Suchergebnisse

The objective of this study was to explore through both in vitro and in vivo experiments the use of reclaimed urban wastewater in dairy cattle production systems with the aim of improving water efficiency and sustainability. Firstly, the use of different tertiary treatments (ultrafiltration (UF), ultraviolet disinfection (UV), chlorination process, and their combination) to improve the quality of an urban secondary effluent was studied in intestinal primary cell cultures evaluating the expression of genes related to apoptosis, cell damage, and inflammation. The results revealed that secondary treated wastewater and waters that were treated with a chlorination process (even tap water) caused an increase in apoptosis, intestinal primary cell damage, and inflammation. The in vivo experiment evaluated the short-term effects on health and performance of using UF- and UV-treated secondary effluent compared with the use of tap water for drinking and preparing milk replacer in young calves from 5 to 47 days of age. Calves previously fed with UF + UV treated secondary effluent clearly preferred tap water when they were exposed to a double water choice at the end of the study. This reduction of the palatability and acceptability was probably due to a greater level of water salinity of the treated reclaimed water (570 vs 1437 ± 76.5 μS/cm of conductivity for tap water and UF-UV treated secondary effluent, respectively), which potentially entailed a reduction of calf concentrate intake (466 vs 351 ± 32.2 g/d for calves fed with tap water and UF-UV treated water, respectively). The use of reclaimed water did not pose an acute risk to animal health. It is concluded that improvements on the tertiary treatment to reduce water salinity should be considered when using reclaimed water for drinking purposes in livestock production systems. This study is a first approach to a more sustainable and efficient use of water in animal husbandry for countries with water scarcity. However, more studies are required before its implementation to further study long-term effects and the presence of new-contaminants not defined in the current legislation. ; info:eu-repo/semantics/acceptedVersion

Microbial communities from harsh environments hold great promise as sources of biotechnologically relevant strains and compounds. In the present work, we have characterized the microorganisms from the supralittoral and splash zone in three different rocky locations of the Western Mediterranean coast, a tough environment characterized by high levels of irradiation and large temperature and salinity fluctuations. We have retrieved a complete view of the ecology and functional aspects of these communities and assessed the biotechnological potential of the cultivable microorganisms. All three locations displayed very similar taxonomic profiles, with the genus Rubrobacter and the families Xenococcaceae, Flammeovirgaceae, Phyllobacteriaceae, Rhodobacteraceae and Trueperaceae being the most abundant taxa; and Ascomycota and halotolerant archaea as members of the eukaryotic and archaeal community respectively. In parallel, the culture‐dependent approach yielded a 100‐isolates collection, out of which 12 displayed high antioxidant activities, as evidenced by two in vitro (hydrogen peroxide and DPPH) and confirmed in vivo with Caenorhabditis elegans assays, in which two isolates, CR22 and CR24, resulted in extended survival rates of the nematodes. This work is the first complete characterization of the Mediterranean splash‐zone coastal microbiome, and our results indicate that this microbial niche is home of an extremophilic community that holds biotechnological potential. ; Financial support from the Spanish Government (Grant Helios, Reference: BIO2015‐66960‐C3‐1‐R co‐financed by FEDER funds and Ministerio de Ciencia, Innovación y Universidades) and from the European CSA on biological standardization BIOROBOOST (EU grant number 820699) is acknowledged. EMM is funded with a Formación de Profesorado Universitario (FPU) grant from the Spanish Government (Ministerio de Ciencia, Innovación y Universidades), with reference FPU17/04184. KT is a recipient of a Doctorado Industrial fellowship from the Ministerio de Ciencia, Innovación y Universidades (Spain), with reference DI‐16‐08976. ÀVV is funded with a Formación de Profesorado Universitario (FPU) grant from the Spanish Government (Ministerio de Ciencia, Innovación y Universidades), with reference FPU18/02578. ; Peer reviewed

Background This article explores the reasons in favour of revising and extending the current 14-day statutory limit to maintaining human embryos in culture. This limit is enshrined in law in over a dozen countries, including the United Kingdom. In two recently published studies (2016), scientists have shown that embryos can be sustained in vitro for about 13 days after fertilisation. Positive reactions to these results have gone hand in hand with calls for revising the 14-day rule, which only allows embryo research until the 14th day after fertilisation. Main text The article explores the most prominent arguments in favour of and against the extension of the 14-day limit for conducting research on human embryos. It situates these arguments within the history of the 14-day limit. I start by discussing the history of the 14-day limit in the United Kingdom and the reasons behind the decision to opt for a compromise between competing moral views. I then analyse the arguments that those who are generally in favour of embryo research put forward in support of extending the 14-day rule, namely (a) the argument of the beneficence of research and (b) the argument of technical feasibility (further explained in the article). I then show how these two arguments played a role in the recent approval of two novel techniques for the replacement of faulty mitochondrial DNA in the United Kingdom. Despite the popularity and widespread use of these arguments, I argue that they are ultimately problematic and should not be straightforwardly accepted (i.e. accepted without further scrutiny). I end by making a case for respecting value pluralism in the context of embryo research, and I present two reasons in favour of respecting value pluralism: the argument of public trust and the argument of democracy. Conclusion I argue that 14-day limit for embryo research is not a valuable tool despite being a solution of compromise, but rather because of it. The importance of respecting value pluralism (and of respecting different views on embryo research) needs to be considered in any evaluation concerning a potential change to the 14-day rule.

International audience ; BACKGROUND:Recently, there has been a re-emergence of cutaneous leishmaniasis in endemic countries and an increase in imported cases in non-endemic countries by travelers, workers, expatriates, immigrants, and military force personnel. Old World cutaneous leishmaniasis is caused primarily by Leishmania major, L. tropica and L. aethiopica. Despite their low sensitivity, diagnosis traditionally includes microscopic and histopathological examinations, and in vitro cultivation. Several conventional PCR techniques have been developed for species identification, which are time-consuming and labour-intensive. Real-time PCR using SYBR green dye, although provides rapid detection, may generate false positive signals. Therefore, a rapid and easy method such as a FRET-based real-time PCR would improve not only the turn-around time of diagnosing Old World cutaneous Leishmania species but will also increase its specificity and sensitivity.METHODS:A FRET-based real-time PCR assay which amplifies the cathepsin L-like cysteine protease B gene encoding a major Leishmania antigen was developed to differentiate L. major, L. tropica, and L. aethiopica in one single step using one set of primers and probes. Assay performance was tested on cutaneous and visceral strains of Leishmania parasite cultures and isolates of other protozoan parasites as well as human biopsy specimen.RESULTS:The assay readily differentiates between the three Old World cutaneous leishmaniasis species based on their melting curve characteristics. A single Tm at 55.2 ± 0.5 °C for L. aethiopica strains was distinguished from a single Tm at 57.4 ± 0.2 °C for L. major strains. A double curve with melting peaks at 66.6 ± 0.1 °C and 48.1 ± 0.5 °C or 55.8 ± 0.6 °C was observed for all L. tropica strains. The assay was further tested on biopsy specimens, which showed 100% agreement with results obtained from isoenzyme electrophoresis and Sanger sequencing.CONCLUSION:Currently, there are no published data on real-time PCR using FRET technology to differentiate between Old World cutaneous Leishmania species. In summary, our assay based on specific hybridization addresses the limitations of previous PCR technology and provides a single step, reliable method of species identification and rapid diagnostic applications.

Facultad de Derecho y Ciencias Sociales. Maestría en Derecho ; On the generality of cultures, families serve as the primary socializing agent between humans. Admittedly, while it has gone through different cycles in the marriage, the nuclear family is interfering remains despite all the changes occurring in society. The role played by the family and marriage in family law, leads to the social and legal support regarding main theme of "non-coital human reproduction", in addition to being maternity fundamental basis in family, plays a leading role in any society, and is essential to legally adjust it according to the scientific, technological advance and the current social reality Artificial procreation, in vitro fertilization or not coital human reproduction, (no doubt difficult to stop and deny at this point phenomenon), acquired the consistency in the Decade of the 60´s, by Australian and English medical equipment. So, as it ushers in a new era of human reproduction unlike the traditional where as everything, comes looking for a solution to a social problem, but that then its effects not were provided both by the legislator as the own research, without leaving aside the acceptance of society since many couples are faced with issues of social and legal, in which not allowed at all have a complete solution to your problem, so we cannot not remain immobile and passive, but we must find more appropriate mechanisms to be able to enter the phenomenon to all fields without having both social and legal implications. No doubt this has obeyed to the problem faced by many couples about infertility and sterility. ; En la generalidad de las culturas, las familias sirven como el agente socializador primario entre los seres humanos. Hay que reconocer que a pesar de que ha pasado por diferentes ciclos en lo que se inmiscuye el matrimonio, la familia nuclear sigue vigente a pesar de todos los cambios que se dan en la sociedad. El papel que juega la familia y el matrimonio dentro del Derecho de familia, conlleva al tema ...

Cell-penetrating protein and its protein transduction domain have been used to deliver drugs and proteins into the cells via receptor-independent endocytosis. A number of cell-penetrating proteins including TAT derived from HIV-1 virus, VP22 from herpes simplex virus and Antennapedia from drosophila have been discovered. Here, we report a cell-penetrating protein, 30Kc19, originating from the hemolymph of silkworm, Bombyx mori. The 30Kc19 is the first cell-penetrating protein that has been found in insect hemolymph. When the 30Kc19 protein produced from recombinant Escherichia coil was supplemented into the medium for mammalian cell culture, 30Kc19 efficiently penetrated into various types of cells and localized at subcellular compartments including mitochondria and cytoplasm. 30Kc19 also delivered cargo proteins such as green fluorescence protein into the cells even though cargo proteins are not able to penetrate into cells by themselves. In addition to the in vitro system, 30Kc19 exhibited the protein transduction property in vivo. When 30Kc19 was intraperitoneally injected into mice, 30Kc19 delivered cargo proteins into various organ tissues of model animals without producing toxicity. Therefore, 30Kc19 has a great potential as a cell-penetrating protein that can be used as a medicinal tool to deliver cargo molecules including proteins into the target organ tissues in the body. (C) 2012 Elsevier Ltd. All rights reserved. ; This work was supported by the National Research Foundation of Korea (NRF) grant funded by the government of Korea (MEST) (No. 2012K0001365, 2012-0000144, 20110020984, 2010-0002520) and by a grant of the Korea Healthcare technology R&D Project, Ministry of Health & Welfare, Republic of Korea. (Grant No. A103017). This work was also supported by the Pioneer Research Program (No. 2012-0001020) and WCU (World Class University) Program (R32-2011-000-10213-0) through the National Research Foundation of Korea (NRF) grant funded by the Ministry of Education, Science, and Technology (MEST). ; OAIID:oai:osos.snu.ac.kr:snu2012-01/102/0000002410/15 ; SEQ:15 ; PERF_CD:SNU2012-01 ; EVAL_ITEM_CD:102 ; USER_ID:0000002410 ; ADJUST_YN:Y ; EMP_ID:A002014 ; DEPT_CD:458 ; CITE_RATE:7.404 ; FILENAME:(2012.12) A protein delivery system using 30Kc19 cell-penetrating protein originating from.pdf ; DEPT_NM:화학생물공학부 ; EMAIL:thpark@snu.ac.kr ; SCOPUS_YN:Y ; CONFIRM:Y

Esta es la versión no revisada del artículo: Valle, J., Burgui, S., Langheinrich, D., Gil, C., Solano, C., Toledo-Arana, A., et al. (2015). Evaluation of Surface Microtopography Engineered by Direct Laser Interference for Bacterial Anti-Biofouling. Macromolecular Bioscience, 15(8), 1060–1069. Se puede consultar la versión publicada en http://doi.org/10.1002/mabi.201500107 ; Biofilm formation by bacterial pathogens on the surface of medical and industrial settings is a 25 serious health problem. Modification of the biomaterial surface topography is a promising 26 strategy to prevent bacterial attachment and biofilm development. However, fabrication of 27 functional biomaterials at large scale with periodic network-topology is still problematic. In this 28 study, we use direct laser interference (DLIP), an easily scalable process, to modify polystyrene 29 surface (PS) topography at sub-micrometer scale. The resulting structure surfaces were 30 interrogated for their capacity to prevent adhesion and biofilm formation of the major human 31 pathogen Staphylococcus aureus. The results revealed that three-dimensional micrometer 32 periodic structures on PS have a profound impact on bacterial adhesion capacity. Thus, line- 33 and pillar-like topographical patterns enhanced S. aureus adhesion, whereas complex lamella 34 microtopography reduced S. aureus adhesion both in static and continuous flow culture 35 conditions. Interestingly, lamella-like textured surfaces retained the capacity to inhibit S. aureus 36 adhesion both when the surface is coated with human serum proteins in vitro and when the 37 material is implanted subcutaneously in a foreign-body associated infection model. Our results 38 establish that the DLIP technology can be used to functionalize polymeric surfaces for the 39 inhibition of bacterial adhesion to surfaces. ; J. Valle was supported by Spanish Ministry of Science and Innovation "Ramón y Cajal" 369 contract. This research was supported by grants AGL2011-23954 and BIO2011-30503-C02-02 370 from the Spanish Ministry of Economy and Competitivity and IIQ14066.RI1 from Innovation 371 Department of the Government of Navarra.

Somatic embryogenesis (SE) provides us a potent biotechnological tool to manipulate the physical and chemical conditions (water availability) along the process and to study their effect in the final success in terms of quantity of somatic embryos produced. In the last years, our research team has been focused on the study of different aspects of the SE in Pinus spp. One of the main aspects affecting SE is the composition of culture media; in this sense, phytohormones play one of the most crucial roles in this propagation system. Many studies in conifers have shown that different stages of SE and somatic embryo development are correlated with distinct endogenous phytohormone profiles under the stress conditions needed for the process (i.e., cytokinins play a regulatory role in stress signaling, which it is essential for radiata pine SE). Based on this knowledge, the aim of this study was to test the effect of different temperatures (18, 23, and 28°C) and gelling agent concentrations (8, 9, and 10 gL-1) during the maturation stage of Pinus radiata SE in maturation and germination rates. Parallel, phytohormone profile of somatic embryos developed was evaluated. In this sense, the highest gellan gum concentration led to significantly lower water availability. At this gellan gum concentration and 23°C a significantly higher number of somatic embryos was obtained and the overall success of the process increased with respect to other treatments assayed. The somatic embryos produced in these conditions showed the highest concentration of iP-type cytokinins and total ribosides. Although, the different conditions applied during maturation of somatic embryos led to different hormonal profiles, they did not affect the ex vitro survival of the resulting somatic plants, where no significant differences were observed. ; This research was funded by MINECO (Spanish Government) project (AGL2016-76143-C4-3R), CYTED (P117RT0522), and DECO (Basque Government). The Ministry of Education, Youth and Sports of the Czechia provided support for this project via the National Program for Sustainability I (LO1204).

Since the European Union (EU) announced their animal testing ban in 2013, all animal experiments related to cosmetics have been prohibited, creating a demand for alternatives to animal experiments for skin studies. Here, we investigated whether an ex vivo live porcine skin model can be employed to study the safety and skin barrier-improving effects of hydroxyacids widely used in cosmetics for keratolytic peels. Glycolic acid (1-10%), salicylic acid (0.2-2%), and lactobionic acid (1.2-12%) were used as representative substances for 慣-hydroxyacid (AHA), 棺-hydroxyacid (BHA), and polyhydroxyacid (PHA), respectively. When hydroxyacids were applied at high concentrations on the porcine skin every other day for 6 days, tissue viability was reduced to 50-80%, suggesting that the toxicity of cosmetic ingredients can be evaluated with this model. Based on tissue viability, the treatment scheme was changed to a single exposure for 20 min. The protective effects of a single exposure of hydroxyacids on skin barrier function were evaluated by examining rhodamine permeability and epidermal structural components of barrier function using immunohistochemistry (IHC) and immunofluorescence (IF) staining. Lactobionic acid (PHAs) improved skin barrier function most compared to other AHAs and BHAs. Most importantly, trans-epidermal water loss (TEWL), an important functional marker of skin barrier function, could be measured with this model, which confirmed the significant skin barrier-protective effects of PHAs. Collectively, we demonstrated that the ex vivo live full-thickness porcine skin model can be an excellent alternative to animal experiments for skin studies on the safety and efficacy of cosmetic ingredients. ; open

The benzothiazepine CGP37157 has shown neuroprotective effects in several in vitro models of excitotoxicity involving dysregulation of intracellular Ca2+ homeostasis. Although its mechanism of neuroprotection is unclear, it is probably related with some of its effects on Ca2+ homeostasis. CGP37157 is a well-known inhibitor of the mitochondrial Na+/Ca2+ exchanger (mNCX). However, it is not very specific and also blocks several other Ca2+ channels and transporters, including voltage-gated Ca2+ channels, plasma membrane Na+/Ca2+ exchanger and the Ca2+ homeostasis modulator 1 channel (CALHM1). In the present work, we have studied if CGP37157 could also induce changes in life expectancy. We now report that CGP37157 extends C. elegans lifespan by 10%–15% with a bell-shaped concentration-response, with high concentrations producing no effect. The effect was even larger (25% increase in life expectancy) in worms fed with heat-inactivated bacteria. The worm CGP37157 concentration producing maximum effect was measured by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and was close to the IC50 for inhibition of the Na+/Ca2+ exchanger. CGP37157 also extended the lifespan in eat-2 mutants (a model for caloric restriction), suggesting that caloric restriction is not involved in the mechanism of lifespan extension. Actually, CGP37157 produced no effect in mutants of the TOR pathway (daf15/unc24) or the insulin/insulin-like growth factor-1 (IGF-1) pathway (daf-2), indicating that the effect involves these pathways. Moreover, CGP37157 was also ineffective in nuo-6 mutants, which have a defect in the mitochondrial respiratory chain complex I. Since it has been described that neuroprotection by this compound in cell cultures is abolished by mitochondrial inhibitors, this suggests that life extension in C. elegans and neuroprotection in cell cultures may share a similar mechanism involving mitochondria. ; This work was supported by grants from the Spanish Ministry of Economy, Industry and Competitiveness, Government of Spain (Ministerio de Economía, Industria y Competitividad, Gobierno de España) to MM and JA (BFU2014-55731-R and BFU2017-83509-R), projects co-financed by the European Union through the European Regional Development Fund, a grant from the Ministry of Education, Board of Castilla y León (Consejería de Educación, Junta de Castilla y León; VA011G18) to JA and a grant from Instituto de Salud Carlos III to CR (Proyectos de Investigación en Salud: PI16/01041, co-financed by FEDER). Some C. elegans strains were provided by the Caenorhabditis Genetics Center (CGC), which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440). JA-V has a fellowship from Junta de Castilla y León (JCyL), co-financed by the Fondo Social Europeo (FSE). PG-C has a FPI fellowship from Ministry of Economy, Industry and Competitiveness. ; Peer reviewed

Testing for embryotoxicity in vitro is an attractive alternative to animal experimentation. The embryonic stem cell test (EST) is such a method, and it has been formally validated by the European Centre for the Validation of Alternative Methods. A number of recent studies have underscored the potential of this method. However, the EST performed well below the 78% accuracy expected from the validation study using a new set of chemicals and pharmaceutical compounds, and also of toxicity criteria, tested to enlarge the database of the validated EST as part of the Work Package III of the ReProTect Project funded within the 6th Framework Programme of the European Union. To assess the performance and applicability domain of the EST we present a detailed review of the substances and their effects in the EST being nitrofen, ochratoxin A, D-penicillamine, methylazoxymethanol, lovastatin, papaverine, warfarin, +¦-aminopropionitrile, dinoseb, furosemide, doxylamine, pravastatin, and metoclopramide. By delineation of the molecular mechanisms of the substances we identify six categories of reasons for misclassifications. Some of these limitations might also affect other in vitro methods assessing embryotoxicity. Substances that fall into these categories need to be included in future validation sets and in validation guidelines for embryotoxicity testing. Most importantly, we suggest conceivable improvements and additions to the EST which will resolve most of the limitations.

This article belongs to the Special Issue Transcriptional and Epigenetic Regulation of Pluripotency and Differentiation. ; Pluripotent stem cells can be stabilized in vitro at different developmental states by the use of specific chemicals and soluble factors. The naïve and primed states are the best characterized pluripotency states. Naïve pluripotent stem cells (PSCs) correspond to the early pre-implantation blastocyst and, in mice, constitute the optimal starting state for subsequent developmental applications. However, the stabilization of human naïve PSCs remains challenging because, after short-term culture, most current methods result in karyotypic abnormalities, aberrant DNA methylation patterns, loss of imprinting and severely compromised developmental potency. We have recently developed a novel method to induce and stabilize naïve human PSCs that consists in the simple addition of a chemical inhibitor for the closely related CDK8 and CDK19 kinases (CDK8/19i). Long-term cultured CDK8/19i-naïve human PSCs preserve their normal karyotype and do not show widespread DNA demethylation. Here, we investigate the long-term stability of allele-specific methylation at imprinted loci and the differentiation potency of CDK8/19i-naïve human PSCs. We report that long-term cultured CDK8/19i-naïve human PSCs retain the imprinting profile of their parental primed cells, and imprints are further retained upon differentiation in the context of teratoma formation. We have also tested the capacity of long-term cultured CDK8/19i-naïve human PSCs to differentiate into primordial germ cell (PGC)-like cells (PGCLCs) and trophoblast stem cells (TSCs), two cell types that are accessible from the naïve state. Interestingly, long-term cultured CDK8/19i-naïve human PSCs differentiated into PGCLCs with a similar efficiency to their primed counterparts. Also, long-term cultured CDK8/19i-naïve human PSCs were able to differentiate into TSCs, a transition that was not possible for primed PSCs. We conclude that inhibition of CDK8/19 stabilizes human PSCs in a functional naïve state that preserves imprinting and potency over long-term culture. ; Work in the laboratory of M.F.F. was funded by the Spanish Association Against Cancer (PROYE18061FERN to M.F.F.), the Asturias Government (PCTI) co-funding 2018-2022/FEDER (IDI/2018/146 to M.F.F.), Fundación General CSIC (0348_CIE_6_E to M.F.F.) and the Health Institute Carlos III (Plan Nacional de I+D+I) co-funding FEDER (PI18/01527 to M.F.F). R.G.U. is supported by the Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER). We also acknowledge support from the IUOPA-ISPA-FINBA (the IUOPA is supported by the Obra Social Cajastur-Liberbank, Spain). Work in the laboratory of M.S. was funded by the IRB and by grants from the Spanish Ministry of Economy co-funded by the European Regional Development Fund (SAF2017-82613-R), ERC (ERC-2014-AdG/669622), Banco Santander (Santander Universities Global Division), la Caixa Foundation, and Secretaria d'Universitats i Recerca del Departament d'Empresa i Coneixement of Catalonia (Grup de Recerca consolidat 2017 SGR 282). ; Peer reviewed

Abstract
There is an ongoing debate on airborne transmission of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) as a risk factor for infection. In this study, the level of SARS-CoV-2 in air and on surfaces of SARS-CoV-2 infected nursing home residents was assessed to gain insight in potential transmission routes. During outbreaks, air samples were collected using three different active and one passive air sampling technique in rooms of infected patients. Oropharyngeal swabs (OPS) of the residents and dry surface swabs were collected. Additionally, longitudinal passive air samples were collected during a period of 4 months in common areas of the wards. Presence of SARS-CoV-2 RNA was determined using RT-qPCR, targeting the RdRp- and E-genes. OPS, samples of two active air samplers and surface swabs with Ct-value ≤35 were tested for the presence of infectious virus by cell culture. In total, 360 air and 319 surface samples from patient rooms and common areas were collected. In rooms of 10 residents with detected SARS-CoV-2 RNA in OPS, SARS-CoV-2 RNA was detected in 93 of 184 collected environmental samples (50.5%) (lowest Ct 29.5), substantially more than in the rooms of residents with negative OPS on the day of environmental sampling (n = 2) (3.6%). SARS-CoV-2 RNA was most frequently present in the larger particle size fractions [>4 μm 60% (6/10); 1–4 μm 50% (5/10); <1 μm 20% (2/10)] (Fischer exact test P = 0.076). The highest proportion of RNA-positive air samples on room level was found with a filtration-based sampler 80% (8/10) and the cyclone-based sampler 70% (7/10), and impingement-based sampler 50% (5/10). SARS-CoV-2 RNA was detected in 10 out of 12 (83%) passive air samples in patient rooms. Both high-touch and low-touch surfaces contained SARS-CoV-2 genome in rooms of residents with positive OPS [high 38% (21/55); low 50% (22/44)]. In one active air sample, infectious virus in vitro was detected. In conclusion, SARS-CoV-2 is frequently detected in air and on surfaces in the immediate surroundings of room-isolated COVID-19 patients, providing evidence of environmental contamination. The environmental contamination of SARS-CoV-2 and infectious aerosols confirm the potential for transmission via air up to several meters.

14 pages, 7 figures, 1 table.-- This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) ; Pulmonary Arterial Hypertension (PAH) is a rare disease caused by the obliteration of the pulmonary arterioles, increasing pulmonary vascular resistance and eventually causing right heart failure. Endothelin-1 (EDN1) is a vasoconstrictor peptide whose levels are indicators of disease progression and its pathway is one of the most common targeted by current treatments. We sequenced the EDN1 untranslated regions of a small subset of patients with PAH, predicted the effect in silico, and used a luciferase assay with the different genotypes to analyze its influence on gene expression. Finally, we used siRNAs against the major transcription factors (TFs) predicted for these regions [peroxisome proliferator-activated receptor γ (PPARγ), Krüppel-Like Factor 4 (KLF4), and vitamin D receptor (VDR)] to assess EDN1 expression in cell culture and validate the binding sites. First, we detected a single nucleotide polymorphism (SNP) in the 5' untranslated region (UTR; rs397751713) and another in the 3'regulatory region (rs2859338) that altered luciferase activity in vitro depending on their genotype. We determined in silico that KLF4/PPARγ could bind to the rs397751713 and VDR to rs2859338. By using siRNAs and luciferase assays, we determined that PPARγ binds differentially to rs397751713. PPARγ and VDR Knock-Down (KD) increased the EDN1 mRNA levels and EDN1 production in porcine aortic endothelial cells (PAECs), while PPARγ and KLF4 KD increased the EDN1 production in HeLa. In conclusion, common variants in EDN1 regulatory regions could alter EDN1 levels. We were able to validate that PPARγ binds in rs397751713 and is a key regulator of EDN1. In addition, KLF4 and VDR regulate EDN1 production in a cell-dependent manner, but VDR does not bind directly to the regions we studied ; This work was funded by the Cardiovascular Research Network of Instituto de Salud Carlos III de Madrid (RD06/0003/0012) Spanish Ministry of Science and Innovation PI18/01233 and Janssen Pharmaceuticals. CINBIO has financial support from Xunta de Galicia and the European Union (European Regional Development Fund-ERDF) (PO FEDER ED431G/02). ML-D and LM-M are supported by a Xunta de Galicia predoctoral fellowship (ED481A-2018/304; IN606A-2020/006). CS is supported by a Ministerio de Universidades FPU predoctoral fellowship (FPU19/00175) ; Peer reviewed

16 pages, 11 figures.-- This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License ; Because of the difficulties in tracking a water mass over time and conducting synoptic surveys over large spatial scales, measurements of biological variables in the ocean are often restricted to isolated Eulerian observations. Here a proof-of-concept of a pseudo-Lagrangian transformation was applied to a time series of chlorophyll-a profiles acquired at a single fixed station to diagnose the shape of a phytoplankton patch, and its physical-biological rates of transformation. During August 2013, a 27-hr time series of observations of horizontal currents, hydrographic properties, turbulent mixing and chlorophyll-a was acquired at a central station of the Ría de Vigo (NW Iberian Peninsula). A chlorophyll-a patch, tilted upward toward the inner part of the Ría, was observed moving back and forth past the sampling station. Its shape and position during the sampling period were modulated by the semi-diurnal (M2) tidal currents and the vertically sheared upwelling circulation. The pseudo-Lagrangian transformation allowed estimation of chlorophyll-a net rates of change. These rates were positive between 12 and 14 m depth, and negative elsewhere, with a mean value of −0.001 ± 0.449 days−1 within the patch maximum. After accounting for the effects of diffusion and sinking, the mean net biological production rate in the upper 15 m (0.53 ± 0.25 days−1) was comparable with in vitro estimates of the balance between phytoplankton growth and mortality obtained from dilution experiments carried out under similar conditions (0.2 ± 0.7 days−1). This pseudo-Lagrangian transformation is complementary to traditional analyses for the quantification of ecological processes and biogeochemical budgets ; This research was funded by projects: STRAMIX (CTM2012-35155) to Miguel Gilcoto and REMEDIOS (CTM2016-75451- C2-1-R) to B. Mouriño-Carballido from the Spanish Ministry of Economy and Competitiveness. M. Villamaña acknowledges the receipt of a FPU fellowship (FPU014/05,385) from the Spanish Ministry of Education, Culture and Sports. B. Fernández Castro was supported by the European Union's Horizon 2020 research and innovation program under the Marie Skłodowska-Curie Grant Agreement No. 834330 (SO-CUP). Funding for open access charge: Universidade de Vigo/CISUG ; Peer reviewed