eNOS transfection of adipose-derived stem cells yields bioactive nitric oxide production and improved results in vascular tissue engineering.

This study evaluates the durability of a novel tissue engineered blood vessel (TEBV) created by seeding a natural vascular tissue scaffold (decellularized human saphenous vein allograft) with autologous adipose-derived stem cells (ASC) differentiated into endothelial-like cells. Previous work with this model revealed the graft to be thrombogenic, likely due to inadequate endothelial differentiation as evidenced by minimal production of nitric oxide (NO). To evaluate the importance of NO expression by the seeded cells, we created TEBV using autologous ASC transfected with the endothelial nitric oxide synthase (eNOS) gene to produce NO. We found that transfected ASC produced NO at levels similar to endothelial cell (EC) controls in vitro which was capable of causing vasorelaxation of aortic specimens ex vivo. TEBV (n = 5) created with NO-producing ASC and implanted as interposition grafts within the aorta of rabbits remained patent for two months and demonstrated a non-thrombogenic surface compared to unseeded controls (n = 5). Despite the xenograft nature of the scaffold, the TEBV structure remained well preserved in seeded grafts. In sum, this study demonstrates that upregulation of NO expression within adult stem cells differentiated towards an endothelial-like lineage imparts a non-thrombogenic phenotype and highlights the importance of NO production by cells to be used as endothelial cell substitutes in vascular tissue engineering applications

Role of angiotensin II type 1A receptor phosphorylation, phospholipase D, and extracellular calcium in isoform-specific protein kinase C membrane translocation responses

The angiotensin II type 1A receptor (AT(1A)R) plays an important role in cardiovascular function and as such represents a primary target for therapeutic intervention. The AT(1A)R is coupled via G(q) to the activation of phospholipase C, the hydrolysis of phosphoinositides, release of calcium from intracellular stores, and the activation of protein kinase C (PKC). We show here that PKC beta I and PKC beta II exhibit different membrane translocation patterns in response to AT(1A)R agonist activation. Whereas PKC beta II translocation to the membrane is transient, PKC beta I displays additional translocation responses: persistent membrane localization and oscillations between the membrane and cytosol following agonist removal. The initial translocation of PKC beta I requires the release of calcium from intracellular stores and the activation of phospholipase C, but persistent membrane localization is dependent upon extracellular calcium influx. The mutation of any of the three PKC phosphorylation consensus sites (Ser-331, Ser-338, and Ser-348) localized within the AT(1A)R C-tail significantly increases the probability that persistent increases in diacylglycerol levels and PKC beta I translocation responses will be observed. The persistent increase in AT(1A)R-mediated diacylglycerol formation is mediated by the activation of phospholipase D. Although the persistent PKC beta I membrane translocation response is absolutely dependent upon the PKC activity-dependent recruitment of an extracellular calcium current, it does not require the activation of phospholipase D. Taken together, we show that the patterning of AT(1A)R second messenger response patterns is regulated by heterologous desensitization and PKC isoform substrate specificity

Biodiversity conservation: local and global consequences of the application of “rights of nature” by Ecuador

In 2008, Ecuador recognized the Constitutional Rights of Nature in a global first. This recognition implies a major shift in the human-nature relationship, from one between a subject with agency (humans) and an exploitable object (nature), to a more equilibrated relationship. However, the lack of a standard legal framework has left room for subjective interpretations and variable implementation. The recent widespread concessioning of pristine ecosystems to mining industries has set up an unprecedented conflict and test of these rights. Currently, a landmark case involving Los Cedros Protected Forest and mining companies has reached the Constitutional Court of Ecuador. If Ecuador’s highest Court rules in favor of Los Cedros and the Rights of Nature, it would set a legal precedent with enormous impact on biological conservation. Such a policy shift offers a novel conservation strategy, through citizen oversight and action. A ruling against Los Cedros and the Rights of Nature, while a major setback for biodiversity conservation, would be taken in stride by the active social movement supporting these goals, with the case likely moving into international courts. Meanwhile, extractive activities would continue and expand, with known consequences for biodiversity

Plastid phylogenomics resolves ambiguous relationships within the orchid family and provides a solid timeframe for biogeography and macroevolution

Recent phylogenomic analyses based on the maternally inherited plastid organelle have enlightened evolutionary relationships between the subfamilies of Orchidaceae and most of the tribes. However, uncertainty remains within several subtribes and genera for which phylogenetic relationships have not ever been tested in a phylogenomic context. To address these knowledge-gaps, we here provide the most extensively sampled analysis of the orchid family to date, based on 78 plastid coding genes representing 264 species, 117 genera, 18 tribes and 28 subtribes. Divergence times are also provided as inferred from strict and relaxed molecular clocks and birth–death tree models. Our taxon sampling includes 51 newly sequenced plastid genomes produced by a genome skimming approach. We focus our sampling efforts on previously unplaced clades within tribes Cymbidieae and Epidendreae. Our results confirmed phylogenetic relationships in Orchidaceae as recovered in previous studies, most of which were recovered with maximum support (209 of the 262 tree branches). We provide for the first time a clear phylogenetic placement for Codonorchideae within subfamily Orchidoideae, and Podochilieae and Collabieae within subfamily Epidendroideae. We also identify relationships that have been persistently problematic across multiple studies, regardless of the different details of sampling and genomic datasets used for phylogenetic reconstructions. Our study provides an expanded, robust temporal phylogenomic framework of the Orchidaceae that paves the way for biogeographical and macroevolutionary studies.Universidad de Costa Rica/[814-B8-257]/UCR/Costa RicaUniversidad de Costa Rica/[814-B6-140]/UCR/Costa RicaIDEA WILD/[]//Estados UnidosSociedad Colombiana de Orquideología/[]/SCO/ColombiaFundação de Amparo à Pesquisa do Estado de São Paulo/[11/08308-9]/FAPESP/BrasilFundação de Amparo à Pesquisa do Estado de São Paulo/[13/19124-1]/FAPESP/BrasilSwiss Orchid Foundation/[]//SuizaRoyal Botanic Gardens, Kew/[]//InglaterraSwedish Research Council/[2019-05191]//SueciaSwedish Foundation for Strategic Research/[FFL15-0196]/SSF/SueciaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Jardín Botánico Lankester (JBL

3D Printing: Applications in Evolution and Ecology

In the commercial and medical sectors, 3D printing is delivering on its promise to en‐ able a revolution. However, in the fields of Ecology and Evolution we are only on the brink of embracing the advantages that 3D printing can offer. Here we discuss exam‐ ples where the process has enabled researchers to develop new techniques, work with novel species, and to enhance the impact of outreach activities. Our aim is to showcase the potential that 3D printing offers in terms of improved experimental techniques, greater flexibility, reduced costs and promoting open science, while also discussing its limitations. By taking a general overview of studies using the technique from fields across the broad range of Ecology and Evolution, we show the flexibility of 3D printing technology and aim to inspire the next generation of discoveries

Pollination Biology of the Mushroom-Mimicking Orchid Genus Dracula

Dracula orchids are hypothesized to rely on mushroom mimicry for pollination. These orchids look and smell like mushrooms and are pollinated by mushroom-associated flies in the family Drosophilidae. Dracula includes over 130 species, representing a significant radiation, yet there has never been a systematic study of their pollination biology. Elucidating the processes and mechanisms of pollination in these flowers will broaden our understanding of mimicry within the Orchidaceae, a family well known for its diverse pollination strategies, as well as add to the growing literature on the evolution and maintenance of communication signals. In this study we demonstrate the co-occurrence of the mimics and the putative mushroom models, which is important for evolution by natural selection. We also showed that the resemblance to mushrooms is in fact adaptive, a requisite for floral mimicry. We did this by determining that insect visitors are required for pollination and subsequent fruit set with a hand pollination experiment. We also measured increased visitation rates to the orchids when adjacent to mushrooms. The mechanisms whereby plants attract pollinators can be diverse and often multi-modal, particularly in deceptive systems. Dracula orchids are no exception, with both visual and olfactory signals contributing to the overall success in attracting visitors. We used a series of experiments, first selectively masking the visual and olfactory cues successively, and then using 3D-printed artificial flowers to further disentangle these cues and determine their effect in combination. Upon confirmation that both play a role, we dissected each aspect further. We utilized the artificial flowers to determine the roles of color, contrast, and pattern and employed gas chromatography-mass spectroscopy to identify the volatile signals. The results show that fine-scale contrast is critical to the visual component and that these flowers produce the volatile `mushroom-alcohol' (1-octen-3-ol) in their labella. Finally, we specifically address the hypothesis of brood-site mimicry by using a combination of field observations, insect collections, and rearing studies. The flies gain shelter, a rendezvous location, and food from the flowers. However, no mushroom visiting flies hatched from the flowers, suggesting this may be a brood-site mimicry. This dissertation includes previously unpublished co-authored material.2015-09-2

Causes and process of the decline of Mycenaean culture

Głównym celem pracy, było przedstawienie przyczyn oraz procesu upadku kultury mykeńskiej.Około 1200 p.n.e., większość mykeńskich stanowisk osadniczych, zostało zniszczonych. Ponadto, warstwy zniszczeń datowane na ten okres, zostały odkryte na stanowiskach archeologicznych, w innych rejonach basenu Morza Śródziemnego. Świadczy to o kryzysie na szeroką skalę, którego kulminacja pod koniec epoki brązu, doprowadziła do upadku wielkich cywilizacji tego regionu.Praca składa się z dwóch części. W pierwszej z nich przedstawiono i oceniono, (w oparciu o aktualny stan badań) najpopularniejsze hipotezy tłumaczące upadek kultury mykeńskiej.Druga część pracy skupia się na charakterystyce okresu post-pałacowego, który rozpoczął się po upadku Mykeńskich pałaców, i trwał aż do końca epoki brązu w Regionie Śródziemnomorskim.The main goal of this thesis, is to introduce the causes and process of the decline of Mycenaean culture.Approximately 1200 B.C., most of Mycenaean sites have been destroyed.Moreover, destruction layers with material dated to this period, were founded on other archaeological sites of the Mediterranean Region. It is a proof of wide spread crisis, which culmination in the end of the bronze age, caused the collapse of the greatest civilisations of the region.The thesis consist of two parts. In the first one it has been shown and rated (on the basis of the most relevant data), the most popular hypothesis about the causes of the decline of Mycenaean culture.Second part focuses on the Post-palatial period, which had began after the collapse of the Mycenaean palaces, and have been continuing until the end of the bronze age