Karyotype description and evidence of multiple sex chromosome system X1X1X2X2/X1X2Y in Potamotrygon aff. motoro and P. falkneri (Chondrichthyes: Potamotrygonidae) in the upper Paraná River basin, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Temporal distribution and composition of the ichthyoplankton from Leopoldo's Inlet on the Upper Parana River floodplain (Brazil)

Temporal distribution and composition of ichthyoplankton and their relationships with some environmental variables of Leopoldo's Inlet (Ressaco do Leopoldo) on the upper Parana River floodplain, southern Brazil, were analysed. Samples were taken from February 1991 to February 1992, and 3480 larvae were collected. The highest density of larvae was observed in November 1991 (36.79 larvae/ 10 m(3)) at night (night/day ratio 7: 1). Species composition showed a predominance of Characiformes (80%). There was a high density (4.28 larvae/10 m(3)) of Bryconamericus stramineus throughout almost the whole study period, the majority of the species were more frequently collected at night. Pearson's correlation indicated a relationship between larval density, temperature, and pluviometric index. Leopoldo's Inlet, whose ichthyofauna is mainly composed of grazing and resident species. shows favourable conditions for fish reproduction and development.

Corumbataia britskii (Siluriformes: Loricariidae: Hypoptopomatinae) a new species from the upper Rio Paraná basin, Mato Grosso do Sul, Central Brazil

A new Hypoptopomatinae, Corumbataia britskii, is described based on specimens recently collected in a tributary of the upper Rio Paraná basin, Mato Grosso do Sul State, Brazil. Corumbataia britskii differs from its congeners by having a more depressed head profile (vs. a more rounded head profile in C. cuestae and C. tocantinensis), by having a more developed and conspicuous tuft of enlarged odontodes on the tip of the supraoccipital (vs. its reduced condition found in C. cuestae and C. tocantinensis), and by heaving a partially enclosed arrector fossae (vs. an almost completely enclosed arrector fossae in C. cuestae and C. tocantinensis). Copyright © 2007 Magnolia Press.

Connectivity processes and riparian vegetation of the upper Paraná River, Brazil

In fluvial systems, the relationship between a dominant variable (e.g. flood pulse) and its dependent ones (e.g. riparian vegetation) is called connectivity. This paper analyzes the connectivity elements and processes controlling riparian vegetation for a reach of the upper Paraná River (Brazil) and estimates the future changes in channel-vegetation relationship as a consequence of the managing of a large dam. The studied reach is situated 30km downstream from the Porto Primavera Dam (construction finished in 1999). Through aerial photography (1:25,000, 1996), RGB-CBERS satellite imagery and a previous field botany survey it was possible to elaborate a map with the five major morpho-vegetation units: 1) Tree-dominated natural levee, 2) Shrubby upper floodplain, 3) Shrub-herbaceous mid floodplain, 4) Grass-herbaceous lower floodplain and 5) Shrub-herbaceous flood runoff channel units. By use of a detailed topographic survey and statistical tools each morpho-vegetation type was analyzed according to its connectivity parameters (frequency, recurrence, permanence, seasonality, potamophase, limnophase and FCQ index) in the pre- and post-dam closure periods of the historical series. Data showed that most of the morpho-vegetation units were predicted to present changes in connectivity parameters values after dam closing and the new regime could affect, in different intensity, the river ecology and particularly the riparian vegetation. The methods used in this study can be useful for dam impact studies in other South American tropical rivers. © 2012 Elsevier Ltd.

A New Species of Hypostomus (Siluriformes: Loricariidae) from the Upper Rio Paraguay Basin, Brazil

A new small Loricariidae, Hypostomus careopinnatus, is described from the Rio Taquari drainage, upper Rio Paraguay basin, Mato Grosso, Brazil. The new species can be easily distinguished from all congeners, except Hypostomus kids, by the absence of adipose fin. Hypostomus careopinnatus is distinguished from H. levis mainly by the presence of slender bifid teeth, with mesial cusp large and rounded, and lateral cusp small and pointed (vs. spoon-shaped teeth). The new species described herein completely lacks the adipose fin and also lacks the median pre-adipose plates in almost all specimens examined. The absence of adipose fin is probably an independent acquisition for Hypostomus careopinnatus and Hypostomus levis.

Velocity structure and seismic anisotropy in the crust and upper mantle from Receiver Function analysis: three case studies in Italy

The research for this PhD project consisted in the application of the RFs analysis technique to different data-sets of teleseismic events recorded at temporary and permanent stations located in three distinct study regions: Colli Albani area, Northern Apennines and Southern Apennines. We found some velocity models to interpret the structures in these regions, which possess very different geologic and tectonics characteristics and therefore offer interesting case study to face. In the Colli Albani some of the features evidenced in the RFs are shared by all the analyzed stations: the Moho is almost flat and is located at about 23 km depth, and the presence of a relatively shallow limestone layer is a stable feature; contrariwise there are features which vary from station to station, indicating local complexities. Three seismic stations, close to the central part of the former volcanic edifice, display relevant anisotropic signatures­­­ with symmetry axes consistent with the emplacement of the magmatic chamber. Two further anisotropic layers are present at greater depth, in the lower crust and the upper mantle, respectively, with symmetry axes directions related to the evolution of the volcano complex. In Northern Apennines we defined the isotropic structure of the area, finding the depth of the Tyrrhenian (almost 25 km and flat) and Adriatic (40 km and dipping underneath the Apennines crests) Mohos. We determined a zone in which the two Mohos overlap, and identified an anisotropic body in between, involved in the subduction and going down with the Adiratic Moho. We interpreted the downgoing anisotropic layer as generated by post-subduction delamination of the top-slab layer, probably made of metamorphosed crustal rocks caught in the subduction channel and buoyantly rising toward the surface. In the Southern Apennines, we found the Moho depth for 16 seismic stations, and highlighted the presence of an anisotropic layer underneath each station, at about 15-20 km below the whole study area. The moho displays a dome-like geometry, as it is shallow (29 km) in the central part of the study area, whereas it deepens peripherally (down to 45 km); the symmetry axes of anisotropic layer, interpreted as a layer separating the upper and the lower crust, show a moho-related pattern, indicated by the foliation of the layer which is parallel to the Moho trend. Moreover, due to the exceptional seismic event occurred on April 6th next to L’Aquila town, we determined the Vs model for two station located next to the epicenter. An extremely high velocity body is found underneath AQU station at 4-10 km depth, reaching Vs of about 4 km/s, while this body is lacking underneath FAGN station. We compared the presence of this body with other recent works and found an anti-correlation between the high Vs body, the max slip patches and earthquakes distribution. The nature of this body is speculative since such high velocities are consistent with deep crust or upper mantle, but can be interpreted as a as high strength barrier of which the high Vs is a typical connotation.

Two-step postmortem angiography with a modified heart-lung machine: preliminary results

OBJECTIVE: The purpose of this study was to adapt and improve a minimally invasive two-step postmortem angiographic technique for use on human cadavers. Detailed mapping of the entire vascular system is almost impossible with conventional autopsy tools. The technique described should be valuable in the diagnosis of vascular abnormalities. MATERIALS AND METHODS: Postmortem perfusion with an oily liquid is established with a circulation machine. An oily contrast agent is introduced as a bolus injection, and radiographic imaging is performed. In this pilot study, the upper or lower extremities of four human cadavers were perfused. In two cases, the vascular system of a lower extremity was visualized with anterograde perfusion of the arteries. In the other two cases, in which the suspected cause of death was drug intoxication, the veins of an upper extremity were visualized with retrograde perfusion of the venous system. RESULTS: In each case, the vascular system was visualized up to the level of the small supplying and draining vessels. In three of the four cases, vascular abnormalities were found. In one instance, a venous injection mark engendered by the self-administration of drugs was rendered visible by exudation of the contrast agent. In the other two cases, occlusion of the arteries and veins was apparent. CONCLUSION: The method described is readily applicable to human cadavers. After establishment of postmortem perfusion with paraffin oil and injection of the oily contrast agent, the vascular system can be investigated in detail and vascular abnormalities rendered visible.

Stricture of the Afferent Isoperistaltic Tubular Segment: A Late and Rare Cause of Bilateral Dilation of the Upper Urinary Tract After Ileal Bladder Substitution

OBJECTIVE To evaluate the etiology and treatment of bilateral hydronephrosis not responding to bladder substitute drainage after ileal bladder substitution using an afferent isoperistaltic tubular segment. MATERIALS AND METHODS A retrospective analysis was performed of a consecutive series of 739 patients who had undergone bladder substitution from April 1985 to August 2012. RESULTS Of the 739 ileal bladder substitute patients, 10 (1.4%) developed bilateral hydronephrosis unresponsive to complete bladder substitute drainage. The etiology was stenosis of the afferent isoperistaltic tubular segment. The median interval to presentation was 131 months (range 45-192). The incidence of afferent tubular segment stenosis was significantly higher in the 61 ileal bladder substitute patients with recurrent urinary tract infection (9 [15%]) than in the 678 without recurrent urinary tract infection (1 [0.15%]; P <.001). Urine cultures revealed mixed infections (34%), Escherichia coli (18%), Staphylococcus aureus (13%), enterococci (11%), Candida (8%), Klebsiella (8%), and others (8%). Seven patients underwent 10 endourologic interventions, only 1 of which was successful (10%). After failed endourologic treatment, 7 open surgical revisions with resection of the stricture were performed, with all 7 (100%) successful. CONCLUSION Bilateral dilation of the upper urinary tract after ileal orthotopic bladder substitution unresponsive to complete bladder substitute drainage is likely to be caused by stenosis of the afferent isoperistaltic tubular segment. The stenosis occurs almost exclusively in patients with long-lasting, recurrent urinary tract infection and can develop many years after the ileal bladder substitution. Minimally invasive endourologic treatment is usually unsuccessful; however, open surgical revision offers excellent results.

Classifying and mapping the urban transition in Vietnam

The urban transition almost always involves wrenching social adjustment as small agricultural communities are forced to adjust rapidly to industrial ways of life. Large-scale in-migration of young people, usually from poor regions, creates enormous demand and expectations for community and social services. One immediate problem planners face in approaching this challenge is how to define, differentiate, and map what is rural, urban, and transitional (i.e., peri-urban). This project established an urban classification for Vietnam by using national census and remote sensing data to identify and map the smallest administrative units for which data are collected as rural, peri-urban, urban, or urban core. We used both natural and human factors in the quantitative model: income from agriculture, land under agriculture and forests, houses with modern sanitation, and the Normalized Difference Vegetation Index. Model results suggest that in 2006, 71% of Vietnam's 10,891 communes were rural, 18% peri-urban, 3% urban, and 4% urban core. Of the communes our model classified as peri-urban, 61% were classified by the Vietnamese government as rural. More than 7% of Vietnam's land area can be classified as peri-urban and approximately 13% of its population (more than 11 million people) lives in peri-urban areas. We identified and mapped three types of peri-urban places: communes in the periphery of large towns and cities; communes along highways; and communes associated with provincial administration or home to industrial, energy, or natural resources projects (e.g., mining). We validated this classification based on ground observations, analyses of multi-temporal night-time lights data, and an examination of road networks. The model provides a method for rapidly assessing the rural–urban nature of places to assist planners in identifying rural areas undergoing rapid change with accompanying needs for investments in building, sanitation, road infrastructure, and government institutions.

Emergence of acquired HIV-1 drug resistance has almost been stopped in Switzerland - a 15 year prospective cohort analysis

BACKGROUND  Drug resistance is a major barrier to successful antiretroviral treatment (ART). Therefore, it is important to monitor time trends at a population level. METHODS  We included 11,084 ART-experienced patients from the Swiss HIV Cohort Study (SHCS) between 1999 and 2013. The SHCS is highly representative and includes 72% of patients receiving ART in Switzerland. Drug resistance was defined as the presence of at least one major mutation in a genotypic resistance test. To estimate the prevalence of drug resistance, data for patients with no resistance test was imputed based on patient's risk of harboring drug resistant viruses. RESULTS  The emergence of new drug resistance mutations declined dramatically from 401 to 23 patients between 1999 and 2013. The upper estimated prevalence limit of drug resistance among ART-experienced patients decreased from 57.0% in 1999 to 37.1% in 2013. The prevalence of three-class resistance decreased from 9.0% to 4.4% and was always <0.4% for patients who initiated ART after 2006. Most patients actively participating in the SHCS in 2013 with drug resistant viruses initiated ART before 1999 (59.8%). Nevertheless, in 2013, 94.5% of patients who initiated ART before 1999 had good remaining treatment options based on Stanford algorithm. CONCLUSION  HIV-1 drug resistance among ART-experienced patients in Switzerland is a well-controlled relic from the pre-combination ART era. Emergence of drug resistance can be virtually stopped with new potent therapies and close monitoring.

Gravity mapping in the southern Weddell Sea region, Antarctica (Scale 1:2 000 000)

New maps of free-air and the Bouguer gravity anomalies on the Weddell Sea sector (70-81° S, 6-75° W) of Antarctica are presented. These maps are based on the first computer compilation of available gravity data collected by ''Sevmorgeologia'' in 1976-89 in the southern Weddell Sea and adjacent coasts of western Dronning Maud Land (WDML) and Coats Land. The accomplished gravity studies comprise airborne observations with a line spacing of about 20 km and conventional measurements at over-the-ice points, which were spaced at 10-30 km and supplemented by seismic soundings. Hence, anomalies on the maps represent mainly large-scale and deep crustal features. The dominant feature in free-air gravity map is a large dipolar gravity anomaly stretching along the continental margin. Following the major grain of seabed morphology this shelf-edge/slope anomaly (SESA) is clearly divided into three segments characterized by diverse anomaly amplitudes, wavelengths and trends. They are associated with continental margins of different geotectonic provinces of Antarctica surrounding the Weddell Sea. Apparent distinctions in the SESA signatures are interpreted as the gravity expression of tectonic, deep crustal structure segmentation of the continental margin. The prominent gravity highs (100-140 mGal) of the shelf edge anomaly mapped along WDML are assumed to represent high-density mantle injections intruded into the middle/lower crust during initial rifting of continental breakup. Enlarged wavelengths and diminished amplitudes of the gravity anomaly westwards, along the Weddell Sea embayment (WSE) margin, reflect a widening of the continental slope and a significant increase in thickness of underlying sediment strata. Low amplitude, negative free-air anomalies in the Filchner-Ronne Ice Shelves (FRIS) contrast sharply with the dominating positive anomalies offshore. This indicates a greater sedimentary thickness of the basin in this area. Crustal response to the enlarged sediment load is impressed in mostly positive features of the Bouguer gravity field observed here. Two pronounced positive Bouguer anomalies of 50-70 mGal and an average widths of 200 km dominate the Weddell Sea embayment margins towards the Antarctic Peninsula and the East Antarctic craton. They correlate well with very deep seabed troughs (> 1000 m below sea level). The gravity highs are most likely caused by a shallow upper mantle underneath graben-rift structures evolved at the margins of the WSE basin. A regional zone (> 100 km in width) of the prominent Bouguer and free-air negative anomalies (-40 to -60 mGal) adjacent Coats Land to the north of the ice shelf edge may indicate the presence of the thick old cratonic crust far offshore beneath the Weddell Sea Embayment.

Two pollen profiles of the middle Pleistocene at Samerberg/Upper Bavaria, Germany

Previous pollen analytical studies on sediments from the pleistocene lake basin at Samerberg, situated on the northern edge of the Bavarian Alps (47°45' N, 12°12' E, 607 m a.s.l.) had been performed on samples taken from cores and exposures close to the southern shore of the former lake. After geoelectric and refraction-seismic measurements had shown that the lake basin had been much deeper in its northern part, another core was taken where maximum depth could be expected. The corer penetrated three moraines, two of them lying above pollen-bearing sediments, and one below them, and reached the hard rock (Kössener Kalk) at a depth of 93 m. Two forest phases could be identified by pollen analysis. The pollen record begins abruptly in a forest phase at the end of a spruce-dominated period when fir started to spread (DA 1, DA = pollen zone). Following this, Abies (fir) was the main tree species at Samerberg, Picea being second, and deciduous trees were almost non-existent. First box (Buxus) was of major importance in the fir forests (DA 2), but later on beech (Fagus) and wing-nut (Pterocarya) spread (DA 3). Finally this forest gave way to a spruce forest with pine (DA 4). The beginning and the end of this interglacial cycle are not recorded. Its vegetational development is different from the eemian one known from earlier studies at Samerberg. It is characterized by the occurrence of Abies together with Buxus, Pterocarya and Fagus. A similar association of woody species is known only from the Holsteinian age deposits in an area ranging from England to Poland, though at no other place these species were such important constituents of the vegetation as at Samerberg. Therefore zone 1 to 4 are attributed to the Holsteinian interglacial period. The younger forest phase, separated from the interglacial by a stadial with open vegetation (DA 5), seems to be completely represented, though its sediments are disturbed, apparently by sliding which caused repetition of same-age-sediments in the core (DA 7a, b, c) The vegetational development is simple. A juniper phase (DA 6) was followed by reforestation with spruce, accompanied by some fir (DA 7, 9). Finally pine became the dominant species (DA 9). The simple vegetational development of this younger forest phase does not allow a safe correlation with one of the known pre-eemian interstadials, but for stratigraphical reasons it can be related best to the Dömnitz-interglacial, which among others is also known as Wacken- or Holstein-II-interglacial. Possibly another phase of reforestation is indicated at the end of the following stadial (DA 10). But due to an erosional unconformity nothing than the rise of the juniper curve can be stated. It was only after this sequence of forest phases and periods with open vegetation that glaciers reached the Samerberg area again.

A geometric characterization of the upper bound for the span of the jones polynomial

Let D be a link diagram with n crossings, sA and sB be its extreme states and |sAD| (respectively, |sBD|) be the number of simple closed curves that appear when smoothing D according to sA (respectively, sB). We give a general formula for the sum |sAD| + |sBD| for a k-almost alternating diagram D, for any k, characterizing this sum as the number of faces in an appropriate triangulation of an appropriate surface with boundary. When D is dealternator connected, the triangulation is especially simple, yielding |sAD| + |sBD| = n + 2 - 2k. This gives a simple geometric proof of the upper bound of the span of the Jones polynomial for dealternator connected diagrams, a result first obtained by Zhu [On Kauffman brackets, J. Knot Theory Ramifications6(1) (1997) 125–148.]. Another upper bound of the span of the Jones polynomial for dealternator connected and dealternator reduced diagrams, discovered historically first by Adams et al. [Almost alternating links, Topology Appl.46(2) (1992) 151–165.], is obtained as a corollary. As a new application, we prove that the Turaev genus is equal to the number k of dealternator crossings for any dealternator connected diagram

Urban trees and atmospheric pollutants in big cities: Effects in Madrid

Una investigación sobre la mejora de la contaminación del aire (CA) por medio de arbolado urbano se realizó en Madrid, una ciudad con casi 4 M de habitantes, 2,8 M de vehículos y casi 3 M de árboles de mantenimiento público. La mayoría de los árboles estaban en dos bosques periurbanos. Los 650.000 restantes era pies de alineación y parques. Los taxones estudiados fueron Platanus orientalis (97.205 árboles), Ulmus sp. (70.557), Pinus pinea (49.038), Aesculus hippocastanum (22.266), Cedrus sp. (13.678) y Quercus ilex (1.650), de calles y parques. Muestras foliares se analizaron en diferentes épocas del año, así como datos de contaminación por PM10 de 28 estaciones de medición de la contaminación durante 30 años, y también la intensidad del tráfico (IMD) en 2.660 calles. La acumulación de metales pesados (MP) sobre hojas y dentro de estas se estimó en relación con la CA y del suelo y la IMD del tráfico. La concentración media de Ba, Cd, Cr, Cu, Mn, Ni, Pb y Zn en suelo (materia seca) alcanzó: 489,5, 0,7, 49,4, 60,9, 460,9, 12,8, 155,9 y 190,3 mg kg-1 respectivamente. Los árboles urbanos, particularmente coníferas (debido a la mayor CA en invierno) contribuyen significativamente a mejorar la CA sobre todo en calles con alta IMD. La capacidad de las seis sp. para capturar partículas de polvo en su superficies foliares está relacionada con la IMD del tráfico y se estimó en 16,8 kg/año de MP tóxicos. Pb y Zn resultaron ser buenos marcadores antrópicos en la ciudad en relación con el tráfico, que fue la principal fuente de contaminación en los árboles y suelos de Madrid. Las especies de árboles variaron en función de su capacidad para capturar partículas (dependiendo de las propiedades de sus superficies foliares) y acumular los MP absorbidos de los suelos. Las concentraciones foliares de Pb y Zn estuvieron por encima de los límites establecidos en diferentes sitios de la ciudad. La microlocalización de Zn mediante microscópico mostró la translocación al xilema y floema. Se detectaron puntos de contaminación puntual de Cu and Cr en antiguos polígonos industriales y la distribución espacial de los MP en los suelos de Madrid mostró que en incluso en zonas interiores del El Retiro había ciertos niveles elevados de [Pb] en suelo, tal vez por el emplazamiento la Real Fábrica de Porcelana en la misma zona hace 200 años. Distintas áreas del centro de la ciudad también alcanzaron niveles altos de [Pb] en suelo. Según los resultados, el empleo de una combinación de Pinus pinea con un estrato intermedio de Ulmus sp. y Cedrus sp. puede ser la mejor recomendación como filtro verde eficiente. El efecto del ozono (O3) sobre el arbolado en Madrid fue también objeto de este estudio. A pesar de la reducción de precursores aplicada en muchos países industrializados, O3 sigue siendo la principal causa de CA en el hemisferio norte, con el aumento de [O3] de fondo. Las mayores [O3] se alcanzaron en regiones mediterráneas, donde el efecto sobre la vegetación natural es compensado por el xeromorfismo y la baja conductancia estomática en respuesta los episodios de sequía estival característicos de este clima. Durante una campaña de monitoreo, se identificaron daños abióticos en hojas de encina parecidos a los de O3 que estaban plantadas en una franja de césped con riego del centro de Madrid. Dada la poca evidencia disponible de los síntomas de O3 en frondosas perennifolias, se hizo un estudio que trató de 1) confirman el diagnóstico de daño de O3, 2) investigar el grado de los síntomas en encinas y 3) analizar los factores ambientales que contribuyeron a los daños por O3, en particular en lo relacionado con el riego. Se analizaron los marcadores macro y micromorfológicos de estrés por O3, utilizando las mencionadas encinas a modo de parcela experimental. Los síntomas consistieron en punteado intercostal del haz, que aumentó con la edad. Además de un punteado subyacente, donde las células superiores del mesófilo mostraron reacciones características de daños por O3. Las células próximas a las zonas dañadas, presentaron marcadores adicionales de estrés oxidativo. Estos marcadores morfológicos y micromorfológicos de estrés por O3 fueron similares a otras frondosas caducifolias con daños por O3. Sin embargo, en nuestro caso el punteado fue evidente con AOT40 de 21 ppm•h, asociada a riego. Análisis posteriores mostraron que los árboles con riego aumentaron su conductancia estomática, con aumento de senescencia, manteniéndose sin cambios sus características xeromórficas foliares. Estos hallazgos ponen de relieve el papel primordial de la disponibilidad de agua frente a las características xeromórficas a la hora de manifestarse los síntomas en las células por daños de O3 en encina. ABSTRACT Research about air pollution mitigation by urban trees was conducted in Madrid (Spain), a southern European city with almost 4 M inhabitants, 2.8 M daily vehicles and 3 M trees under public maintenance. Most trees were located in two urban forests, while 650'000 trees along urban streets and in parks. The urban taxa included Platanus orientalis (97'205 trees), Ulmus sp. (70’557), Pinus pinea (49'038), Aesculus hippocastanum (22’266), Cedrus sp. (13'678 and Quercus ilex (1'650) along streets and parks. Leave samples were analysed sequentially in different seasons, PM10 data from 28 air monitoring stations during 30 years and traffic density estimated from 2’660 streets. Heavy metal (HM) accumulation on the leaf surface and within leaves was estimated per tree related to air and soil pollution, and traffic intensity. Mean concentration of Ba, Cd, Cr, Cu, Mn, Ni, Pb and Zn in topsoil samples (dry mass) amounted in Madrid: 489.5, 0.7, 49.4, 60.9, 460.9, 12.8, 155.9 and 190.3 mg kg-1 respectively. Urban trees, particularly conifers (due to higher pollution in winter) contributed significantly to alleviate air pollution especially near to high ADT roads. The capacity of the six urban street trees species to capture air-born dust on the foliage surface as related to traffic intensity was estimated to 16.8 kg of noxious metals from exhausts per year. Pb and Zn pointed to be tracers of anthropic activity in the city with vehicle traffic as the main source of diffuse pollution on trees and soils. Tree species differed by their capacity to capture air-borne dust (by different leaf surface properties) and to allocate HM from soils. Pb and Zn concentrations in the foliage were above limits in different urban sites and microscopic Zn revelation showed translocation in xylem and phloem tissue. Punctual contamination in soils by Cu and Cr was identified in former industrial areas and spatial trace element mapping showed for central Retiro Park certain high values of [Pb] in soils even related to a Royal pottery 200 years ago. Different areas in the city centre also reached high levels [Pb] in soils. According to the results, a combination of Pinus pinea with understorey Ulmus sp. and Cedrus sp. layers can be recommended for the best air filter efficiency. The effects of ozone (O3) on trees in different areas of Madrid were also part of this study. Despite abatement programs of precursors implemented in many industrialized countries, ozone remained the main air pollutant throughout the northern hemisphere with background [O3] increasing. Some of the highest ozone concentrations were measured in regions with a Mediterranean climate but the effect on the natural vegetation is alleviated by low stomatal uptake and frequent leaf xeromorphy in response to summer drought episodes characteristic of this climate. During a bioindication survey, abiotic O3-like injury was identified in foliage. Trees were growing on an irrigated lawn strip in the centre of Madrid. Given the little structural evidence available for O3 symptoms in broadleaved evergreen species, a study was undertaken in 2007 with the following objectives 1) confirm the diagnosis, 2) investigate the extent of symptoms in holm oaks growing in Madrid and 3) analyse the environmental factors contributing to O3 injury, particularly, the site water supply. Therefore, macro- and micromorphological markers of O3 stress were analysed, using the aforementioned lawn strip as an intensive study site. Symptoms consisted of adaxial and intercostal stippling increasing with leaf age. Underlying stippling, cells in the upper mesophyll showed HR-like reactions typical of ozone stress. The surrounding cells showed further oxidative stress markers. These morphological and micromorphological markers of ozone stress were similar to those recorded in deciduous broadleaved species. However, stippling became obvious already at an AOT40 of 21 ppm•h and was primarily found at irrigated sites. Subsequent analyses showed that irrigated trees had their stomatal conductance increased and leaf life-span reduced whereas their leaf xeromorphy remained unchanged. These findings suggest a central role of water availability versus leaf xeromorphy for ozone symptom expression by cell injury in holm oak.

Multiple-complete-digest restriction fragment mapping: Generating sequence-ready maps for large-scale DNA sequencing

Multiple-complete-digest mapping is a DNA mapping technique based on complete-restriction-digest fingerprints of a set of clones that provides highly redundant coverage of the mapping target. The maps assembled from these fingerprints order both the clones and the restriction fragments. Maps are coordinated across three enzymes in the examples presented. Starting with yeast artificial chromosome contigs from the 7q31.3 and 7p14 regions of the human genome, we have produced cosmid-based maps spanning more than one million base pairs. Each yeast artificial chromosome is first subcloned into cosmids at a redundancy of ×15–30. Complete-digest fragments are electrophoresed on agarose gels, poststained, and imaged on a fluorescent scanner. Aberrant clones that are not representative of the underlying genome are rejected in the map construction process. Almost every restriction fragment is ordered, allowing selection of minimal tiling paths with clone-to-clone overlaps of only a few thousand base pairs. These maps demonstrate the practicality of applying the experimental and software-based steps in multiple-complete-digest mapping to a target of significant size and complexity. We present evidence that the maps are sufficiently accurate to validate both the clones selected for sequencing and the sequence assemblies obtained once these clones have been sequenced by a “shotgun” method.