A pesquisa incluiu uma revisão dos conceitos descritivos de caracterização dos vulcanitos e vulcanoclastitos presentes e compreendeu a redefinição das unidades litológicas da região, a respectiva representação cartográfica na escala 1: 10.000 (com maior detalhe do que a informação pré-existente) e o levantamento de campo dos principais alinhamentos estruturais. São igualmente apresentados os resultados do tratamento das bandas espectrais de imagens de satélite que suportam a definição das unidades litológicas, bem como dos principais alinhamentos tectónicos que afectam as unidades da área de estudo. Para a caracterização das diferentes unidades litológicas com base nas propriedades in situ, recorreu-se a perfis-tipo, estabelecendo uma descrição das características mineralógicas, petrográficas, texturais e estruturais das materiais presentes. Os materiais foram igualmente ensaiados in situ e amostrados para uma caracterização laboratorial. Os resultados dos ensaios geotécnicos, geomecânicos, mineralógicos e de radioactividade natural permitiram refinar a cartografia proposta e estabelecer parâmetros de comportamento para as 30 unidades litológicas consideradas, a partir de intervalos de valores não homogéneos, contudo representativos da variabilidade observada. Com base nestes resultados são apresentados valores indicativos de aptidão dos materiais, nomeadamente para fundações de edifícios, aterros, construção e potencial hidrogeológico.

Profiling of counterfeit medicines by vibrational spectroscopy

Counterfeit pharmaceutical products have become a widespread problem in the last decade. Various analytical techniques have been applied to discriminate between genuine and counterfeit products. Among these, Near-infrared (NIR) and Raman spectroscopy provided promising results.The present study offers a methodology allowing to provide more valuable information fororganisations engaged in the fight against counterfeiting of medicines.A database was established by analyzing counterfeits of a particular pharmaceutical product using Near-infrared (NIR) and Raman spectroscopy. Unsupervised chemometric techniques (i.e. principal component analysis - PCA and hierarchical cluster analysis - HCA) were implemented to identify the classes within the datasets. Gas Chromatography coupled to Mass Spectrometry (GC-MS) and Fourier Transform Infrared Spectroscopy (FT-IR) were used to determine the number of different chemical profiles within the counterfeits. A comparison with the classes established by NIR and Raman spectroscopy allowed to evaluate the discriminating power provided by these techniques. Supervised classifiers (i.e. k-Nearest Neighbors, Partial Least Squares Discriminant Analysis, Probabilistic Neural Networks and Counterpropagation Artificial Neural Networks) were applied on the acquired NIR and Raman spectra and the results were compared to the ones provided by the unsupervised classifiers.The retained strategy for routine applications, founded on the classes identified by NIR and Raman spectroscopy, uses a classification algorithm based on distance measures and Receiver Operating Characteristics (ROC) curves. The model is able to compare the spectrum of a new counterfeit with that of previously analyzed products and to determine if a new specimen belongs to one of the existing classes, consequently allowing to establish a link with other counterfeits of the database.

Programa d'anàlisi d'imatges de provetes metal·logràfiques

L’objectiu del present TFM és explorar les possibilitats del programa matemàtic MATLAB i la seva eina Entorn de Disseny d’Interfícies Gràfiques d’Usuari (GUIDE), desenvolupant un programa d’anàlisi d’imatges de provetes metal·logràfiques que es pugui utilitzar per a realitzar pràctiques de laboratori de l’assignatura Tecnologia de Materials de la titulació de Grau en Enginyeria Mecatrònica que s’imparteix a la Universitat de Vic. Les àrees d’interès del treball són la Instrumentació Virtual, la programació MATLAB i les tècniques d’anàlisi d’imatges metal·logràfiques. En la memòria es posa un èmfasi especial en el disseny de la interfície i dels procediments per a efectuar les mesures. El resultat final és un programa que satisfà tots els requeriments que s’havien imposat en la proposta inicial. La interfície del programa és clara i neta, destinant molt espai a la imatge que s’analitza. L’estructura i disposició dels menús i dels comandaments ajuda a que la utilització del programa sigui fàcil i intuïtiva. El programa s’ha estructurat de manera que sigui fàcilment ampliable amb altres rutines de mesura, o amb l’automatització de les rutines existents. Al tractar-se d’un programa que funciona com un instrument de mesura, es dedica un capítol sencer de la memòria a mostrar el procediment de càlcul dels errors que s’ocasionen durant la seva utilització, amb la finalitat de conèixer el seu ordre de magnitud, i de saber-los calcular de nou en cas que variïn les condicions d’utilització. Pel que fa referència a la programació, malgrat que MATLAB no sigui un entorn de programació clàssic, sí que incorpora eines que permeten fer aplicacions no massa complexes, i orientades bàsicament a gràfics o a imatges. L’eina GUIDE simplifica la realització de la interfície d’usuari, malgrat que presenta problemes per tractar dissenys una mica complexos. Per altra banda, el codi generat per GUIDE no és accessible, cosa que no permet modificar manualment la interfície en aquells casos en els que GUIDE té problemes. Malgrat aquests petits problemes, la potència de càlcul de MATLAB compensa sobradament aquestes deficiències.

A rapid and effective vignetting correction for quantitative microscopy

Images acquired using optical microscopes are inherently subject to vignetting effects due to imperfect illumination and image acquisition. However, such vignetting effects hamper accurate extraction of quantitative information from biological images, leading to less effective image segmentation and increased noise in the measurements. Here, we describe a rapid and effective method for vignetting correction, which generates an estimate for a correction function from the background fluorescence without the need to acquire additional calibration images. We validate the usefulness of this algorithm using artificially distorted images as a gold standard for assessing the accuracy of the applied correction and then demonstrate that this correction method enables the reliable detection of biologically relevant variation in cell populations. A simple user interface called FlattifY was developed and integrated into the image analysis platform YeastQuant to facilitate easy application of vignetting correction to a wide range of images.

Soil structure transformations from ferralic to nitic horizons on a toposequence in southeastern Brazil

The soil structure transformation from ferralic to nitic horizons was studied in a toposequence on quaternary red clayey sediments and diabase in Piracicaba (SP), Brazil. Morphological and micromorphological studies, image analysis, soil water characteristic curves and monitoring of (total) soil water potential head were used. The presence of polyconcave vughs, clayskins and planar voids shows that the vertical and lateral transition and structural transformation from ferralic to nitic horizons is given by the coalescence of the microaggregates, probably due to tensions created in a drier period in the past. Changes to a more humid climate with a defined dry season and alternate drying and wetting cycles resulted in the fissuration of the previously coalesced material, forming polyhedral aggregates and microaggregates. Simultaneously, clay illuviation filled the voids and together with the compacting action of the biological activity of these soils contributed to the coalescence of microaggregates.

Quantification of the local heartwall motion by magnetic resonance myocardial tagging.

Sophisticated magnetic resonance tagging techniques provide powerful tools for the non-invasive assessment of the local heartwall motion towards a deeper fundamental understanding of local heart function. For the extraction of motion data from the time series of magnetic resonance tagged images and for the visualization of the local heartwall motion a new image analysis procedure has been developed. New parameters have been derived which allows quantification of the motion patterns and are highly sensitive to any changes in these patterns. The new procedure has been applied for heart motion analysis in healthy volunteers and in patient collectives with different heart diseases. The achieved results are summarized and discussed.

Evolution of spirituality and religiousness in chronic schizophrenia or schizo-affective disorders: A 3 years follow-up study

Purpose : Spirituality and religiousness have been shown to be highly prevalent in patients with schizophrenia. Religion can help instil a positive sense of self, decrease the impact of symptoms and provide social contacts. Religion may also be a source of suffering. In this context, this research explores whether religion remains stable over time. Methods : From an initial cohort of 115 out-patients, 80% completed the 3-years follow-up assessment. In order to study the evolution over time, a hierarchical cluster analysis using average linkage was performed on factorial scores at baseline and follow-up and their differences. A sensitivity analysis was secondarily performed to check if the outcome was influenced by other factors such as changes in mental states using mixed models. Results : Religion was stable over time for 63% patients; positive changes occurred for 20% (i.e., significant increase of religion as a resource or a transformation of negative religion to a positive one) and negative changes for 17% (i.e., decrease of religion as a resource or a transformation of positive religion to a negative one). Change in spirituality and/or religiousness was not associated with social or clinical status, but with reduced subjective quality of life and self-esteem; even after controlling for the influence of age, gender, quality of life and clinical factors at baseline. Conclusions : In this context of patients with chronic schizophrenia, religion appeared to be labile. Qualitative analyses showed that those changes expressed the struggles of patients and suggest that religious issues need to be discussed in clinical settings.

Magnetic susceptibility to identify landscape segments on a detailed scale in the region of Jaboticabal, São Paulo, Brazil

The agricultural potential is generally assessed and managed based on a one-dimensional vision of the soil profile, however, the increased appreciation of sustainable production has stimulated studies on faster and more accurate evaluation techniques and methods of the agricultural potential on detailed scales. The objective of this study was to investigate the possibility of using soil magnetic susceptibility for the identification of landscape segments on a detailed scale in the region of Jaboticabal, São Paulo State. The studied area has two slope curvatures: linear and concave, subdivided into three landscape segments: upper slope (US, concave), middle slope (MS, linear) and lower slope (LS, linear). In each of these segments, 20 points were randomly sampled from a database with 207 samples forming a regular grid installed in each landscape segment. The soil physical and chemical properties, CO2 emissions (FCO2) and magnetic susceptibility (MS) of the samples were evaluated represented by: magnetic susceptibility of air-dried fine earth (MS ADFE), magnetic susceptibility of the total sand fraction (MS TS) and magnetic susceptibility of the clay fraction (MS Cl) in the 0.00 - 0.15 m layer. The principal component analysis showed that MS is an important property that can be used to identify landscape segments, because the correlation of this property within the first principal component was high. The hierarchical cluster analysis method identified two groups based on the variables selected by principal component analysis; of the six selected variables, three were related to magnetic susceptibility. The landscape segments were differentiated similarly by the principal component analysis and by the cluster analysis using only the properties with higher discriminatory power. The cluster analysis of MS ADFE, MS TS and MS Cl allowed the formation of three groups that agree with the segment division established in the field. The grouping by cluster analysis indicated MS as a tool that could facilitate the identification of landscape segments and enable the mapping of more homogeneous areas at similar locations.

Soil functioning in a toposequence under rainforest in São Paulo, Brazil

Studies of soil-water dynamics using toposequences are essential to improve the understanding of soil-water-vegetation relationships. This study assessed the hydro-physical and morphological characteristics of soils of Atlantic Rainforest in the Parque Estadual de Carlos Botelho, state of São Paulo, Brazil. The study area of 10.24 ha (320 x 320 m) was covered by dense tropical rainforest (Atlantic Rainforest). Based on soil maps and topographic maps of the area, a representative transect of the soil in this plot was chosen and five soil trenches were opened to determine morphological properties. To evaluate the soil hydro-physical functioning, soil particle size distribution, bulk density (r), particle density (r s), soil water retention curves (SWRC), field saturated hydraulic conductivity (Ks), macroporosity (macro), and microporosity (micro) and total porosity (TP) were determined. Undisturbed samples were collected for micromorphometric image analysis, to determine pore size, shape, and connectivity. The soils in the study area were predominantly Inceptisols, and secondly Entisols and Epiaquic Haplustult. In the soil hydro-physical characterization of the selected transect, a change was observed in Ks between the surface and subsurface layers, from high/intermediate to intermediate/low permeability. This variation in soil-water dynamics was also observed in the SWRC, with higher water retention in the subsurface horizons. The soil hydro-physical behavior was influenced by the morphogenetic characteristics of the soils.

Variability of root traits in common bean genotypes at different levels of phosphorus supply and ontogenetic stages

Selection of common bean (Phaseolus vulgaris L.) cultivars with enhanced root growth would be a strategy for increasing P uptake and grain yield in tropical soils, but the strong plasticity of root traits may compromise their inclusion in breeding programs. The aim of this study was to evaluate the magnitude of the genotypic variability of root traits in common bean plants at two ontogenetic stages and two soil P levels. Twenty-four common bean genotypes, comprising the four growth habits that exist in the species and two wild genotypes, were grown in 4 kg pots at two levels of applied P (20 and 80 mg kg-1) and harvested at the stages of pod setting and early pod filling. Root area and root length were measured by digital image analysis. Significant genotype × P level and genotype × harvest interactions in analysis of variance indicate that the genotypic variation of root traits depended on soil nutrient availability and the stage at which evaluation was made. Genotypes differed for taproot mass, basal and lateral root mass, root area and root length at both P levels and growth stages; differences in specific root area and length were small. Genotypes with growth habits II (upright indeterminate) and III (prostrate indeterminate) showed better adaptation to limited P supply than genotypes of groups I (determinate) and IV (indeterminate climbing). Between the two harvests, genotypes of groups II and III increased the mass of basal and lateral roots by 40 and 50 %, respectively, whereas genotypes of groups I and IV by only 7 and 19 %. Values of the genotypic coefficient of determination, which estimates the proportion of phenotypic variance resulting from genetic effects, were higher at early pod filling than at pod setting. Correlations between shoot mass and root mass, which could indicate indirect selection of root systems via aboveground biomass, were higher at early pod filling than at pod setting. The results indicate that selection for root traits in common bean genotypes should preferentially be performed at the early pod-filling stage.

A post-processing pipeline to reconstruct the developing fetal brain using low-resolution MRI

Introduction. Development of the fetal brain surfacewith concomitant gyrification is one of the majormaturational processes of the human brain. Firstdelineated by postmortem studies or by ultrasound, MRIhas recently become a powerful tool for studying in vivothe structural correlates of brain maturation. However,the quantitative measurement of fetal brain developmentis a major challenge because of the movement of the fetusinside the amniotic cavity, the poor spatial resolution,the partial volume effect and the changing appearance ofthe developing brain. Today extensive efforts are made todeal with the âeurooepost-acquisitionâeuro reconstruction ofhigh-resolution 3D fetal volumes based on severalacquisitions with lower resolution (Rousseau, F., 2006;Jiang, S., 2007). We here propose a framework devoted tothe segmentation of the basal ganglia, the gray-whitetissue segmentation, and in turn the 3D corticalreconstruction of the fetal brain. Method. Prenatal MRimaging was performed with a 1-T system (GE MedicalSystems, Milwaukee) using single shot fast spin echo(ssFSE) sequences in fetuses aged from 29 to 32gestational weeks (slice thickness 5.4mm, in planespatial resolution 1.09mm). For each fetus, 6 axialvolumes shifted by 1 mm were acquired (about 1 min pervolume). First, each volume is manually segmented toextract fetal brain from surrounding fetal and maternaltissues. Inhomogeneity intensity correction and linearintensity normalization are then performed. A highspatial resolution image of isotropic voxel size of 1.09mm is created for each fetus as previously published byothers (Rousseau, F., 2006). B-splines are used for thescattered data interpolation (Lee, 1997). Then, basalganglia segmentation is performed on this superreconstructed volume using active contour framework witha Level Set implementation (Bach Cuadra, M., 2010). Oncebasal ganglia are removed from the image, brain tissuesegmentation is performed (Bach Cuadra, M., 2009). Theresulting white matter image is then binarized andfurther given as an input in the Freesurfer software(http://surfer.nmr.mgh.harvard.edu/) to provide accuratethree-dimensional reconstructions of the fetal brain.Results. High-resolution images of the cerebral fetalbrain, as obtained from the low-resolution acquired MRI,are presented for 4 subjects of age ranging from 29 to 32GA. An example is depicted in Figure 1. Accuracy in theautomated basal ganglia segmentation is compared withmanual segmentation using measurement of Dice similarity(DSI), with values above 0.7 considering to be a verygood agreement. In our sample we observed DSI valuesbetween 0.785 and 0.856. We further show the results ofgray-white matter segmentation overlaid on thehigh-resolution gray-scale images. The results arevisually checked for accuracy using the same principlesas commonly accepted in adult neuroimaging. Preliminary3D cortical reconstructions of the fetal brain are shownin Figure 2. Conclusion. We hereby present a completepipeline for the automated extraction of accuratethree-dimensional cortical surface of the fetal brain.These results are preliminary but promising, with theultimate goal to provide âeurooemovieâeuro of the normal gyraldevelopment. In turn, a precise knowledge of the normalfetal brain development will allow the quantification ofsubtle and early but clinically relevant deviations.Moreover, a precise understanding of the gyraldevelopment process may help to build hypotheses tounderstand the pathogenesis of several neurodevelopmentalconditions in which gyrification have been shown to bealtered (e.g. schizophrenia, autismâeuro¦). References.Rousseau, F. (2006), 'Registration-Based Approach forReconstruction of High-Resolution In Utero Fetal MR Brainimages', IEEE Transactions on Medical Imaging, vol. 13,no. 9, pp. 1072-1081. Jiang, S. (2007), 'MRI of MovingSubjects Using Multislice Snapshot Images With VolumeReconstruction (SVR): Application to Fetal, Neonatal, andAdult Brain Studies', IEEE Transactions on MedicalImaging, vol. 26, no. 7, pp. 967-980. Lee, S. (1997),'Scattered data interpolation with multilevel B-splines',IEEE Transactions on Visualization and Computer Graphics,vol. 3, no. 3, pp. 228-244. Bach Cuadra, M. (2010),'Central and Cortical Gray Mater Segmentation of MagneticResonance Images of the Fetal Brain', ISMRM Conference.Bach Cuadra, M. (2009), 'Brain tissue segmentation offetal MR images', MICCAI.

MORPHOLOGICAL AND MICROMORPHOLOGICAL CHANGES IN THE STRUCTURE OF A RHODIC HAPLUDOX AS A RESULT OF AGRICULTURAL MANAGEMENT

In evaluation of soil quality for agricultural use, soil structure is one of the most important properties, which is influenced not only by climate, biological activity, and management practices but also by mechanical and physico-chemical forces acting in the soil. The purpose of this study was to evaluate the influence of conventional agricultural management on the structure and microstructure of a Latossolo Vermelho distroférrico típico (Rhodic Hapludox) in an experimental area planted to maize. Soil morphology was described using the crop profile method by identifying the distinct structural volumes called Morphologically Homogeneous Units (MHUs). For comparison, we also described a profile in an adjacent area without agricultural use and under natural regrowth referred to as Memory. We took undisturbed samples from the main MHUs so as to form thin sections and blocks of soil for micromorphological and micromorphometrical analyses. Results from the application of the crop profile method showed the occurrence of the following structural types: loose (L), fragmented (F) and continuous (C) in both profiles analyzed. In the Memory soil profile, the fragmented structures were classified as Fptμ∆+tf and Fmt∆μ, whose micromorphology shows an enaulic-porphyric (porous) relative distribution with a great deal of biological activity as indicated by the presence of vughs and channels. Lower down, from 0.20 to 0.35 m, there was a continuous soil volume (sub-type C∆μ), with a subangular block microstructure and an enaulic-porphyric relative distribution, though in this case more compact and with aggregate coalescence and less biological activity. The micromorphometrical study of the soil of the Memory Plot showed the predominance of complex pores in NAM (15.03 %), Fmt∆μ (11.72 %), and Fptμ∆+tf (7.73 %), and rounded pores in C∆μ (8.21 %). In the soil under conventional agricultural management, we observed fragmented structures similar to the Memory Plot from 0.02 to 0.20 m, followed by a volume with a compact continuous structure (C∆μ), without visible porosity and with few roots. In the MHUs under conventional management, reduction in the packing pores (40 %) was observed, mainly in the continuous units (C). The microstructure had well-defined blocks, with the occurrence of planar pores and less evidence of biological activity. In conclusion, the morphological and micromorphological analyses of the soil profiles studied offered complementary information regarding soil structural quality, especially concerning the changes in pore types as result of mechanical stress undergone by the soil.

THREE DIMENSIONAL CHARACTERIZATION OF SOIL MACROPOROSITY BY X-RAY MICROTOMOGRAPHY

Analysis of the soil pore system represents an important way of characterizing soil structure. Properties such as the shape and number of pores can be determined through soil pore evaluations. This study presents a three-dimensional (3D) characterization of the shape and number of pores of a sub-tropical soil. To do so, a second generation X-ray microtomograph equipped with a plain type detector was employed. A voltage of 120 kV and current of 80 mA was applied to the X-ray tube. The soil samples analyzed were collected at three different depths (0-10, 10-20, and 20-30 cm). The results obtained allowed qualitative (images) and quantitative (3D) analyses of the soil structure, revealing the potential of the microtomographic technique, as well as the study of differences in soil macroporosity at different depths. Macroporosity was 5.14 % in the 0-10 cm layer, 5.10 % in the 10-20 cm layer, and 6.64 % in the 20-30 cm layer. The macroporosity of unclassified pores (UN) was 0.30 % (0-10 and 10-20 cm) and 0.40 % (20-30 cm), while equant pores (EQ) had values of 0.01 % at the three depths under analysis.