Cyanovinylation of Aldehydes: Organocatalytic Multicomponent Synthesis of Conjugated Cyanomethyl Vinyl Ethers

A novel organocatalytic multicomponent cyanovinylation of aldehydes was designed for the synthesis of conjugated cyanomethyl vinyl ethers. The reaction was implemented for the synthesis of a 3-substituted 3-(cyanomethoxy)acrylates, using aldehydes as substrates, acetone cyanohydrin as the cyanide anion source, and methyl propiolate as the source of the vinyl component. The multicomponent reaction is catalyzed by N-methyl morpholine (2.5 mol%) to deliver the 3-(cyanomethoxy)acrylates in excellent yields and with preponderance of the E-isomer. The multicomponent reaction manifold is highly tolerant to the structure and composition of the aldehyde (aliphatic, aromatic, heteroaromatics), and it is instrumentally simple (one batch, open atmospheres), economic (2.5 mol% catalyst, stoichiometric reagents), environmentally friendly (no toxic waste), and sustainable (easy scalability).

Delgado-Hernández, Samuel; García-Tellado, Fernando; Tejedor, David

Molecules 26(14), 4120: 1-12 (2021)
DOIDigital.CSIC

Dispersal limitations and long-term persistence drive differentiation from haplotypes to communities within a tropical sky-island: evidence from community metabarcoding

Neutral theory proposes that dispersal stochasticity is one of the main drivers of local diversity. Haplotypes-level genetic variation can now be efficiently sampled from across whole communities, thus making it possible to test neutral predictions from the genetic to species-level diversity, and higher. However, empirical data is still limited, with the few studies to date coming from temperate latitudes. Here, we focus on a tropical mountain within the Transmexican Volcanic Belt to evaluate spatially fine-scale patterns of arthropod community assembly to understand the role of dispersal limitation and landscape features as drivers of diversity. We sampled whole-communities of arthropods for eight orders at a spatial scale ranging from 50 m to 19 km, using whole community metabarcoding. We explored multiple hierarchical levels, from individual haplotypes to lineages at 0.5, 1.5, 3, 5, 7.5% similarity thresholds, to evaluate patterns of richness, turnover, and distance decay of similarity with isolation-by-distance and isolation-by resistance (costs to dispersal given by landscape features) approaches. Our results showed that distance and altitude influence distance decay of similarity at all hierarchical levels. This holds for arthropod groups of contrasting dispersal abilities, but with different strength depending on the spatial scale. Our results support a model where local-scale differentiation mediated by dispersal constraints, combined with long-term persistence of lineages, is an important driver of diversity within tropical sky islands.

Gálvez-Reyes, Nancy; Arribas, Paula; Andújar, Carmelo; Emerson, Brent C.; Piñero, Daniel; Mastretta-Yanes, Alicia

Molecular Ecology (2021)
Digital.CSIC

Metapopulation structure modulates sexual antagonism

Despite the far-reaching evolutionary implications of sexual conflict, the effects of metapopulation structure, when populations are subdivided into several demes connected to some degree by migration, on sexual conflict dynamics are unknown. Here, we used experimental evolution in an insect model system, the seed beetle Callosobruchus maculatus, to assess the independent and interacting effects of selection histories associated with mating system (monogamy vs. polygamy) and population subdivision on sexual conflict evolution. We confirm traditional predictions from sexual conflict theory by revealing increased resistance to male harm in females from populations with a history of intense sexual selection (polygamous populations) compared to females from populations with a history of relaxed sexual selection (monogamous populations). However, selection arising from metapopulation structure reversed the classic pattern of sexually antagonistic coevolution and led to reduced resistance in females from polygamous populations. These results underscore that population spatial structure moderates sexual selection and sexual conflict, and more broadly, that the evolution of sexual conflict is contingent on ecological context. The findings also have implications for population dynamics, conservation biology, and biological control.

Rodríguez-Expósito, Eduardo; García-González, Francisco

Evolution Letters 5(4): 344-358 (2021)
DOIDigital.CSIC

The Bone Regeneration Capacity of BMP-2 + MMP-10 Loaded Scaffolds Depends on the Tissue Status

Biomaterials-mediated bone formation in osteoporosis (OP) is challenging as it requires tissue growth promotion and adequate mineralization. Based on our previous findings, the development of scaffolds combining bone morphogenetic protein 2 (BMP-2) and matrix metalloproteinase 10 (MMP-10) shows promise for OP management. To test our hypothesis, scaffolds containing BMP-2 + MMP-10 at variable ratios or BMP-2 + Alendronate (ALD) were prepared. Systems were characterized and tested in vitro on healthy and OP mesenchymal stem cells and in vivo bone formation was studied on healthy and OP animals. Therapeutic molecules were efficiently encapsulated into PLGA microspheres and embedded into chitosan foams. The use of PLGA (poly(lactic-co-glycolic acid)) microspheres as therapeutic molecule reservoirs allowed them to achieve an in vitro and in vivo controlled release. A beneficial effect on the alkaline phosphatase activity of non-OP cells was observed for both combinations when compared with BMP-2 alone. This effect was not detected on OP cells where all treatments promoted a similar increase in ALP activity compared with control. The in vivo results indicated a positive effect of the BMP-2 + MMP-10 combination at both of the doses tested on tissue repair for OP mice while it had the opposite effect on non-OP animals. This fact can be explained by the scaffold’s slow-release rate and degradation that could be beneficial for delayed bone regeneration conditions but had the reverse effect on healthy animals. Therefore, the development of adequate scaffolds for bone regeneration requires consideration of the tissue catabolic/anabolic balance to obtain biomaterials with degradation/release behaviors suited for the existing tissue status.

García-García, Patricia; Reyes, Ricardo; Rodríguez, José Antonio; Martín, Tomás; Evora, Carmen; Díaz-Rodríguez, Patricia; Delgado, Araceli

Pharmaceutics 13(7), 979: 1-23 (2021)
DOIDigital.CSIC

Changes in the structure of seed dispersal networks when including interaction outcomes from both plant and animal perspectives

Interaction frequency is the most common currency in quantitative ecological networks, although interaction quality can also affect benefits provided by mutualisms. Here, we evaluate if interaction quality can modify network topology, species' role and whether such changes affect community vulnerability to species loss. We use a well-examined study system (bird–lizard and fleshy-fruited plants in the ‘thermophilous' woodland of the Canary Islands) to compare network and species-level metrics from a network based on fruit consumption rates (interaction frequency, IF), against networks reflecting functional outcomes: a seed dispersal effectiveness network (SDE) quantifying recruitment, and a fruit resource provisioning network (FRP), accounting for the nutrient supply of fruits. Nestedness decreased in the FRP and the SDE networks, due to the lack of association between fruit consumption rates and 1) nutrient content and; 2) recruitment at the seed deposition sites, respectively. The FRP network showed lower niche overlap due to resource use complementarity among frugivores. Interaction evenness was lower in the SDE network, in response to a higher dominance of lizards in the recruitment of heliophilous species. Such changes, however, did not result in enhanced vulnerability against extinctions. At the plant species level, strength changed in the FRP network in frequently consumed or highly nutritious species. The number of effective partners decreased for species whose seeds were deposited in unsuitable places for recruitment. In frugivores, strength was consistent across networks (SDE vs IF), showing that consumption rates outweighed differences in dispersal quality. In the case of lizards, the increased importance of nutrient-rich species resulted in a higher number of effective partners.

Our work shows that although frequency strongly impacts interaction effects, accounting for quality improves our inferences about interaction assembly and species role. Thus, future studies including interaction outcomes from both partners' perspectives will provide valuable insights about the net effects of mutualistic interactions.

González-Castro, Aaron; Morán-López, Teresa; Nogales, Manuel; Traveset, Anna

Oikos (2021)
DOIDigital.CSIC

Acrylonitrile Derivatives against Trypanosoma cruzi: In Vitro Activity and Programmed Cell Death Study

The neglected infection known as Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, results in more than 7000 deaths per year, with an increasing number of cases in non-endemic areas such as Europe or the United States. Moreover, with the current available therapy, only two compounds which are active against the acute phase of the disease are readily available. In addition, these therapeutic agents display multiple undesired side effects such as high toxicity, they are expensive, the treatment is lengthy and the resistant strain has emerged. Therefore, there is a need to find new compounds against Chagas disease which should be active against the parasite but also cause low toxicity to the patients. In the present work, the activity of novel acrylonitriles against Trypanosoma cruzi was evaluated as well as the analysis of the physiological events induced in the treated parasites related to the cell death process. Hence, the characteristic features of an apoptosis-like process such as chromatin condensation and mitochondrial membrane potential, among others, were studied. From the 32 compounds tested against the epimastigote stage of T. cruzi, 11 were selected based on their selectivity index to determine if these compounds were able to induce programmed cell death (PCD) in the treated parasites. Furthermore, acrylonitriles Q5, Q7, Q19, Q27 and Q29 were shown to trigger physiological events related in the PCD. Therefore, this study highlights the therapeutic potential of acrylonitriles as novel trypanocidal agents.

Bethencourt-Estrella, Carlos J.; Delgado-Hernández, Samuel; López-Arencibia, Atteneri; San Nicolás-Hernández, Desirée; Sifaoui, Ines; Tejedor, David; García-Tellado, Fernando; Lorenzo-Morales, Jacob; Piñero, José E.

Pharmaceuticals 14(6), 552: 1-19 (2021)
DOIDigital.CSIC

Dynamic Nucleophilic Aromatic Substitution of Tetrazines

A dynamic nucleophilic aromatic substitution of tetrazines (SNTz) is presented herein. It combines all the advantages of dynamic covalent chemistry with the versatility of the tetrazine moiety. Indeed, libraries of compounds or sophisticated molecular structures can be easily obtained, which are susceptible to post-functionalization by inverse electron demand Diels–Alder (IEDDA) reaction, which also locks the exchange. Additionally, the structures obtained can be disassembled upon the application of the right stimulus, either UV irradiation or a suitable chemical reagent. Moreover, SNTz is compatible with the imine chemistry of anilines. The high potential of this methodology has been proved by building two responsive supramolecular systems: A macrocycle that displays a light-induced release of acetylcholine; and a truncated [4+6] tetrahedral shape-persistent fluorescent cage, which is disassembled by thiols unless it is post-stabilized by IEDDA.

Santos, Tanausú; Rivero, David S.; Pérez-Pérez, Yaiza; Martín-Encinas, Endika; Pasán, Jorge; Hernández Daranas, Antonio; Carillo, Romen

Angewandte Chemie - International Edition 60(34): 18783-18791 (2021)
DOIDigital.CSIC

The Chemistry of Reactive Oxygen Species (ROS) Revisited: Outlining Their Role in Biological Macromolecules (DNA, Lipids and Proteins) and Induced Pathologies

Living species are continuously subjected to all extrinsic forms of reactive oxidants and others that are produced endogenously. There is extensive literature on the generation and effects of reactive oxygen species (ROS) in biological processes, both in terms of alteration and their role in cellular signaling and regulatory pathways. Cells produce ROS as a controlled physiological process, but increasing ROS becomes pathological and leads to oxidative stress and disease. The induction of oxidative stress is an imbalance between the production of radical species and the antioxidant defense systems, which can cause damage to cellular biomolecules, including lipids, proteins and DNA. Cellular and biochemical experiments have been complemented in various ways to explain the biological chemistry of ROS oxidants. However, it is often unclear how this translates into chemical reactions involving redox changes. This review addresses this question and includes a robust mechanistic explanation of the chemical reactions of ROS and oxidative stress.

Andrés Juan, Celia; Pérez de Lastra, José Manuel; Plou Gasca, Francisco José; Pérez-Lebeña, Euardo

International Journal of Molecular Sciences 22(9), 4642: 1-21(2021)
DOIDigital.CSIC

Biodiversity monitoring using environmental DNA

Monitoring biodiversity is essential to protect, preserve and restore ecosystems, particularly in the context of current challenges such as climate change, habitat destruction and globalization (Baird & Hajibabaei, 2012). Biomonitoring is needed for developing biotic indices for assessing ecological status, measuring impacts of anthropogenic activities in natural ecosystems, evaluating biodiversity loss, surveying nonindigenous species, conservation, and identifying cryptic species (Balvanera et al., 2006; Fišer et al., 2018). Thus, spatially and temporally structured biomonitoring activities provide a powerful tool for the implementation of regional, national and international regulations, directives and policies for nature conservation. However substantial impediments exist including access to remote locations, limited specialist taxonomic knowledge, cost, slow pace of human-driven data analyses, and typically low sensitivity for detection of rare and elusive species (Zinger et al., 2020). These drawbacks are often translated into expensive monitoring activities with limited spatial, temporal and taxonomic coverage. In this context, new approaches for biomonitoring are being explored, among which advanced DNA-based technologies are emerging (Kissling et al., 2018). The field of biodiversity monitoring through the analysis of the pool of DNA isolated from environmental samples, referred to as environmental DNA or eDNA (Pawlowski et al., 2020; Taberlet et al., 2012), is rapidly growing. This growth is being driven through improved approaches for sampling, data generation and analyses, and with recent advances on how eDNA should be interpreted for biodiversity assessments (Bohmann et al., 2014). The success of eDNA-based biomonitoring is reflected in exponential growth of publications within this area and increasing submissions to Molecular Ecology Resources in particular (Figure 1). Molecular Ecology Resources aims to publish high quality eDNA studies that serve as broad resources, including innovative methodologies for DNA sampling, enhanced laboratory protocols for data generation, or new computer programs and statistical advances for data analyses. Thus, the aim of this editorial is to contribute to producing good quality DNA data-derived essential biodiversity variables (EBVs) (Kissling et al., 2018) by providing guidance to the community submitting articles on the subject. For that purpose, we have summarized best practices established in published literature related to the different phases involved in the process, namely sampling, laboratory work, bioinformatic analyses and data interpretation (Figure 2).

Rodríguez-Ezpeleta, Naiara; Zinger, Lucie; Kinziger, Andrew; Bik, Holly M.; Bonin, Aurélie; Coissac, Eric; Emerson, Brent C. ; Martins Lopes, Carla; Pelletier, Tara A.; Taberlet, Pierre; Narum, Shawn

Molecular Ecology Resources 21(5): 1405-1409 (2021)
DOIDigital.CSIC

FLTX2: A Novel Tamoxifen Derivative Endowed with Antiestrogenic, Fluorescent, and Photosensitizer Properties

Tamoxifen is the most widely used selective modulator of estrogen receptors (SERM) and the first strategy as coadjuvant therapy for the treatment of estrogen-receptor (ER) positive breast cancer worldwide. In spite of such success, tamoxifen is not devoid of undesirable effects, the most life-threatening reported so far affecting uterine tissues. Indeed, tamoxifen treatment is discouraged in women under risk of uterine cancers. Recent molecular design efforts have endeavoured the development of tamoxifen derivatives with antiestrogen properties but lacking agonistic uterine tropism. One of this is FLTX2, formed by the covalent binding of tamoxifen as ER binding core, 7-nitrobenzofurazan (NBD) as the florescent dye, and Rose Bengal (RB) as source for reactive oxygen species. Our analyses demonstrate (1) FLTX2 is endowed with similar antiestrogen potency as tamoxifen and its predecessor FLTX1, (2) shows a strong absorption in the blue spectral range, associated to the NBD moiety, which efficiently transfers the excitation energy to RB through intramolecular FRET mechanism, (3) generates superoxide anions in a concentration- and irradiation time-dependent process, and (4) Induces concentration- and time-dependent MCF7 apoptotic cell death. These properties make FLTX2 a very promising candidate to lead a novel generation of SERMs with the endogenous capacity to promote breast tumour cell death in situ by photosensitization.

Díaz, Mario; Lobo, Fernando; Hernández, Dácil; Amesty, Ángel; Valdés-Baizabal, Catalina; Canerina-Amaro, Ana; Mesa-Herrera, Fátima; Soler, Kevin; Boto, Alicia; Marín, Raquel; Estévez-Braun, Ana; Lahoz, Fernando

International Journal of Molecular Sciences 22(10), 5339: 1-21 (2021)
DOIDigital.CSIC