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1. Non-matching predictions from different models simulating the effects of elevated atmospheric CO2 on the Amazon forest’s functional diversity

Author

Blanco, C.C., Rius, B.F., Darela-Filho, J.P., Cardeli, B., Aleixo, I., Scheiter, S., Langan, L., Joshi, J., Hofhansl, F., Singh, S., De Paula, M.D., Hickler, T., Ongole, S., Higgins, S., Fleischer, K., Rammig, A., Lichstein, J., Lapola, D.M., Blanco, C.C., Rius, B.F., Darela-Filho, J.P., Cardeli, B., Aleixo, I., Scheiter, S., Langan, L., Joshi, J., Hofhansl, F., Singh, S., De Paula, M.D., Hickler, T., Ongole, S., Higgins, S., Fleischer, K., Rammig, A., Lichstein, J., and Lapola, D.M.

Abstract

The continuous rising of atmospheric carbon dioxide (CO2) concentration is undoubtedly affecting the resilience of tropical forests worldwide. However, the magnitude of such effects is poorly known, limiting our capacity to assess the vulnerability of tropical forests and to improve their representation by models. Functional diversity (FD) is an important component of biodiversity enhancing ecosystem resilience, as high FD can provide higher response diversity and capacity to buffer against climate change. How FD is represented by different Dynamic Global Vegetation Models (DGVMs) may affect how such models predict the impacts of environmental changes on hyperdiverse ecosystems. We compared simulations of five trait-based DGVMs (i.e., with flexible, variable traits) constrained with data from the Amazon rainforest in the scope of the AmazonFACE project. Simulations were conducted considering initial high or low diversity scenarios under ambient and elevated CO2 (400 ppm and 600 ppm, respectively). We searched for correspondence between the functional identity of simulated plant strategies and their ecophysiological performances under elevated CO2. As models take different approaches to simulating functional trait distributions and they differ in their structure and in the trade-offs implemented, we found important intermodel differences in simulated results. Nevertheless, we took advantage of these differences in order to assess the most likely scenarios in terms of functional composition under elevated CO2, as well as to give feedback for better harmonization of model inputs and outputs and future model improvements. In the face of the pessimistic scenarios that project a continuous increase in CO2 levels, resolving the divergent responses among model predictions is critical, given the global importance of the Amazon rainforest's biodiversity and climate regulation, as well as the approximately 30 million people that directly or indirectly depend on the forest for

Published
2024
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5. Competition for light can drive adverse species-composition shifts in the Amazon Forest under elevated CO2

Author

Joshi, J., Hofhansl, F., Singh, S., Stocker, B., Brännström, Å., Franklin, O., Blanco, C.C., Aleixo, I., Lapola, D.M., Prentice, I.C., and Dieckmann, U.

Abstract

The resilience of biodiverse forests to climate change depends on an interplay of adaptive processes operating at multiple temporal and organizational scales. These include short-term acclimation of physiological processes like photosynthesis and respiration, mid-term changes in forest structure due to competition, and long-term changes in community composition arising from competitive exclusion and genetic trait evolution. To investigate the roles of diversity and adaptation for forest resilience, we present Plant-FATE, a parsimonious eco-evolutionary vegetation model. Tested with data from a hyperdiverse Amazonian terra-firme forest, our model accurately predicts multiple emergent ecosystem properties characterizing forest structure and function. Under elevated CO2 conditions, we predict an increase in productivity, leaf area, and aboveground biomass, with the magnitude of this increase declining in nutrient-deprived soils if trees allocate more carbon to the rhizosphere to overcome nutrient limitation. Furthermore, increased aboveground productivity leads to greater competition for light and drives a shift in community composition towards fast-growing but short-lived species characterized by lower wood densities. Such a transition reduces the carbon residence time of woody biomass, dampening carbon-sink strength and potentially rendering the Amazon Forest more vulnerable to future climatic extreme events.

Published
2023

7. Predicting the adaptive responses of biodiverse plant communities using functional trait evolution

Author

Joshi, J., Hofhansl, F., Singh, S., Stocker, B., Vignal, T., Brännström, Å., Franklin, O., Blanco, C., Aleixo, I., Lapola, D., Prentice, I., Dieckmann, U., Joshi, J., Hofhansl, F., Singh, S., Stocker, B., Vignal, T., Brännström, Å., Franklin, O., Blanco, C., Aleixo, I., Lapola, D., Prentice, I., and Dieckmann, U.

Abstract

Climate change consists of synergistic changes in a wide range of environmental conditions, characterized by elevated CO2, higher mean temperatures, and higher climate variability. While elevated CO2 concentrations may potentially increase the productivity of some ecosystems, it has been argued that nutrient limitation, increased respiration, and increased mortality may dampen or even negate these productivity gains. The capacity of global forests to adjust to such synergistic environmental changes depends on their functional diversity and the ecosystem’s adaptive capacity. The Plant-FATE eco-evolutionary model describes vegetation responses to altered environmental conditions, including CO2 concentrations, temperature, and water limitation. It represents functional diversity by modelling species as points in trait space and incorporates ecosystem adaptations at three levels: 1) to model acclimation of plastic traits of individual plants, we leverage the power of eco-evolutionary optimality principles, 2) to model shifts in species composition via demographic changes and species immigration, we implement a trait-size-structured demographic vegetation model, and 3) to model the long-term genetic evolution of species, we have developed new evolutionary theory for trait-size-structured communities. First, we show that with just a few calibrated parameters, the Plant-FATE model accurately predicts the fluxes of CO2 and water, size distributions, and trait distributions for a tropical wet site in the Amazon Forest. Second, we show that under elevated CO2 our model predictions are broadly consistent with observations, namely: an increase in leaf area, productivity and biomass, and a decrease in stomatal conductance and photosynthetic capacity. Third, we show that CO2 and nutrient fertilization both drive changes in community composition towards fast life-histories, and that competition drives the system in a direction opposite to what is optimal for individual plants. Our

Published
2023

8. Roles of diversity and adaptation in the eco-evolutionary responses of biodiverse plant communities to climate change

Author

Joshi, J., Hofhansl, F., Singh, S., Stocker, B., Brännström, Å., Vignal, T., Casagrande Blanco, C., Aleixo, I., Lapola, D., Prentice, I.C., Dieckmann, U., Joshi, J., Hofhansl, F., Singh, S., Stocker, B., Brännström, Å., Vignal, T., Casagrande Blanco, C., Aleixo, I., Lapola, D., Prentice, I.C., and Dieckmann, U.

Abstract

Climate change is projected to cause not only higher mean temperatures but also higher climate variability. Although elevated CO2 concentrations can potentially increase the productivity of some ecosystems, higher temperatures and more frequent droughts may lead to increased respiration and mortality, possibly negating these productivity gains. The capacity of global forests to adjust to climate change depends on their functional diversity and the ecosystem’s adaptive capacity. The Plant-FATE eco-evolutionary model describes vegetation responses to altered environmental conditions, including CO2 concentrations, temperatures, and droughts. It represents functional diversity by modelling species as points in trait space and incorporates ecosystem adaptations at three levels: 1) to model acclimation of plastic traits of individual plants, we leverage the power of eco-evolutionary optimality principles, 2) to model shifts in species composition via demographic changes and species immigration, we implement a trait-size-structured demographic vegetation model, and 3) to model the long-term genetic evolution of species, we have developed new evolutionary theory for trait-size-structured communities. First, we show that with just a few calibrated parameters, the Plant-FATE model accurately predicts the fluxes of CO2 and water, size distributions, and trait distributions for a tropical wet site in the Amazon Forest. Second, we show that under elevated CO2 conditions and in the absence of nutrient limitation, our model predictions are broadly consistent with observations, namely: an increase in leaf area, productivity and biomass, and a decrease in stomatal conductance and photosynthetic capacity. Third, we simulate the calibrated model with hypothetical future drought regimes to investigate three key features of ecosystem responses: 1) the change in species composition and ecosystem functioning in response to altered conditions, 2) the timescales of ecosystem response to new

Published
2023

16. TRY plant trait database - enhanced coverage and open access

Author

Kattge, J, Bönisch, G, Díaz, S, Lavorel, S, Prentice, IC, Leadley, P, Tautenhahn, A, Werner, GDA, Aakala, T, Abedi, M, Acosta, ATR, Adamidis, GC, K, Adamson., Aiba, M, Albert, CH, Alcántara, JM, Alcázar, C, Aleixo, I, Ali, H, Amiaud, B, Ammer, C, Amoroso, MM, Anand, M, Anderson, C, Anten, N, Antos, J, Apgaua, DMG, Ashman, TL, Asmara, DH, Asner, GP, Aspinwall, M, Atkin, O, Aubin, I, Baastrup-Spohr, L, Bahalkeh, K, Bahn, M, Baker, T, Baker, WJ, Bakker, JP, Baldocchi, D, Baltzer, J, Banerjee, A, Baranger, A, Barlow, J, Barneche, DR, Baruch, Z, Bastianelli, D, Battles, J, Salguero-Gomez, R, and Terrestrial Ecology (TE)

Subjects
Access to Information, Ecology, Plan_S-Compliant-TA, international, food and beverages, Biodiversity, Plants, Ecosystem
Abstract

Plant traits-the morphological, anatomical, physiological, biochemical and phenological characteristics of plants-determine how plants respond to environmental factors, affect other trophic levels, and influence ecosystem properties and their benefits and detriments to people. Plant trait data thus represent the basis for a vast area of research spanning from evolutionary biology, community and functional ecology, to biodiversity conservation, ecosystem and landscape management, restoration, biogeography and earth system modelling. Since its foundation in 2007, the TRY database of plant traits has grown continuously. It now provides unprecedented data coverage under an open access data policy and is the main plant trait database used by the research community worldwide. Increasingly, the TRY database also supports new frontiers of trait-based plant research, including the identification of data gaps and the subsequent mobilization or measurement of new data. To support this development, in this article we evaluate the extent of the trait data compiled in TRY and analyse emerging patterns of data coverage and representativeness. Best species coverage is achieved for categorical traits-almost complete coverage for 'plant growth form'. However, most traits relevant for ecology and vegetation modelling are characterized by continuous intraspecific variation and trait-environmental relationships. These traits have to be measured on individual plants in their respective environment. Despite unprecedented data coverage, we observe a humbling lack of completeness and representativeness of these continuous traits in many aspects. We, therefore, conclude that reducing data gaps and biases in the TRY database remains a key challenge and requires a coordinated approach to data mobilization and trait measurements. This can only be achieved in collaboration with other initiatives.

Published
2020

30. TRY plant trait database enhanced coverage and open access

Author

Kattge, J., Bönisch, G., Díaz, S., Lavorel, S., Prentice, I.C., Leadley, P., Tautenhahn, S., Werner, G.D.A., Aakala, T., Abedi, M., Acosta, A.T.R., Adamidis, G.C., Adamson, K., Aiba, M., Albert, C.H., Alcántara, J.M., Alcázar, C, C., Aleixo, I., Ali, H., Amiaud, B., Ammer, C., Amoroso, M.M., Anand, M., Anderson, C., Anten, N., Antos, J., Apgaua, D.M.G., Ashman, T.-L., Asmara, D.H., Asner, G.P., Aspinwall, M., Atkin, O., Aubin, I., Baastrup-Spohr, L., Bahalkeh, K., Bahn, M., Baker, T., Baker, W.J., Bakker, J.P., Baldocchi, D., Baltzer, J., Banerjee, A., Baranger, A., Barlow, J., Barneche, D.R., Baruch, Z., Bastianelli, D., Battles, J., Bauerle, W., Bauters, M., Bazzato, E., Beckmann, M., Beeckman, H., Beierkuhnlein, C., Bekker, R., Belfry, G., Belluau, M., Beloiu, M., Benavides, R., Benomar, L., Berdugo-Lattke, M.L., Berenguer, E., Bergamin, R., Bergmann, J., Bergmann, Carlucci, M., Berner, L., Bernhardt-Römermann, M., Bigler, C., Bjorkman, A.D., Blackman, C., Blanco, C., Blonder, B., Blumenthal, D., Bocanegra-González, K.T., Boeckx, P., Bohlman, S., Böhning-Gaese, K., Boisvert-Marsh, L., Bond, W., Bond-Lamberty, B., Boom, A., Boonman, C.C.F., Bordin, K., Boughton, E.H., Boukili, V., Bowman, D.M.J.S., Bravo, S., Brendel, M.R., Broadley, M.R., Brown, K.A., Bruelheide, H., Brumnich, F., Bruun, H.H., Bruy, D., Buchanan, S.W., Bucher, S.F., Buchmann, N., Buitenwerf, R., Bunker, D.E., Bürger, J., Burrascano, Sabina, Burslem, D.F.R.P., Butterfield, B.J., Byun, C., Marques, M., Scalon, M.C., Caccianiga, M., Cadotte, M., Cailleret, M., Camac, J., Camarero, J.J., Campany, C., Campetella, G., Campos Prieto, Juan Antonio, Cano-Arboleda, L., Canullo, R., Carbognani, M., Carvalho, F., Casanoves, F., Castagneyrol, B., Catford, J.A., Cavender-Bares, J., Cerabolini, Bruno E. L., Cervellini, M., Chacón-Madrigal, E., Chapin, K., Chapin, F.S., Chelli, S., Chen, S.-C., Chen, A., Cherubini, P., Chianucci, F., Choat, B., Chung, K.-S., Chytrý, Milan, Ciccarelli, D., Coll, L., Collins, C.G., Conti, L., Coomes, D., Cornelissen, J.H.C., Cornwell, W.K., Corona, P., Coyea, M., Craine, J., Craven, D., Cromsigt, J.P.G.M., Csecserits, A., Cufar, K., Cuntz, M., and da, Silva, A.C

Abstract

Plant traits the morphological, anatomical, physiological, biochemical and phenological characteristics of plants determine how plants respond to environmental factors, affect other trophic levels, and influence ecosystem properties and their benefits and detriments to people. Plant trait data thus represent the basis for a vast area of research spanning from evolutionary biology, community and functional ecology, to biodiversity conservation, ecosystem and landscape management, restoration, biogeography and earth system modelling. Since its foundation in 2007, the TRY database of plant traits has grown continuously. It now provides unprecedented data coverage under an open access data policy and is the main plant trait database used by the research community worldwide. Increasingly, the TRY database also supports new frontiers of trait-based plant research, including the identification of data gaps and the subsequent mobilization or measurement of new data. To support this development, in this article we evaluate the extent of the trait data compiled in TRY and analyse emerging patterns of data coverage and representativeness. Best species coverage is achieved for categorical traits almost complete coverage for plant growth form . However, most traits relevant for ecology and vegetation modelling are characterized by continuous intraspecific variation and trait environmental relationships. These traits have to be measured on individual plants in their respective environment. Despite unprecedented data coverage, we observe a humbling lack of completeness and representativeness of these continuous traits in many aspects. We, therefore, conclude that reducing data gaps and biases in the TRY database remains a key challenge and requires a coordinated approach to data mobilization and trait measurements. This can only be achieved in collaboration with other initiatives. © 2019 The Authors. Global Change Biology published by John Wiley and Sons Ltd

Published
2020

32. Fetal stem cells obtained from amniotic fluid and wharton's jelly expanded using platelet lysate for tissue engineering applications

Author

Pinto, A. R., Aleixo, I., Cerqueira, M. T., Frias, A. M., Fernandes, S., Rocha, L., Reis, R. L., Neves, N. M., and Universidade do Minho

Subjects
Science & Technology, Tissue engineering, Stem cells
Abstract

Extra-embryonic tissues, such as amniotic fluid (AF) and Wharton´s Jelly (WJ) of umbilical cord, offer many advantages over both embryonic and adult stem cell sources. These tissues are routinely discarded at parturition and the extracorporeal nature of these cell sources facilitates isolation, as well as the comparatively large volume and ease of physical manipulation theoretically increases the number of stem cells that can be isolated. Autologous approaches to use MSCs, namely from bone marrow, have difficulties regarding the limited availability of large amounts of cells from the patient. Fetal stem cells appear to have even more pronounced immunomodulatory properties than adult MSCs (1, 2). This allogeneic escape mechanism may be of therapeutic value, because transplantation of allogeneic human MSCs in stock would be readily available, as opposed to the culture of autologous cells for subsequent transplantation. Cell expansion protocols are based on the use of media supplemented with fetal bovine serum (FBS) as a source of nutrientes and growth factors. The animal serum is not completely safe, once there is a possibility of contamination by animal viroses, prions or others contaminants and it is described that FBS used systematically in MSCs subcultivation induces more humoral immune response (3). Additionally anti-FBS antibodies could be detected in patients after receiving MSCs expanded in FBS (4). Platelet lysate (PL) has enormous possibilities in cell therapy, namely because of the high concentration of growth factors that promotes higher cell expansion, such as tissue regeneration (5). A recent study showed that proliferation of MSCs was much higher on PL gel compared to tissue culture plastic (6). The immunomodulatory properties of MSCs are maintained when expanded in culture medium supplemented with PL (7) Based on these premises we isolated fetal stem cells from AF obtained from amniocentesis and WJ from umbilical cords. These cells were plated and expanded in low density numbers in basal culture medium with FBS or either supplemented with PL. In each passage cells were counted for proliferation kinetics and prepared for flow cytometry analysis. Expanded populations were analysed both population size and complexity and for the MSCs well-known surface markers (CD34, CD45, CD73, CD44, CD106, CD105, CD29, CD90, CD31) and markers related with immune response (HLADR, 80, 83, 86) and embryonic markers SSEA-4 and TRA-1-60.

Published
2012

36. Exercise and Doxorubicin Treatment Modulate Cardiac Mitochondrial Quality Control Signaling.

Author

Marques-Aleixo, I., Santos-Alves, E., Torrella, J. R., Oliveira, P. J., Magalhães, J., and Ascensão, A.

Subjects
EXERCISE physiology, MITOCHONDRIAL pathology, TREADMILL exercise, APOPTOSIS, AUTOPHAGY
Abstract

The cross-tolerance effect of exercise against heart mitochondrial-mediated quality control, remodeling and death-related mechanisms associated with sub-chronic Doxorubicin (DOX) treatment is yet unknown. We therefore analyzed the effects of two distinct chronic exercise models (endurance treadmill training-TM and voluntary free wheel activity-FW) performed during the course of the sub-chronic DOX treatment on mitochondrial susceptibility to permeability transition pore (mPTP), apoptotic and autophagic signaling and mitochondrial dynamics. Male Sprague-Dawley rats were divided into six groups ( n = 6 per group): saline sedentary (SAL + SED), SAL + TM (12-weeks treadmill), SAL + FW (12-weeks voluntary free-wheel), DOX + SED [7-weeks sub-chronic DOX treatment (2 mg kg week)], DOX + TM and DOX + FW. Apoptotic signaling and mPTP regulation were followed by measuring caspase 3, 8 and 9 activities, Bax, Bcl2, CypD, ANT, and cophilin expression. Mitochondrial dynamics (Mfn1, Mfn2, OPA1 and DRP1) and auto(mito)phagy (LC3, Beclin1, Pink1, Parkin and p62)-related proteins were semi-quantified. DOX treatment results in augmented mPTP susceptibility and apoptotic signaling (caspases 3, 8 and 9 and Bax/Bcl2 ratio). Moreover, DOX decreased the expression of fusion-related proteins (Mfn1, Mfn2, OPA1), increased DRP1 and the activation of auto(mito)phagy signaling. TM and FW prevented DOX-increased mPTP susceptibility and apoptotic signaling, alterations in mitochondrial dynamics and inhibits DOX-induced increases in auto(mito)phagy signaling. Collectively, our results suggest that both used chronic exercise models performed before and during the course of sub-chronic DOX treatment limit cardiac mitochondrial-driven apoptotic signaling and regulate alterations in mitochondrial dynamics and auto(mito)phagy in DOX-treated animals. [ABSTRACT FROM AUTHOR]

Published
2018
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39. Apolipoprotein E and undercaboxylated osteocalcin are associated with bone fragility but not with bone mineral density in osteoarthritis patients

Author

Rodrigues⁎, A., primary, Caetano-Lopes, J., additional, Lopes, A., additional, Vale, A.C., additional, Aleixo, I., additional, Perpétuo, I.P., additional, Pena, A.S., additional, Faustino, A., additional, Sepriano, A., additional, Polido-Pereira, J., additional, Vieira-Sousa, E., additional, Vidal, B., additional, Romeu, J.C., additional, Amaral, P.M., additional, Rosa, L.G., additional, da Silva, J.A. Pereira, additional, Monteiro, J., additional, Vaz, M.F., additional, Fonseca, J.E., additional, and Canhão, H., additional

Published
2012
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40. Fracture risk assessed by FRAX translates bone cell activity

Author

Caetano-Lopes, J., primary, Rodrigues, A., additional, Aleixo, I., additional, Perpétuo, I.P., additional, Fernandes, D., additional, Lopes, A., additional, Faustino, A., additional, Pena, A.S., additional, Polido-Pereira, J., additional, Sousa, E., additional, Romeu, J.C., additional, Amaral, P.M., additional, Monteiro, J., additional, Queiroz, M.V., additional, Vaz, M.F., additional, Fonseca, J.E., additional, and Canhão, H., additional

Published
2010
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41. Negative correlation between osteocalcin serum levels and bone mechanical properties

Author

Rodrigues, A., primary, Perpétuo, I.P., additional, Caetano-Lopes, J., additional, Aleixo, I., additional, Pena, A., additional, Faustino, A., additional, Polido-Pereira, J., additional, Sousa, E., additional, Romeu, J., additional, Amaral, P., additional, Monteiro, J., additional, Queiroz, M.V., additional, Vaz, M.F., additional, Fonseca, J.E., additional, and Canhão, H., additional

Published
2010
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42. Absolute risk of fracture calculated by FRAX reflects Bone mechanical properties

Author

Rodrigues, A., primary, Caetano-Lopes, J., additional, Perpétuo, I.P., additional, Aleixo, I., additional, Pena, A.S., additional, Faustino, A., additional, Polido-Pereira, J., additional, Sousa, E., additional, Romeu, J.C., additional, Amaral, P.M., additional, Monteiro, J., additional, Queiroz, M.V., additional, Vaz, M.F., additional, Fonseca, J.E., additional, and Canhão, H., additional

Published
2010
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44. Bone cell activity influences bone fragility and fracture risk

Author

Caetano-Lopes, J, primary, Aleixo, I, additional, Rodrigues, A, additional, Perpetuo, I P, additional, Fernandes, D, additional, Lopes, A, additional, Pena, A, additional, Faustino, A, additional, Vale, M, additional, Amaral, P M, additional, Monteiro, J, additional, Vaz, M F, additional, Canhao, H, additional, and Fonseca, J E, additional

Published
2010
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45. Evaluation of Force Production and Fatigue using an Anaerobic Test Performed by Differently Matured Swimmers.

Author

Soares, S., Silva, R., Aleixo, I., Machado, L., Fernandes, R. J., Maia, J., and Vilas-Boas, J. P.

Subjects
PHYSIOLOGICAL effects of swimming, SWIMMERS, ANAEROBIC capacity, FATIGUE (Physiology), SWIMMING training, SWIMMING techniques
Abstract

The purpose of this research was to characterize fatigue determining thresholds in force-to-time curves produced during 30 second tethered swimming efforts. Ninety swimmers of three maturational groups took part in the study. Values of maximal, mean, and minimum force were determined, as well as the variation coefficient of the mean force. The anaerobic fatigue thresholds were determined on the individual force-totime curves, and typical stroke cycles were determined before and after the fatigue occurrence. Typical force cycles were quantitatively analysed. It was possible to conclude that force tends to increase with maturation in males. In females, tethered force seems to stabilize after puberty. [ABSTRACT FROM AUTHOR]

Published
2010

46. Front Crawl and Backstroke Arm Coordination in Swimmers with Down Syndrome.

Author

Querido, A., Marques-Aleixo, I., Figueiredo, P., Seifert, L., Chollet, D., Vilas-Boas, J.P., Daly, D.J., Corredeira, R., and Fernandes, R.J.

Subjects
SWIMMERS, DOWN syndrome, MOTOR ability, CRAWL stroke (Swimming), BACKSTROKE (Swimming)
Abstract

The aim of this study was to characterize the Index of Arm Coordination (IdC) in swimmers with Down syndrome (DS). The IdC for the front crawl (N=6) was -11.3% ± 5.2% and for the backstroke -13.5 ± 4.8%. This indicates that all swimmers demonstrated a catch-up mode coordination in both strokes. Swimmers with DS did not adapt their arm coordination as usually occurs in swimmers with higher level of proficiency. In front crawl significant positive correlations were found between IdC and the push phase as well as the propulsive phase. An inverse correlation was found between IdC and the non-propulsive phase. In backstroke, the catch-up coordination mode was used by all swimmers. In this study hand lag time values far above those of elite swimmers were observed. There was an inverse relationship between IdC and velocity. [ABSTRACT FROM AUTHOR]

Published
2010

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