Molecular mapping of the regenerative niche in a murine model of myocardial infarction

Adult stem cells are distributed through the whole organism, and present a great potential for the therapy of different types of disease. For the design of efficient therapeutic strategies, it is important to have a more detailed understanding of their basic biological characteristics, as well as of the signals produced by damaged tissues and to which they respond. Myocardial infarction (MI), a disease caused by a lack of blood flow supply in the heart, represents the most common cause of morbidity and mortality in the Western world. Stem cell therapy arises as a promising alternative to conventional treatments, which are often ineffective in preventing loss of cardiomyocytes and fibrosis. Cell therapy protocols must take into account the molecular events that occur in the regenerative niche of MI. In the present study, we investigated the expression profile of ten genes coding for chemokines or cytokines in a murine model of MI, aiming at the characterization of the regenerative niche. MI was induced in adult C57BL/6 mice and heart samples were collected after 24 h and 30 days, as well as from control animals, for quantitative RT-PCR. Expression of the chemokine genes CCL2, CCL3, CCL4, CCL7, CXCL2 and CXCL10 was significantly increased 24 h after infarction, returning to baseline levels on day 30. Expression of the CCL8 gene significantly increased only on day 30, whereas gene expression of CXCL12 and CX3CL1 were not significantly increased in either ischemic period. Finally, expression of the IL-6 gene increased 24 h after infarction and was maintained at a significantly higher level than control samples 30 days later. These results contribute to the better knowledge of the regenerative niche in MI, allowing a more efficient selection or genetic manipulation of cells in therapeutic protocols.

Analysis of 16S rRNA and mxaF genes reveling insights into Methylobacterium niche-specific plant association

The genus Methylobacterium comprises pink-pigmented facultative methylotrophic (PPFM) bacteria, known to be an important plant-associated bacterial group. Species of this group, described as plant-nodulating, have the dual capacity of producing cytokinin and enzymes, such as pectinase and cellulase, involved in systemic resistance induction and nitrogen fixation under specific plant environmental conditions. The aim hereby was to evaluate the phylogenetic distribution of Methylobacterium spp. isolates from different host plants. Thus, a comparative analysis between sequences from structural (16S rRNA) and functional mxaF (which codifies for a subunit of the enzyme methanol dehydrogenase) ubiquitous genes, was undertaken. Notably, some Methylobacterium spp. isolates are generalists through colonizing more than one host plant, whereas others are exclusively found in certain specific plant-species. Congruency between phylogeny and specific host inhabitance was higher in the mxaF gene than in the 16S rRNA, a possible indication of function-based selection in this niche. Therefore, in a first stage, plant colonization by Methylobacterium spp. could represent generalist behavior, possibly related to microbial competition and adaptation to a plant environment. Otherwise, niche-specific colonization is apparently impelled by the host plant.

Il modello green office come nicchia strategica per la transizione sostenibile del sistema universita the green office model as strategic niche for sustainability transition of hei systems

Nella prima parte di questo progetto di tesi, ho analizzato tutte le nozioni teoriche rilevanti in merito alla teoria della transizione. Il primo concetto condiviso in questa trattazione è quello di transizione. Nella parte finale del capitolo, il focus si sposta sul ruolo, in una generica transizione, delle nicchie. Lo strumento centrale in questa struttura sono gli esperimenti di transizione, i quali forniscono un approccio alternativo ai progetti di innovazione classica che sono incentrati nell'ottenimento di soluzioni a breve termine. Vi è dunque una forte relazione tra nicchia e sperimentazione. Infine la trattazione si concentra sul tema dello Strategic Niche Management. Nel secondo capitolo, analizzo il tema della sostenibilità inserita in un contesto universitario. Questa sezione si focalizza sulle strategie di alto livello richieste per dare avvio alla transizione universitaria verso la sostenibilità, identificando gli ostacoli e gli elementi portanti, e definendo una vision al fine di concretizzarla. Il capitolo guida, passo per passo, le università che tentano di mettere in pratica il proprio obiettivo e la vision di sviluppo sostenibile. Una delle problematiche principali per stimare gli sforzi verso la sostenibilità nelle università è costituita in modo particolare dagli strumenti di valutazione. Per questo motivo, è stata sviluppata la valutazione grafica della sostenibilità nell'università (GASU). Al fine di riassumere quanto detto fin qui ed avere un quadro generale più chiaro dell'organizzazione di un campus universitario che mira a diventare sostenibile, ho utilizzato lo strumento gestionale della SWOT Analysis. Negli ultimi due capitoli, infine, analizzo nel dettaglio il modello Green Office. La teorizzazione di questo modello e l'elaborazione dei 6 principi del Green Office sono state effettuate da rootAbility. Le seguenti pagine presentano 3 casi studio di come i 6 principi dei Green Office sono stati adattati alle 3 unità di sostenibilità guidate da studenti e supportate da staff qualificato. L'oggetto della trattazione sono i principali GO affermatisi nei Paesi Bassi. A seguito dell'introduzione del modello relativo al Green Office e dell'illustrazione degli esempi presi in esame, è stato sfruttato lo strumento della feasibility analysis al fine di giudicare se l'idea di business sia praticabile. Il mezzo con cui ho condotto l'analisi sotto riportata è un questionario relativo al modello di Green Office implementato, nel quale viene chiesto di valutare gli aspetti relativi alla organizational feasibility e alla financial feasibility. Infine nella sezione finale ho considerato i Green Office come fossero un unico movimento. L'analisi mira a considerare l'impatto globale del Green Office Movement nei sistemi universitari e come, a seguito del loro consolidarsi nella struttura accademica, possano divenire prassi comune. La struttura proposta contiene elementi sia da il SNM (Strategic Niche Management) che dal TE (Transition Experiment).

Comparative Ultrastructure and Molecular Phylogeny of Selenidium melongena n. sp and S. terebellae Ray 1930 Demonstrate Niche Partitioning in Marine Gregarine Parasites (Apicomplexa)

Gregarine apicomplexans are a diverse group of single-celled parasites that have feeding stages (trophozoites) and gamonts that generally inhabit the extracellular spaces of invertebrate hosts living in marine, freshwater, and terrestrial environments. Inferences about the evolutionary morphology of gregarine apicomplexans are being incrementally refined by molecular phylogenetic data, which suggest that several traits associated with the feeding cells of gregarines arose by convergent evolution. The study reported here supports these inferences by showing how molecular data reveals traits that are phylogenetically misleading within the context of comparative morphology alone. We examined the ultrastructure and molecular phylogenetic positions of two gregarine species isolated from the spaghetti worm Thelepus japonicus: Selenidium terebellae Ray 1930 and S. melongena n. sp. The ultrastructural traits of S. terebellae were very similar to other species of Selenidium sensu stricto, such as having vermiform trophozoites with an apical complex, few epicytic folds, and a dense array of microtubules underlying the trilayered pellicle. By contrast, S. melongena n. sp. lacked a comparably discrete assembly of subpellicular microtubules, instead employing a system of fibrils beneath the cell surface that supported a relatively dense array of helically arranged epicytic folds. Molecular phylogenetic analyses of small subunit rDNA sequences derived from single-cell PCR unexpectedly demonstrated that these two gregarines are close sister species. The ultrastructural differences between these two species were consistent with the fact that S. terebellae infects the inner lining of the host intestines, and S. melongena n. sp. primarily inhabits the coelom, infecting the outside wall of the host intestine. Altogether, these data demonstrate a compelling case of niche partitioning and associated morphological divergence in marine gregarine apicomplexans. (C) 2014 Elsevier GmbH. All rights reserved.

A Stable Niche Assumption-Free Test of Ecological Divergence

Understanding the impact of geological events on diversification processes is central to evolutionary ecology. The recent amalgamation between ecological niche models (ENMs) and phylogenetic analyses has been used to estimate historical ranges of modern lineages by projecting current ecological niches of organisms onto paleoclimatic reconstructions. A critical assumption underlying this approach is that niches are stable over time. Using Notophthalmus viridescens (eastern newt), in which four ecologically diverged subspecies are recognized, we introduce an analytical framework free from the niche stability assumption to examine how refugial retreat and subsequent postglacial expansion have affected intraspecific ecological divergence. We found that the current subspecies designation was not congruent with the phylogenetic lineages. Thus, we examined ecological niche overlap between the refugial and modern populations, in both subspecies and lineage, by creating ENMs independently for modern and estimated last glacial maximum (LGM) newt populations, extracting bioclimate variables by randomly generated points, and conducting principal component analyses. Our analyses consistently showed that when tested as a hypothesis, rather than used as an assumption, the niches of N. viridescens lineages have been unstable since the LGM (both subspecies and lineages). There was greater ecological niche differentiation among the subspecies than the modern phylogenetic lineages, suggesting that the subspecies, rather than the phylogenetic lineages, is the unit of the current ecological divergence. The present study found little evidence that the LGM refugial retreat caused the currently observed ecological divergence and suggests that ecological divergence has occurred during postglacial expansion to the current distribution ranges.

Adaptive behavioural syndromes due to strategic niche specialization

BACKGROUND: Behavioural syndromes, i.e. consistent individual differences in behaviours that are correlated across different functional contexts, are a challenge to evolutionary reasoning because individuals should adapt their behaviour to the requirements of each situation. Behavioural syndromes are often interpreted as a result of constraints resulting in limited plasticity and inflexible behaviour. Alternatively, they may be adaptive if correlated ecological or social challenges functionally integrate apparently independent behaviours. To test the latter hypothesis we repeatedly tested helpers in the cooperative breeder Neolamprologus pulcher for exploration and two types of helping behaviour. In case of adaptive behavioural syndromes we predicted a positive relationship between exploration and aggressive helping (territory defence) and a negative relationship between these behaviours and non-aggressive helping (territory maintenance). RESULTS: As expected, helpers engaging more in territory defence were consistently more explorative and engaged less in territory maintenance, the latter only when dominant breeders were present. Contrary to our prediction, there was no negative relationship between exploration and territory maintenance. CONCLUSION: Our results suggest that the three behaviours we measured are part of behavioural syndromes. These may be adaptive, in that they reflect strategic specialization of helpers into one of two different life history strategies, namely (a) to stay and help in the home territory in order to inherit the breeding position or (b) to disperse early in order to breed independently.

Niche differentiation between diploid and hexaploid Aster amellus

The maintenance of separated diploid and polyploid populations within a contact zone is possible due to both prezygotic and postzygotic isolation mechanisms. Niche differentiation between two cytotypes may be an important prezygotic isolating mechanism and can be studied using reciprocal transplant experiments. We investigated niche differentiation between diploid and hexaploid Aster amellus in their contact zone in the Czech Republic. Diploid populations are confined to habitats with low productivity, whereas hexaploid populations occur in habitats with both low and high productivity. Thus, we chose three diploid populations and six hexaploid populations, three in each of the two different habitat types. We analyzed habitat characteristics and carried out reciprocal transplant experiments in the field using both seeds and adult plants. Sites of diploid and hexaploid populations differed significantly in vegetation and soil properties. The mean number of juveniles was higher at sites of home ploidy level than at sites of foreign ploidy level, suggesting niche differentiation between the two cytotypes. On the other hand, transplanted adult plants survived at all sites and juvenile plants were able to establish at some sites of the foreign cytotype. Furthermore, the mean number of juveniles, survival, and flowering percentages were higher at home sites than at foreign sites, indicating local adaptation. We conclude that niche differentiation between the two cytotypes and local adaptation within each cytotype may contribute to the maintenance of diploid and hexaploid populations of A. amellus in their contact zone. Moreover, further factors, such as differences in flowering phenology and exclusion of minority cytotypes, should also be considered.

Tenascin-C is required for normal Wnt/β-catenin signaling in the whisker follicle stem cell niche.

Whisker follicles have multiple stem cell niches, including epidermal stem cells in the bulge as well as neural crest-derived stem cells and mast cell progenitors in the trabecular region. The neural crest-derived stem cells are a pool of melanocyte precursors. Previously, we found that the extracellular matrix glycoproteins tenascin-C and tenascin-W are expressed near CD34-positive cells in the trabecular stem cell niche of mouse whisker follicles. Here, we analyzed whiskers from tenascin-C knockout mice and found intrafollicular adipocytes and supernumerary mast cells. As Wnt/β-catenin signaling promotes melanogenesis and suppresses the differentiation of adipocytes and mast cells, we analyzed β-catenin subcellular localization in the trabecular niche. We found cytoplasmic and nuclear β-catenin in wild-type mice reflecting active Wnt/β-catenin signaling, whereas β-catenin in tenascin-C knockout mice was mostly cell membrane-associated and thus transcriptionally inactive. Furthermore, cells expressing the Wnt/β-catenin target gene cyclin D1 were enriched in the CD34-positive niches of wild-type compared to tenascin-C knockout mice. We then tested the effects of tenascins on this signaling pathway. We found that tenascin-C and tenascin-W can be co-precipitated with Wnt3a. In vitro, substrate bound tenascins promoted β-catenin-mediated transcription in the presence of Wnt3a, presumably due to the sequestration and concentration of Wnt3a near the cell surface. We conclude that the presence of tenascin-C in whiskers assures active Wnt/β-catenin signaling in the niche thereby maintaining the stem cell pool and suppressing aberrant differentiation, while in the knockout mice with reduced Wnt/β-catenin signaling, stem cells from the trabecular niche can differentiate into ectopic adipocytes and mast cells.