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Assessing the Impact of Pumpkins Plantation, Harvest and Storage Decisions on a Collaborative Supply Chain with Data Analysis Tools

[EN] Successful pumpkins production requires the use of varieties that jointly with other factors yield well and produce pumpkins of the size, shape, color, and quality demanded by the market. But not only these issues are important. The perishable nature of pumpkins makes other issues such as how to prevent deterioration after harvest to become also relevant. In this paper the pumpkins plantation, harvest and storage (PHS) process is described and how some decisions affect certain goals, such as yield or conservation time. Additionally, some decision-making insights in a supply chain collaborative scenario made up of two stages: plantation/harvest and storage are given, where yield and conservation time trade-offs are outlined to develop win-win strategies. A real case using data analysis tools is analyzed. Results provide guidelines not only to make decisions independently on each stage but also to collaboratively work.The authors acknowledge the support of the project 691249, RUCAPS: "Enhancing and implementing knowledge based ICT solutions within high risk and uncertain conditions for agriculture production systems", funded by the European Union's research and innovation programme under the H2020 Marie Skodowska-Curie Actions.Pérez Perales, D.; Rodríguez-Sánchez, MDLÁ.; Ortiz Bas, Á.; Guyon, C. (2020). Assessing the Impact of Pumpkins Plantation, Harvest and Storage Decisions on a Collaborative Supply Chain with Data Analysis Tools. IFIP Advances in Information and Communication Technology. 598:511-523. https://doi.org/10.1007/978-3-030-62412-5_42S511523598Prima, W.A., Xing, K., Amer, Y.: Collaboration and sustainable agri-food supply chain: a literature review. MATEC, 5802004 (2016)Pérez Perales, D., Alarcón Valero, F., Drummond, C., Ortiz, Á.: Towards a sustainable agri-food supply chain model. the case of LEAF. In: Ortiz, Á., Andrés Romano, C., Poler, R., García-Sabater, J.-P. (eds.) Engineering Digital Transformation. LNMIE, pp. 333–341. Springer, Cham (2019). https://doi.org/10.1007/978-3-319-96005-0_40Esteso, A., Alemany, M.M.E., Ortiz, A.: Conceptual framework for designing agri-food supply chains under uncertainty by mathematical programming models. Int. J. Prod. Res. 56(13), 4418–4446 (2018)Bahrami, R., Khodadadi, M., Piry, S., Hassanpanah, D.: The effects of planting methods and head pruning on seed yield and yield components of medicinal pumpkin (Cucurbita Pepo Subsp. Pepo Convar. Pepo Var. Styriaca) at low temperature areas. Pak. J. Bio. Sci. 12(6), 538–541 (2009)Tarus, W.J., Ochuodho, J.O., Rop, N.K.: Influence of harvesting stage on seed quality aspects of pumpkin (Cucurbita Pepo L.). J. Exp. Agr. 18(2), 1–9 (2017)Amodio, M.L., Rinaldi, R., Colell, G.: Extending shelf life of fresh-cut pumpkin (Cucurbita Maxima): effect of pre-treatments and storage conditions. Acta Hortic. 876, 333–340 (2010)Pimentel, D., et al.: Annual vs perennial grain production. Agric. Ecosyst. Environ. 161, 1–9 (2012)Biesiada, A., Nawirska, A., Kucharska, A., Sokół-Łętowska, A.: The effect of nitrogen fertilization methods on yield and chemical composition of pumpkin (Cucurbita maxima) fruits before and after storage. Veg. Crops. Res. B. 70, 203–211 (2009)Gwenael, J., Paul, Ch., Sau, I., Saurabh, S., Thomasse, S.: Hitting and harvesting pumpkins. Siam. J. Discrete. Math. 28(3), 1363–1390 (2014)Oluoch, M.O.: Production practices of pumpkins for improved productivity. Scripta. Hort. 15, 181–189 (2012)Ondigi, A., Toili, W., Ijani, A., Omuterema, S.: Comparative analysis of production practices and utilization of pumpkins (Cucurbita Pepo and Cucurbita Maxima) by smallholder farmers in the lake victoria basin, East Africa. Afr. J. Environ. Sci. Tech. 2(9), 296–304 (2008)Naik, M.L., Prasad, V.M., Raya, L.P.: A study on character association and path analysis in pumpkin (Cucurbita Moschata Duch. Ex Poir.). Int. J. Adv. Res. 3(1), 1030–1034 (2015)Nerson, H., Paris, H.S., Paris, E.P.: Fruit shape, size and seed yield in Cucurbita Pepo. Acta Hort. 510, 227–230 (2000)Ortiz, A., Alarcón, F., Pérez, D., Alemany, M.M.E.: Identifying the main uncertainties in the agri-food supply chain. In: Mula, J., Barbastefano, R., Díaz-Madroñero, M., Poler, R. (eds.) New global perspectives on industrial engineering and management. LNMIE, pp. 221–229. Springer, Cham (2019). https://doi.org/10.1007/978-3-319-93488-4_25Khalid, E., Mohammed, E.: Dependence of pumpkin yield on plant density and variety. Am. J. Plant. Sci. 2, 636–643 (2011)Jakop, M., et al.: Yield performance and agronomic efficiency in oil pumpkins (Cucurbita Pepo L. Group Pepo) depending on production systems and varieties. Agr. 14(1), 25–36 (2017)Ahmed, B., Masud, M.A.T., Zakaria, M., Hossain, M.M., Mian, M.A.K.: Evaluation of pumpkin (Cucurbita Moschata Duch. Ex Poir.) for yield and other characters. Bangl. J. Agr. Res. 42(1), 1–11 (2017)Zhao, J., et al.: Physicochemical and antioxidant properties of different pumpkin cultivars grown in China. Food Sci. Technol. 9(4), 308–316 (2015)Zhou, C., et al.: The effect of high hydrostatic pressure on the microbiological quality and physical–chemical characteristics of pumpkin (Cucurbita Maxima Duch.) during refrigerated storage. Innov. Food. Sci. Emerg. 21, 24–34 (2014)Loy, J.B.: Harvest period and storage affect biomass partitioning and attributes of eating quality in acorn squash (Cucurbita Pepo), Présenté à Cucurbitaceae, Asheville, North Carolina, USA, pp. 568–577 (2006)Nagao, A., Indou, T., Dohi, H.: Effects of curing condition and storage temperature on postharvest quality of squash fruit. J. Jpn. Soc. Hort. Sci. 60(1), 175–181 (1991)Habibunnisa, R., Prasad, R., Shivaiah, K.M.: Storage behaviour of minimally processed pumpkin (Cucurbita Maxima) under modified atmosphere packaging conditions. Eur. Food Res. Technol. 212(2), 165–169 (2001)Sharrock, K.R., Parkes, S.L.: Physiological changes during development and storage of fruit of buttercup squash in relation to their susceptibility to Rot. New. Zeal. J. Crop. Hort. 18(4), 185–196 (1990)Muzzaffar, S., Baba, W.N., Nazir, N., Masoodi, F.A., Bhat, M.M., Bazaz, R.: Effect of storage on physicochemical, microbial and antioxidant properties of pumpkin (Cucurbita Moschata) candy. Cog. Food. Agr. 2(1), 1163650 (2016)Rahman, M.A., Miaruddin, M., Khan, M.H.H., Masud, M.A.T., Begum, M.M.: Effect of storage periods on postharvest quality of pumpkin. Bangl. J. Agr. Res. 38(2), 247–255 (2013)Song, J., Wei, Q., Wang, X., Li, D., Liu, C., Zhang, M., Meng, L.: Degradation of carotenoids in dehydrated pumpkins as affected by different storage conditions. Food. Res. Int. 107, 130–136 (2018)Matová, A.: the influence of genotype and storage condition on the content of selected bioactive substances in the fruit of pumpkin (Cucurbita Moschata Duch.). J. Microbiol. Bioech. Food. Sci. 9(2), 288–292 (2019)Tilman, D., Balzer, C., Hill, J., Befort, B.L.: Global food demand and the sustainable intensification of agricultura. Proc. Natl. Acad. Sci. U. S. A. 108, 20260–20264 (2011)Wang, W., et al.: Multiple trade‐offs between maximizing yield and minimizing greenhouse gas production in Chinese rice croplands. Land. Degrad. Dev. (2020)Pleijel, H., Uddling, J.: Yield vs. quality trade-offs for wheat in response to carbon dioxide and ozone. Glob. Change. Biol. 18(2), 596–n/a (2011)Pedhazur, E.J., Schmelkin, L.P.: Measurement, Design, and Analysis: an Integrated Approach. Lawrence Erlbaum Associates, Hillsdale (1991

Phase behaviour of Ag2CrO4 under compression: Structural, vibrational, and optical properties

This document is the Accepted Manuscript version of a Published Work that appeared in final form in Journal of Physical Chemistry C, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://dx.doi.org/10.1021/jp401524sWe have performed an experimental study of the crystal structure, lattice dynamics, and optical properties of silver chromate (Ag2CrO4) at ambient temperature and high pressures. In particular, the crystal structure, Raman-active phonons, and electronic band gap have been accurately determined. When the initial orthorhombic Pnma Ag2CrO4 structure (phase I) is compressed up to 4.5 GPa, a previously undetected phase (phase II) has been observed with a 0.95% volume collapse. The structure of phase II can be indexed to a similar orthorhombic cell as phase I, and the transition can be considered to be an isostructural transition. This collapse is mainly due to the drastic contraction of the a axis (1.3%). A second phase transition to phase III occurs at 13 GPa to a structure not yet determined. First-principles calculations have been unable to reproduce the isostructural phase transition, but they propose the stabilization of a spinel-type structure at 11 GPa. This phase is not detected in experiments probably because of the presence of kinetic barriers. Experiments and calculations therefore seem to indicate that a new structural and electronic description is required to model the properties of silver chromate.This study was supported by the Spanish government MEC under grants MAT2010-21270-C04-01/03/04 and CTQ2009-14596-C02-01, by the Comunidad de Madrid and European Social Fund (S2009/PPQ1551 4161893), by the MALTA Consolider Ingenio 2010 project (CSD2007-00045), and by the Vicerrectorado de Investigacion y Desarrollo of the Universidad Politecnica de Valencia (UPV2011-0914 PAID-05-11 and UPV2011-0966 PAID-06-11). A.M. and P.R.-H. acknowledge computing time provided by Red Espanola de Supercomputacion (RES) and MALTA-Cluster. J.A.S. acknowledges Juan de la Cierva Fellowship Program for its financial support. Diamond and ALBA Synchrotron Light Sources are acknowledged for provisions of beam time. We also thank Drs. Peral, Popescu, and Fauth for technical support.Santamaría Pérez, D.; Bandiello, E.; Errandonea, D.; Ruiz-Fuertes, J.; Gomis Hilario, O.; Sans, JÁ.; Manjón Herrera, FJ.... (2013). Phase behaviour of Ag2CrO4 under compression: Structural, vibrational, and optical properties. Journal of Physical Chemistry C. 117(23):12239-12248. https://doi.org/10.1021/jp401524sS12239122481172

Enhancing the sustainability performance of Agri-Food Supply Chains by implementing Industry 4.0

[EN] In order to enhance the sustainability in the supply chain, its members should define and pursue common objectives in the three dimensions of the sustainability (economic, environmental and social). The Agri-Food Supply Chain (AFSC) is a network of different members such as farmers (producers), processors and distributors (wholesales, retailers.), etc.. In order to achieve the performance objectives of the AFSC, Industry 4.0 technologies can be implemented. The aim of this paper is to present a classification of these technologies according to two criteria: objective to be achieved (environmental or social) specified in the main issues to be covered in each objective and member of the AFSC supply chain where it is implemented. In this work, we focus on technologies that deal with environmental and social sustainability because economic sustainability will depend on the specific characteristics of the business (a supply chain using a specific Industry 4.0 technology may be profitable while others do not).This work has been funded by the Project GV/2017/065 "Development of a decision support tool for the management and improvement of sustainability in supply chains" funded by the Regional Government of Valencia. Authors also acknowledge the Project 691249, RUC-APS: Enhancing and implementing Knowledge based ICT solutions within high Risk and Uncertain Conditions for Agriculture Production Systems.Pérez Perales, D.; Verdecho Sáez, MJ.; Alarcón Valero, F. (2019). Enhancing the sustainability performance of Agri-Food Supply Chains by implementing Industry 4.0. IFIP Advances in Information and Communication Technology. 568:496-503. https://doi.org/10.1007/978-3-030-28464-0_43S496503568Camarinha-Matos, L.M., Fornasiero, R., Afsarmanesh, H.: Collaborative networks as a core enabler of Industry 4.0. In: Camarinha-Matos, L.M., Afsarmanesh, H., Fornasiero, R. (eds.) PRO-VE 2017. IAICT, vol. 506, pp. 3–17. Springer, Cham (2017). https://doi.org/10.1007/978-3-319-65151-4_1Stich, V., Gudergan, G., Zeller, V.: Need and solution to transform the manufacturing industry in the age of Industry 4.0 – a capability maturity index approach. In: Camarinha-Matos, L.M., Afsarmanesh, H., Rezgui, Y. (eds.) PRO-VE 2018. IAICT, vol. 534, pp. 33–42. Springer, Cham (2018). https://doi.org/10.1007/978-3-319-99127-6_3Flores, M., Maklin, D., Golob, M., Al-Ashaab, A., Tucci, C.: Awareness towards Industry 4.0: key enablers and applications for internet of things and big data. In: Camarinha-Matos, L.M., Afsarmanesh, H., Rezgui, Y. (eds.) PRO-VE 2018. IAICT, vol. 534, pp. 377–386. Springer, Cham (2018). https://doi.org/10.1007/978-3-319-99127-6_32Seuring, S., Müller, M.: From a literature review to a conceptual framework for sustainable supply chain management. J. Clean. Prod. 16, 1699–1710 (2008)Prima, W.A., Xing, K., Amer, Y.: Collaboration and sustainable agri-food supply chain: a literature review. In: MATEC (2016). https://doi.org/10.1051/matecconf/20165802004Pérez Perales, D., Alarcón Valero, F., Drummond, C., Ortiz, Á.: Towards a sustainable agri-food supply chain model. The case of LEAF. In: Ortiz, Á., Andrés Romano, C., Poler, R., García-Sabater, J.-P. (eds.) Engineering Digital Transformation. LNMIE, pp. 333–341. Springer, Cham (2019). https://doi.org/10.1007/978-3-319-96005-0_40Savastano, M., Amendola, C., Bellini, F., D’Ascenzo, F.: Contextual impacts on industrial processes brought by the digital transformation of manufacturing: a systematic review. Sustainability 11, 891 (2019)Varela, L., Araújo, A., Ávila, P., Castro, H., Putnik, G.: Evaluation of the relation between lean manufacturing, Industry 4.0, and sustainability. Sustainability 11, 1439 (2019)Bonilla, S.H., Silva, H.R.O., da Silva, M.T., Gonçalves, R.F., Sacomano, J.B.: Industry 4.0 and sustainability implications: a scenario-based analysis of the impacts and challenges. Sustainability 10, 3740 (2018)Bányai, T., Tamás, P., Illés, B., Stankeviciute, Z., Bányai, A.: Optimization of municipal waste collection routing: impact of Industry 4.0 technologies on environmental awareness and sustainability. Int. J. Environ. Res. Public Health. 16, 634 (2019)Lin, K.C., Shyu, J.Z., Ding, K.: A cross-strait comparison of innovation policy under Industry 4.0 and sustainability development transition. Sustainability 9, 786 (2017)Kamble, S.: Sustainable Industry 4.0 framework: a systematic literature review identifying the current trends and future perspectives. In: Process Safety and Environmental Protection Transactions of the Institution of Chemical Engineers, Part B, vol. 117, pp. 408–25. Institution of Chemical Engineers (2018)Franciosi, C., Iung, B., Miranda, S., Riemma, S.: Maintenance for sustainability in the Industry 4.0 context: a scoping literature review. IFAC-Pap. Online 51(11), 903–908 (2018)Bocken, N.M.P., Short, S.W., Rana, P., Evans, S.: A literature and practice review to develop sustainable business model archetypes. J. Clean. Prod. 65, 42–56 (2014)Bourlakis, M., Maglaras, G., Aktas, E., Gallear, D., Fotopoulos, C.: Firm size and sustainable performance in food supply chains: insights from Greek SMEs. Int. J. Prod. Econ. 152, 112–130 (2014)Garbie, I.H.: An analytical technique to model and assess sustainable development index in manufacturing enterprises. Int. J. Prod. Res. 52(16), 4876–4915 (2014)Beier, G., Niehoff, S., Ziems, T., Xue, B.: Sustainability aspects of a digitalized industry - a comparative study from China and Germany. Int. J. Precis. Eng. Manuf. Green Technol. 4, 227–234 (2017)Pérez, D., Verdecho, M.J., Alarcón, F: Industry 4.0 for the development of more sustainable decision support tools for agri-food supply chain management. In: 13rd International Conference on Industrial Engineering and Industrial Management, XXIII, Gijón, Spain (2019)Xiaolin, L., Linnan, Y., Lin, P., Wengfeng, L., Limin, Z.: Procedia engineering county soil fertility information management system based on embedded GIS. Procedia Eng. 29, 2388–2392 (2012)Satyanarayana, G.V.: Wireless sensor based remote monitoring system for agriculture using ZigBee and GPS. In: 2013 (CAC2S), pp. 110–114 (2013)Phillips, A.J., Newlands, N.K., Liang, S.H.L., Ellert, B.H.: Integrated sensing of soil moisture at the field-scale: measuring, modeling and sharing for improved agricultural decision support. Comput. Electron. Agric. 107, 73–88 (2014)Liopa-tsakalidi, A., Tsolis, D., Barouchas, P.: Application of mobile technologies through an integrated management system for agricultural production. Procedia Technol. 8, 165–170 (2013). (Haicta)Yerpude, S., Singhal, T.K.: Impact of Internet of Things (IoT) data on demand forecasting. Indian J. Sci. Technol. 10, 5 (2017)Wolfert, S., Ge, L., Verdouw, C., Bogaardt, M.: Big data in smart farming – a review. Agric. Syst. 153, 69–80 (2017)Castka, P., Balzarova, M.A.: ISO 26000 and supply chains-on the diffusion of the social responsibility standard. Int. J. Prod. Econ. 111(2), 274–286 (2008)Stock, T., Obenaus, M., Kunz, S., Kohl, H.: Industry 4.0 as enabler for a sustainable development: A qualitative assessment of its ecological and social potential. Process. Saf. Environ. 118, 254–267 (2018)Verdecho, M.J., Pérez, D., Alarcón F.: Proposal of a customer-oriented sustainable balanced scorecard for agri-food supply chains. In: 12th International Conference on Industrial Engineering and Industrial Management, Girona, Spain, 12–13 July (2018)Valcour, P.M., Hunter, L.W.: Technology, organizations, and work-life integration. In: Kossek, E.E. Lambert, S.J. (eds.), Work and Life Integration: Organizational, Cultural, and Individual Perspectives, pp. 61–84. Lawrence Erlbaum Associates, Mahwah (2005)Arntz, M., Gregory, T., Zierahn, U.: The risk of automation for jobs in OECD countries: a comparative analysis. In: OECD Social, Employment and Migration Working Papers, no. 189. OECD Publishing, Paris (2016)Grubert, J., Langlotz, T., Zollmann, S., Regenbrecht, H.: Towards pervasive augmented reality: context-awareness in augmented reality. IEEE Trans. Vis. Comput. Graph. 23, 1 (2016)Velthuis, A.G.J.: New Approaches to Food-Safety Economics. Kluwer Academic Publishers, Dordrecht (2003)Sándor, Z.P., Csiszár, C.: Development stages of intelligent parking information systems for trucks. Acta Polytechnica Hungarica 10(4), 161–174 (2013)Scognamiglio, V., Arduini, F., Palleschi, G., Rea, G.: Biosensing technology for sustainable food safety. Trends Analyt. Chem. 62, 1–10 (2014)Brynjolfsson, E., McAfee, A.: The Second Machine Age. Work, Progress, and Prosperity in a Time of Brilliant Technologies. W.W. Norton & Company, London (2014)Smith, A., Caiazza, T.: Automation in everyday life (2017). http://assets.pewresearch.org/wpcontent/uploads/sites/14/2017/10/03151500/PI_2017.10.04_Automation_FINAL.pdfHefferon, K.L.: Nutritionally enhanced food crops; progress and perspectives. Int. J. Mol. Sci. 16, 3895–3914 (2015)Glass, S., Fanzo, J.: Genetic modification technology for nutrition and improving diets: an ethical perspective. Curr. Opin. Biotech. 44, 46–51 (2017)Moe, T.: Perspectives on traceability in food manufacture’. Trends Food Sci. Technol. 9(5), 211–214 (1998)Latino, M., Corallo, A., Menegoli, M.: From Industry 4.0 to Agriculture 4.0: how manage product data in agri-food supply chain for voluntary traceability, a framework proposed. In: 20th International Conference on Food and Environment (ICFE), Rome (2018)Linus, U.O.: Traceability in agriculture and food supply chain: a review of basic concepts, technological implications, and future prospects. J. Food Agric. Environ. 1(1), 101–106 (2003)Maumbe, B.M., Okello, J.: Uses of information and communication technology (ICT) in agriculture and rural development in Sub-Saharan Africa: experiences from South Africa and Kenya. IJICTRDA 1(1), 1–22 (2010)Dlodlo, N., Kalezhi, J.: The internet of things in agriculture for sustainable rural development. In: International Conference on Emerging Trends in Networks and Computer Communications (ETNCC) (2015

Adaptive Evolution of the Venom-Targeted vWF Protein in Opossums that Eat Pitvipers

The rapid evolution of venom toxin genes is often explained as the result of a biochemical arms race between venomous animals and their prey. However, it is not clear that an arms race analogy is appropriate in this context because there is no published evidence for rapid evolution in genes that might confer toxin resistance among routinely envenomed species. Here we report such evidence from an unusual predator-prey relationship between opossums (Marsupialia: Didelphidae) and pitvipers (Serpentes: Crotalinae). In particular, we found high ratios of replacement to silent substitutions in the gene encoding von Willebrand Factor (vWF), a venom-targeted hemostatic blood protein, in a clade of opossums known to eat pitvipers and to be resistant to their hemorrhagic venom. Observed amino-acid substitutions in venom-resistant opossums include changes in net charge and hydrophobicity that are hypothesized to weaken the bond between vWF and one of its toxic snake-venom ligands, the C-type lectin-like protein botrocetin. Our results provide the first example of rapid adaptive evolution in any venom-targeted molecule, and they support the notion that an evolutionary arms race might be driving the rapid evolution of snake venoms. However, in the arms race implied by our results, venomous snakes are prey, and their venom has a correspondingly defensive function in addition to its usual trophic role

The Splicing Factor Proline-Glutamine Rich (SFPQ/PSF) Is Involved in Influenza Virus Transcription

The influenza A virus RNA polymerase is a heterotrimeric complex responsible for viral genome transcription and replication in the nucleus of infected cells. We recently carried out a proteomic analysis of purified polymerase expressed in human cells and identified a number of polymerase-associated cellular proteins. Here we characterise the role of one such host factors, SFPQ/PSF, during virus infection. Down-regulation of SFPQ/PSF by silencing with two independent siRNAs reduced the virus yield by 2–5 log in low-multiplicity infections, while the replication of unrelated viruses as VSV or Adenovirus was almost unaffected. As the SFPQ/PSF protein is frequently associated to NonO/p54, we tested the potential implication of the latter in influenza virus replication. However, down-regulation of NonO/p54 by silencing with two independent siRNAs did not affect virus yields. Down-regulation of SFPQ/PSF by siRNA silencing led to a reduction and delay of influenza virus gene expression. Immunofluorescence analyses showed a good correlation between SFPQ/PSF and NP levels in infected cells. Analysis of virus RNA accumulation in silenced cells showed that production of mRNA, cRNA and vRNA is reduced by more than 5-fold but splicing is not affected. Likewise, the accumulation of viral mRNA in cicloheximide-treated cells was reduced by 3-fold. In contrast, down-regulation of SFPQ/PSF in a recombinant virus replicon system indicated that, while the accumulation of viral mRNA is reduced by 5-fold, vRNA levels are slightly increased. In vitro transcription of recombinant RNPs generated in SFPQ/PSF-silenced cells indicated a 4–5-fold reduction in polyadenylation but no alteration in cap snatching. These results indicate that SFPQ/PSF is a host factor essential for influenza virus transcription that increases the efficiency of viral mRNA polyadenylation and open the possibility to develop new antivirals targeting the accumulation of primary transcripts, a very early step during infection

Potential Benefits of Sequential Inhibitor-Mutagen Treatments of RNA Virus Infections

Lethal mutagenesis is an antiviral strategy consisting of virus extinction associated with enhanced mutagenesis. The use of non-mutagenic antiviral inhibitors has faced the problem of selection of inhibitor-resistant virus mutants. Quasispecies dynamics predicts, and clinical results have confirmed, that combination therapy has an advantage over monotherapy to delay or prevent selection of inhibitor-escape mutants. Using ribavirin-mediated mutagenesis of foot-and-mouth disease virus (FMDV), here we show that, contrary to expectations, sequential administration of the antiviral inhibitor guanidine (GU) first, followed by ribavirin, is more effective than combination therapy with the two drugs, or than either drug used individually. Coelectroporation experiments suggest that limited inhibition of replication of interfering mutants by GU may contribute to the benefits of the sequential treatment. In lethal mutagenesis, a sequential inhibitor-mutagen treatment can be more effective than the corresponding combination treatment to drive a virus towards extinction. Such an advantage is also supported by a theoretical model for the evolution of a viral population under the action of increased mutagenesis in the presence of an inhibitor of viral replication. The model suggests that benefits of the sequential treatment are due to the involvement of a mutagenic agent, and to competition for susceptible cells exerted by the mutant spectrum. The results may impact lethal mutagenesis-based protocols, as well as current antiviral therapies involving ribavirin

Characterization in vitro and in vivo of a pandemic H1N1 influenza virus from a fatal case

Pandemic 2009 H1N1 (pH1N1) influenza viruses caused mild symptoms in most infected patients. However, a greater rate of severe disease was observed in healthy young adults and children without co-morbid conditions. Here we tested whether influenza strains displaying differential virulence could be present among circulating pH1N1 viruses. The biological properties and the genotype of viruses isolated from a patient showing mild disease (M) or from a fatal case (F), both without known co-morbid conditions were compared in vitro and in vivo. The F virus presented faster growth kinetics and stronger induction of cytokines than M virus in human alveolar lung epithelial cells. In the murine model in vivo, the F virus showed a stronger morbidity and mortality than M virus. Remarkably, a higher proportion of mice presenting infectious virus in the hearts, was found in F virus-infected animals. Altogether, the data indicate that strains of pH1N1 virus with enhanced pathogenicity circulated during the 2009 pandemic. In addition, examination of chemokine receptor 5 (CCR5) genotype, recently reported as involved in severe influenza virus disease, revealed that the F virus-infected patient was homozygous for the deleted form of CCR5 receptor (CCR5Δ32).Funding Statement: This work was supported by Instituto de Salud Carlos III (Programa especial de investigación sobre la gripe pándemica GR09/0023, GR09/0040, GR09/0039) and Ciber de Enfermedades Respiratorias. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.S

A clinically compatible drug-screening platform based on organotypic cultures identifies vulnerabilities to prevent and treat brain metastasis

We report a medium‐throughput drug‐screening platform (METPlatform) based on organotypic cultures that allows to evaluate inhibitors against metastases growing in situ. By applying this approach to the unmet clinical need of brain metastasis, we identified several vulnerabilities. Among them, a blood–brain barrier permeable HSP90 inhibitor showed high potency against mouse and human brain metastases at clinically relevant stages of the disease, including a novel model of local relapse after neurosurgery. Furthermore, in situ proteomic analysis applied to metastases treated with the chaperone inhibitor uncovered a novel molecular program in brain metastasis, which includes biomarkers of poor prognosis and actionable mechanisms of resistance. Our work validates METPlatform as a potent resource for metastasis research integrating drug‐screening and unbiased omic approaches that is compatible with human samples. Thus, this clinically relevant strategy is aimed to personalize the management of metastatic disease in the brain and elsewhere

Counteracting Quasispecies Adaptability: Extinction of a Ribavirin-Resistant Virus Mutant by an Alternative Mutagenic Treatment

[Background] Lethal mutagenesis, or virus extinction promoted by mutagen-induced elevation of mutation rates of viruses, may meet with the problem of selection of mutagen-resistant variants, as extensively documented for standard, nonmutagenic antiviral inhibitors. Previously, we characterized a mutant of foot-and-mouth disease virus that included in its RNA-dependent RNA polymerase replacement M296I that decreased the sensitivity of the virus to the mutagenic nucleoside analogue ribavirin.[Methodology and Principal Findings] Replacement M296I in the viral polymerase impedes the extinction of the mutant foot-and-mouth disease virus by elevated concentrations of ribavirin. In contrast, wild type virus was extinguished by the same ribavirin treatment and, interestingly, no mutants resistant to ribavirin were selected from the wild type populations. Decreases of infectivity and viral load of the ribavirin-resistant M296I mutant were attained with a combination of the mutagen 5-fluorouracil and the non-mutagenic inhibitor guanidine hydrocloride. However, extinction was achieved with a sequential treatment, first with ribavirin, and then with a minimal dose of 5-fluorouracil in combination with guanidine hydrochloride. Both, wild type and ribavirin-resistant mutant M296I exhibited equal sensitivity to this combination, indicating that replacement M296I in the polymerase did not confer a significant cross-resistance to 5-fluorouracil. We discuss these results in relation to antiviral designs based on lethal mutagenesis[Conclusions] (i) When dominant in the population, a mutation that confers partial resistance to a mutagenic agent can jeopardize virus extinction by elevated doses of the same mutagen. (ii) A wild type virus, subjected to identical high mutagenic treatment, need not select a mutagen-resistant variant, and the population can be extinguished. (iii) Extinction of the mutagen-resistant variant can be achieved by a sequential treatment of a high dose of the same mutagen, followed by a combination of another mutagen with an antiviral inhibitor.Work supported by grants BFU2005-00863, BFU2008-02816/BMC, Proyecto Intramural de Frontera del CSIC 200820FO191, FIPSE 36558/06, and Fundacio´n Ramo´n Areces. CIBERehd is funded by Instituto de Salud Carlos III. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscriptPeer reviewe