Electroactive poly(vinylidene fluoride-trifluoroethylene)/graphene composites for cardiac tissue engineering applications.

Electroactive materials are increasingly being used in strategies to regenerate cardiac tissue. These materials, particularly those with electrical conductivity, are used to actively recreate the electromechanical nature of the cardiac tissue. In the present work, we describe a novel combination of poly(vinylidene fluoride-trifluoroethylene) (P(VDF-TrFE)), a highly electroactive polymer, with graphene (G), exhibiting high electrical conductivity. G/P(VDF-TrFE) films have been characterized in terms of topographical, physico-chemical, mechanical, electrical, and thermal properties, and studied the response of cardiomyocytes adhering to them. The results indicate that the crystallinity and the wettability of the composites remain almost unaffected after G incorporation. In turn, surface roughness, Young modulus, and electric properties are higher in G/P(VDF-TrFE). Finally, the composites are highly biocompatible and able to support cardiomyocyte adhesion and proliferation, particularly surface treated ones, demonstrating the suitability of these materials for cardiac tissue engineering applications.

Positive species interactions strengthen in a high-CO2 ocean.

Negative interactions among species are a major force shaping natural communities and are predicted to strengthen as climate change intensifies. Similarly, positive interactions are anticipated to intensify and could buffer the consequences of climate-driven disturbances. We used in situ experiments at volcanic CO2 vents within a temperate rocky reef to show that ocean acidification can drive community reorganization through indirect and direct positive pathways. A keystone species, the algal-farming damselfish Parma alboscapularis, enhanced primary productivity through its weeding of algae whose productivity was also boosted by elevated CO2. The accelerated primary productivity was associated with increased densities of primary consumers (herbivorous invertebrates), which indirectly supported increased secondary consumers densities (predatory fish) (i.e. strengthening of bottom-up fuelling). However, this keystone species also reduced predatory fish densities through behavioural interference, releasing invertebrate prey from predation pressure and enabling a further boost in prey densities (i.e. weakening of top-down control). We uncover a novel mechanism where a keystone herbivore mediates bottom-up and top-down processes simultaneously to boost populations of a coexisting herbivore, resulting in altered food web interactions and predator populations under future ocean acidification.

Trophic pyramids reorganize when food web architecture fails to adjust to ocean change.

As human activities intensify, the structures of ecosystems and their food webs often reorganize. Through the study of mesocosms harboring a diverse benthic coastal community, we reveal that food web architecture can be inflexible under ocean warming and acidification and unable to compensate for the decline or proliferation of taxa. Key stabilizing processes, including functional redundancy, trophic compensation, and species substitution, were largely absent under future climate conditions. A trophic pyramid emerged in which biomass expanded at the base and top but contracted in the center. This structure may characterize a transitionary state before collapse into shortened, bottom-heavy food webs that characterize ecosystems subject to persistent abiotic stress. We show that where food web architecture lacks adjustability, the adaptive capacity of ecosystems to global change is weak and ecosystem degradation likely.

Functional loss in herbivores drives runaway expansion of weedy algae in a near-future ocean.

The ability of a community to absorb environmental change without undergoing structural modification is a hallmark of ecological resistance. The recognition that species interactions can stabilize community processes has led to the idea that the effects of climate change may be less than what most considerations currently allow. We tested whether herbivory can compensate for the expansion of weedy algae triggered by CO2 enrichment and warming. Using a six-month mesocosm experiment, we show that increasing per capita herbivory by gastropods absorbs the boosted effects of CO2 enrichment on algal production in temperate systems of weak to moderate herbivory. However, under the combined effects of acidification and warming this compensatory effect was eroded by reducing the diversity, density and biomass of herbivores. This loss of functionality combined with boosted primary productivity drove a fourfold expansion of weedy algal species. Our results demonstrate capacity to buffer ecosystems against CO2 enrichment, but loss of this capacity through ocean warming either in isolation or combined with CO2, driving significant algal turf expansion. Identifying compensatory processes and the circumstances under which they prevail could potentially help manage the impacts of ocean warming and acidification, which are further amplified by local disturbances such as habitat loss and herbivore over-exploitation.

A triple trophic boost: How carbon emissions indirectly change a marine food chain.

The pervasive enrichment of CO2 in our oceans is a well-documented stressor to marine life. Yet, there is little understanding about how CO2 affects species indirectly in naturally complex communities. Using natural CO2 vents, we investigated the indirect effects of CO2 enrichment through a marine food chain. We show how CO2 boosted the biomass of three trophic levels: from the primary producers (algae), through to their grazers (gastropods), and finally through to their predators (fish). We also found that consumption by both grazers and predators intensified under CO2 enrichment, but, ultimately, this top-down control failed to compensate for the boosted biomass of both primary producers and herbivores (bottom-up control). Our study suggests that indirect effects can buffer the ubiquitous and direct, negative effects of CO2 enrichment by allowing the upward propagation of resources through the food chain. Maintaining the natural complexity of food webs in our ocean communities could, therefore, help minimize the future impacts of CO2 enrichment.

CO2 emissions boost the benefits of crop production by farming damselfish.

Farming is a technique employed by both humans and animals to enhance crop yields, allowing their populations to increase beyond the natural carrying capacity of the environment. Using volcanic CO2 vents, we investigate how a species of herbivorous fish (the black scalyfin Parma alboscapularis) may use increasing anthropogenic CO2 emissions to enhance its crop yields. We found that these farming fish can take advantage of this resource enrichment, to grow crops within smaller territories and increase the capacity of the environment to support more densely packed fish populations.

Microhabitat change alters abundances of competing species and decreases species richness under ocean acidification.

Niche segregation allows competing species to capture resources in contrasting ways so they can co-exist and maintain diversity, yet global change is simplifying ecosystems and associated niche diversity. Whether climate perturbations alter niche occupancy among co-occurring species and affect species diversity is a key, but unanswered question. Using CO2 vents as natural analogues of ocean acidification, we show that competing fish species with overlapping diets are partially segregated across microhabitat niches and differently-orientated substrata under ambient CO2 conditions. Under elevated CO2, benthic microhabitats experienced a significant increase in non-calcifying turf and fleshy algae but a sharp reduction in calcareous algae. The increased availability of turf and fleshy algae supported increased densities of a competitively dominant species, whilst the reduction in calcifying algal microhabitats decreased densities of several subordinate species. The change in microhabitat availability also drove an increased overlap in microhabitat use among competing fishes at the vents, associated with a reduced fish species richness on horizontal substrates. We conclude that loss of preferred microhabitat niches, exacerbated by population proliferation of competitively dominant species, can drive population losses of less common and subordinate species, and reduce local species richness. The indirect effects of ocean acidification on microhabitat availability can therefore impair maintenance of species populations, and drive changes in local community and biodiversity patterns.

Climate change could drive marine food web collapse through altered trophic flows and cyanobacterial proliferation.

Global warming and ocean acidification are forecast to exert significant impacts on marine ecosystems worldwide. However, most of these projections are based on ecological proxies or experiments on single species or simplified food webs. How energy fluxes are likely to change in marine food webs in response to future climates remains unclear, hampering forecasts of ecosystem functioning. Using a sophisticated mesocosm experiment, we model energy flows through a species-rich multilevel food web, with live habitats, natural abiotic variability, and the potential for intra- and intergenerational adaptation. We show experimentally that the combined stress of acidification and warming reduced energy flows from the first trophic level (primary producers and detritus) to the second (herbivores), and from the second to the third trophic level (carnivores). Warming in isolation also reduced the energy flow from herbivores to carnivores, the efficiency of energy transfer from primary producers and detritus to herbivores and detritivores, and the living biomass of detritivores, herbivores, and carnivores. Whilst warming and acidification jointly boosted primary producer biomass through an expansion of cyanobacteria, this biomass was converted to detritus rather than to biomass at higher trophic levels-i.e., production was constrained to the base of the food web. In contrast, ocean acidification affected the food web positively by enhancing trophic flow from detritus and primary producers to herbivores, and by increasing the biomass of carnivores. Our results show how future climate change can potentially weaken marine food webs through reduced energy flow to higher trophic levels and a shift towards a more detritus-based system, leading to food web simplification and altered producer-consumer dynamics, both of which have important implications for the structuring of benthic communities.

Species Interactions Drive Fish Biodiversity Loss in a High-CO2 World.

Accelerating climate change is eroding the functioning and stability of ecosystems by weakening the interactions among species that stabilize biological communities against change [1]. A key challenge to forecasting the future of ecosystems centers on how to extrapolate results from short-term, single-species studies to community-level responses that are mediated by key mechanisms such as competition, resource availability (bottom-up control), and predation (top-down control) [2]. We used CO2 vents as potential analogs of ocean acidification combined with in situ experiments to test current predictions of fish biodiversity loss and community change due to elevated CO2 [3] and to elucidate the potential mechanisms that drive such change. We show that high risk-taking behavior and competitive strength, combined with resource enrichment and collapse of predator populations, fostered already common species, enabling them to double their populations under acidified conditions. However, the release of these competitive dominants from predator control led to suppression of less common and subordinate competitors that did not benefit from resource enrichment and reduced predation. As a result, local biodiversity was lost and novel fish community compositions were created under elevated CO2. Our study identifies the species interactions most affected by ocean acidification, revealing potential sources of natural selection. We also reveal how diminished predator abundances can have cascading effects on local species diversity, mediated by complex species interactions. Reduced overfishing of predators could therefore act as a key action to stall diversity loss and ecosystem change in a high-CO2 world. VIDEO ABSTRACT.

Boosted food web productivity through ocean acidification collapses under warming.

Future climate is forecast to drive bottom-up (resource driven) and top-down (consumer driven) change to food web dynamics and community structure. Yet, our predictive understanding of these changes is hampered by an over-reliance on simplified laboratory systems centred on single trophic levels. Using a large mesocosm experiment, we reveal how future ocean acidification and warming modify trophic linkages across a three-level food web: that is, primary (algae), secondary (herbivorous invertebrates) and tertiary (predatory fish) producers. Both elevated CO2 and elevated temperature boosted primary production. Under elevated CO2 , the enhanced bottom-up forcing propagated through all trophic levels. Elevated temperature, however, negated the benefits of elevated CO2 by stalling secondary production. This imbalance caused secondary producer populations to decline as elevated temperature drove predators to consume their prey more rapidly in the face of higher metabolic demand. Our findings demonstrate how anthropogenic CO2 can function as a resource that boosts productivity throughout food webs, and how warming can reverse this effect by acting as a stressor to trophic interactions. Understanding the shifting balance between the propagation of resource enrichment and its consumption across trophic levels provides a predictive understanding of future dynamics of stability and collapse in food webs and fisheries production.

Testis peritubular myoid cells increase their motility and express matrix-metalloproteinase 9 (MMP-9) after interaction with embryonal carcinoma cells.

Today cancer research studies have highlighted the role of the cancer-stroma interaction in the regulation of invasive processes. However, very little is known about cell-to-cell relationships between germinal cancer cells and the somatic ones belong to their close environment, particularly at early invasion stages. Here, we have studied the potential role of the seminiferous peritubular myoid cells (PTCs), as potential part of the reactive stroma, like tumor myofibroblast, in the progression of embryonal carcinoma (EC). To this end, we show results on the in vitro interactions between F9 murine embryonal carcinoma cells (EC cells) and primary cultures of murine PTCs, using contact-dependent and contact-independent 2D co-cultures. In these circumstances, when EC cells interact with PTCs they change their migratory behavior and matrix-metalloproteinase 9 (MMP-9) was up-regulated in PTCs. Additionally, among a variety of cytokines implicated in tumor-stroma cross-talk, we have examined in more detail the influence of tumor necrosis factor alpha (TNF-α). In this regard, it was observed that this cytokine induced a MMP-9 secretion by PTCs in a pattern dependent on its concentration, whereas does not increase the migration capacity of cancer cells. All together, our results provide evidence for a role played by peritubular myoid cells and cancer-cell secreted TNF- α for a change in the tumor microenvironment during the early stages of EC progression.

Evidence for a role of matrix metalloproteinases and their inhibitors in primordial germ cell migration.

Understanding the mechanisms that enable migrating cells to reach their targets is of vital importance, as several pathologies, including cardiac defects and some tumours, are consequences of altered cell migration. With a view to evaluating if matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play a role in the active migration of primordial germ cells (PGCs) from their place of origin in extra-embryonic sites towards their final destination in the developing gonads, we analysed the expression of mRNAs encoding nine MMPs and four TIMPs in migrating (E10.5) and post-migrating (E12.5) PGCs by means of quantitative polymerase chain reaction and the presence of MT1-MMP in the membrane of these cells. Our results show that PGCs express MMP-2, MMP-9, MMP-11, MT1-MMP, TIMP-1, TIMP-2 and TIMP-3 at both migrating and non-migrating stages. Comparing expression levels of MMP genes between E10.5 and E12.5 PGCs revealed higher expression in migrating PGCs of MT1- MMP (10.3-fold), MMP-2 (4.8-fold), MMP-11 (3.2-fold) and MMP-9 (2.1-fold). Similarly, the levels of TIMP gene expression were always higher in E12.5 genital ridge somatic cells: TIMP-3 (3.4-fold), TIMP-1 (2.4-fold) and TIMP-2 (1.8-fold). Moreover, the analysis at protein level showed the presence of MT1-MMP in the membrane of migrating PGCs whereas the expression of these metalloproteinase is not detected once the PGCs have reach the urogenital ridges and stop migrating. These results suggest that the change from the motile to non-motile phenotype that occurs during PGC maturation to gonocytes may be mediated in part by enhanced expression of MMPs in migrating PGCs together with higher expression of TIMPs in E12.5 genital ridges.

Peritubular myoid cell-derived factors and its potential role in the progression of testicular germ cell tumours.

The tumour surrounding stroma, known as reactive stroma, is a crucial factor to understand cancer cell growth and invasion. In the normal adult testis, the stroma contains extracellular matrix components, fibroblasts, infiltrating leucocytes, lymphocytes, macrophages and capillaries, as well as other specific cell populations, like Leydig cells and a thin myoepithelium surrounding the seminiferous tubules constituted by the peritubular cells. All these cells are an important source of proliferation and survival promoting signals, proteolytic enzymes, migratory cues and pro-angiogenic factors. Ascribable to this pro-invasive activity, the tumour reactive stroma cells, especially cancer-associated myofibroblasts, have emerged as a promising target for cancer therapy. This review is focused on the potential role of the peritubular myoid cells in the development of testicular germ cell tumours as the precursors of cancer-associated myofibroblast and on an experimental model for the study of testis germinal cancer stroma and on the differences between normal and tumour-associated stromal cells, including the molecular mechanisms that mediate the important cancer stroma crosstalk. Special attention will be paid to the cancer-associated myofibroblasts as possible therapeutic targets, because they are one of the main components of the reactive stroma and are known to secrete a variety of paracrine factors that stimulate tumour progression.

Vascularization of testicular germ cell tumours: evidence from experimental teratocarcinomas.

Testicular germ cell tumours (TGCTs) represent about 2% of male malignancies, being the most common cancer among adolescents and young adults. As in most neoplasias, TGCTs show a chaotic vascular architecture, altered blood supply and over-expression of pro-angiogenic factors, aspects closely related to tumour overgrowth and metastasis. Following this trend, our laboratory has analysed the effect of the hypoxic tumour microenvironment on cancer stem cells, particularly the expression of factors related to vascularization, such as matrix metalloproteinases, adhesion molecules, vascular endothelial growth factors (VEGF) and VEGF receptors. This review also summarizes our present knowledge on vascularization in the normal and neoplastic testis, the potential role of the factors involved in TGCT neovascularization and their importance as possible therapeutic targets.

Angiogenesis and vascular network of teratocarcinoma from embryonic stem cell transplant into seminiferous tubules.

BACKGROUND: Carcinoma in situ (CIS) of the testis is considered to be a precancerous germinal cell lesion, but the precise cellular and molecular mechanisms underlying transformation of CIS into invasive pluripotent cancer cells remain to be elucidated. Moreover, a satisfactory animal model for the experimental study of germinal tumours has not been developed to date. METHODS: We have developed a tumour model that involves the microinjection of green fluorescent protein-labelled embryonic stem (ES) cells (which are functionally equivalent to CIS cells) into syngenic mouse seminiferous tubules, a unique cell microenvironment in which germinal cells mature and CIS arise. To characterise the vascularisation of teratocarcinomas, which arise after cell transplant, we used immunohistochemistry, together with a qualitative and quantitative analysis of scanning electron microscopy images of corrosion casting samples. RESULTS: Embryonic stem cells transplanted into seminiferous tubules did not differentiate into germinal cells, but rather they behaved as invasive embryonal carcinoma (EC) stem cells. The vascular pattern of the experimental teratocarcinomas showed a highly disorganised architecture, and some of the neoplastic capillaries were derived, at least in part, from the original transplanted ES cells. CONCLUSION: The transplantation of pluripotent ES cells into seminiferous tubules efficiently recapitulates the early stages of development of teratocarcinomas. Consequently, this method constitutes a novel in vivo model to study the mechanisms of invasion and progression of experimental germinal tumours.