The inaugural issue of each SoSyM volume-year is usually written during a time of personal and professional reflection. Over the past 12 months, researchers from all disciplines continued to experience the need for flexibility as the COVID-19 pandemic extended its impact across the globe through many variants. This has forced us all to adapt in many ways such that the idea of “virtual everything” permeates our daily discussions. The software and systems modeling community continued to thrive and exhibited a great degree of flexibility and increased production in the overall research output, despite the limitations for in-person collaborations. In fact, for many years, the first issue of each SoSyM volume-year has reported growth in submissions and other important metrics. The same is true for the 2021 publication year of SoSyM. The journal continues a positive and healthy trajectory in the increased number of submissions and an impact factor that ranks SoSyM among the top publication venues for software engineering.The following sections report on various statistics and general updates about SoSyM that occurred over 2021. We invite all authors to continue submitting their contributions to SoSyM, and we are always available to correspond regarding questions about the suitability of an idea or potential submission. We wish you all of the best for a productive and satisfying year of research and personal growth in 2022!The six SoSyM issues published in 2021 contained 40 Regular papers, 32 Special Section papers, 5 Theme Section papers, 4 Expert Voices, 6 Guest Editorials, and 1 Erratum. In total, 2197 pages were published in volume 20. This is a 38% increase compared to the previous year and continues to represent the commitment by our publisher (Springer) in reducing the time to publication by processing papers expeditiously after acceptance. We are grateful to Elizabeth Dziubela, our Springer liaison, for her helpful efforts in assisting us with the expansion.https://www.sosym.org/.We are excited to report that the 2-year impact factor (IF) for SoSyM remains very respectable at 1.910 (previously at 1.876 in 2020 and 2.66 in the record year of 2019). The 5-year IF increased to 2.074 (from 1.915 last year). Furthermore, the h-5 Google Scholar ranking places SoSyM at #14 among all conferences and journals related to software engineering and programming languages. Further rankings can be found at We have observed that modeling continues to develop in overall maturity across the core domains of Software Engineering and Information Systems. Additionally, there is a growing interest in SoSyM among researchers who focus on model-based/model-driven activities in a wider area of software and systems engineering. This includes man-made systems, such as cars, airplanes, cell phones, and health care devices, but also existing systems from nature, which include biological systems, chemical structures and interactions, and of course complex physical or medical activities in various areas. Thus, it was natural that the number of submissions increased in 2021 over previous years. Over the past year, SoSyM received 420 submissions—more than 50 additional submissions compared to 2020 and the largest number for any year in our history. Also, the number of downloads increased again. At the end of 2020, there were 179,555 downloaded SoSyM articles during the calendar year. Comparatively, the final 2021 yearly download total was 197,730.The acceptance rate increased in 2021 to 34.9% which seems to be coming from the fact that SoSyM has a strong reputation and the number of low-quality papers submitted has considerably decreased. Unfortunately, the average time from submission to the final decision (accept or reject) has also increased to 170 days (146 days in 2020). It remains a challenge for our Editors to identify reviewers amid a community that has the specific expertise in areas covered by SoSyM and the increased submissions are starting to exhibit new challenges in the decision time.When a research community matures, it is often interesting to take a look back through history to observe what contributions had the most impact and what topics emerged as most prominent over specific periods of time. Our collaboration with the MODELS conference has provided an opportunity for us to honor the authors of the most influential papers in our community. Each year, SoSyM identifies the two papers (from the Regular and Theme Section areas) that had the most impact over the past decade since their publication. The selection is based on the ISI citation index among papers published in SoSyM since 2010. The following two papers were presented virtually at MODELS 2021, and each author received an award certificate. We congratulate the authors for these “Most Influential” papers of SoSyM over the past decade.SoSyM 2021 "10-year most influential Regular paper award" was given to:Journal on Software and Systems Modeling (SoSyM), Volume 10, Issue 4, pp. 489–514, Springer, October 2011.Zoltán Micskei and Hélène Waeselynck, "The many meanings of UML 2 Sequence Diagrams: A survey", In: https://doi.org/10.1007/s10270-010-0157-9The SoSyM 2021 "10-year most influential Theme Section paper award" was given to:Journal on Software and Systems Modeling (SoSyM), Volume 10, Issue 3, pp. 313–336, Springer, July 2011.Simona Bernardi, José Merseguer, and Dorina C. Petriu, "A dependability profile within MARTE", In: https://doi.org/10.1007/s10270-009-0128-1More information about the awards can be found at: http://www.sosym.org/awards/.The Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 3, pp. 611–639, Springer, June 2021. https://doi.org/10.1007/s10270-021-00876-zMoussa Amrani, Dominique Blouin, Robert Heinrich, Arend Rensink, Hans Vangheluwe, and Andreas Wortmann, "Multi-paradigm modelling for cyber–physical systems: A descriptive framework", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 3, pp. 685–709, Springer, June 2021. https://doi.org/10.1007/s10270-020-00856-9Sebastian Pilarski, Martin Staniszewski, Matthew Bryan, Frederic Villeneuve, and Dániel Varró, "Predictions-on-chip: model-based training and automated deployment of machine learning models at runtime", In: Journal on Software and Systems Modeling (SoSyM), Springer, in press, 2021. https://doi.org/10.1007/s10270-021-00900-2Ferenc A. Somogyi, Gergely Mezei, Zoltán Theisz, Sándor Bácsi, and Dániel Palatinszky, "Playground for multi-level modeling constructs", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 3, pp. 711–724, Springer, June 2021. https://doi.org/10.1007/s10270-020-00858-7Adrien Le Coënt, Julien Alexandre dit Sandretto, and Alexandre Chapoutot, "Guaranteed master for interval-based cosimulation", In: Journal on Software and Systems Modeling (SoSyM), Springer, in this issue, 2021. https://doi.org/10.1007/s10270-021-00897-8Léa Brunschwig, Esther Guerra, and Juan de Lara, "Modelling on mobile devices—A systematic mapping study", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 6, pp. 2155–2197, Springer, December 2021. https://doi.org/10.1007/s10270-021-00888-9Saša Kuhar and Gregor Polancic, "Conceptualization, measurement, and application of semantic transparency in visual notations—A systematic literature review", In: Journal on Software and Systems Modeling (SoSyM), Springer, in this issue, 2021. https://doi.org/10.1007/s10270-021-00899-6Walter Cazzola, Sudipto Ghosh, Mohammed Al-Refai, and Gabriele Maurina, "Bridging the model-to-code abstraction gap with fuzzy logic in model-based regression test selection", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 5, pp. 1735–1774, Springer, October 2021. https://doi.org/10.1007/s10270-021-00879-wGábor Bergmann, "Controllable and decomposable multidirectional synchronizations", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 4, pp. 1183–1213, Springer, August 2021. https://doi.org/10.1007/s10270-020-00842-1Javier Troya, Nathalie Moreno, Manuel F. Bertoa, and Antonio Vallecillo, "Uncertainty representation in software models: A survey", In: Journal on Software and Systems Modeling (SoSyM), Springer, in this issue, 2021. https://doi.org/10.1007/s10270-021-00905-xLissette Almonte, Esther Guerra, Iván Cantador, and Juan de Lara, "Recommender systems in model-driven engineering—A systematic mapping review", In: Journal on Software and Systems Modeling (SoSyM), Springer, in this issue, 2021. https://doi.org/10.1007/s10270-021-00907-9Martina De Sanctis, Ludovico Iovino, Maria Teresa Rossi, and Manuel Wimmer, "MIKADO: A smart city KPIs assessment modeling framework", In: Journal on Software and Systems Modeling (SoSyM), Springer, in this issue, 2021. https://doi.org/10.1007/s10270-021-00898-7Wenjun Xiong, Emeline Legrand, Oscar Åberg, and Robert Lagerström, "Cyber security threat modeling based on the MITRE Enterprise ATT&CK Matrix", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 6, pp. 2111–2130, Springer, December 2021. https://doi.org/10.1007/s10270-021-00883-0Alexander Boll, Florian Brokhausen, Tiago Amorim, Timo Kehrer, and Andreas Vogelsang, "Characteristics, potentials, and limitations of open-source Simulink projects for empirical research", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 5, pp. 1713–1734, Springer, October 2021. https://doi.org/10.1007/s10270-021-00884-zOszkár Semeráth, Aren A. Babikian, Boqi Chen, Chuning Li, Kristóf Marussy, Gábor Szárnyas, and Dániel Varró, "Automated generation of consistent, diverse and structurally realistic graph models", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 5, pp. 1609–1631, Springer, October 2021. https://doi.org/10.1007/s10270-021-00869-yStéphanie Challita, Fabian Korte, Johannes Erbel, Faiez Zalila, Jens Grabowski, and Philippe Merle, "Model-based cloud resource management with TOSCA and OCCI", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 5, pp. 1553–1586, Springer, October 2021. https://doi.org/10.1007/s10270-021-00868-zShahar Maoz and Jan Oliver Ringert, "Spectra: A specification language for reactive systems", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 4, pp. 1123–1158, Springer, August 2021. https://doi.org/10.1007/s10270-020-00854-xSwaib Dragule, Thorsten Berger, Claudio Menghi, and Patrizio Pelliccione, "A survey on the design space of end-user-oriented languages for specifying robotic missions", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 3, pp. 897–916, Springer, June 2021. https://doi.org/10.1007/s10270-020-00834-1Weslley Torres, Mark G. J. van den Brand, and Alexander Serebrenik, "A systematic literature review of cross-domain model consistency checking by model management tools", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 3, pp. 767–793, Springer, June 2021. https://doi.org/10.1007/s10270-020-00827-0Pablo Gómez-Abajo, Esther Guerra, Juan de Lara, and Mercedes G. Merayo, "Wodel-Test: A model-based framework for language-independent mutation testing", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 5, pp. 1653–1688, Springer, October 2021. https://doi.org/10.1007/s10270-021-00870-5Jaime Font, Lorena Arcega, Øystein Haugen, and Carlos Cetina, "Handling nonconforming individuals in search-based model-driven engineering: nine generic strategies for feature location in the modeling space of the meta-object facility", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 5, pp. 1689–1712, Springer, October 2021. https://doi.org/10.1007/s10270-021-00878-xSina Madani, Dimitris Kolovos, and Richard F. Paige, "Distributed model validation with Epsilon", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 5, pp. 1587–1608, Springer, October 2021. https://doi.org/10.1007/s10270-021-00864-3Maher Fakih, Oliver Klemp, Stefan Puch, and Kim Grüttner, "A modeling methodology for collaborative evaluation of future automotive innovations", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 5, pp. 1491–1523, Springer, October 2021. https://doi.org/10.1007/s10270-020-00853-yMahsa Panahandeh, Mohammad Hamdaqa, Bahman Zamani, and Abdelwahab Hamou-Lhadj, "MUPPIT: A method for using proper patterns in model transformations", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 5, pp. 1525–1551, Springer, October 2021. https://doi.org/10.1007/s10270-020-00855-wSiamak Farshidi, Slinger Jansen, and Sven Fortuin, "Model-driven development platform selection: Four industry case studies", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 4, pp. 1159–1181, Springer, August 2021. https://doi.org/10.1007/s10270-020-00841-2Arvind Nair, Xia Ning, and James H. Hill, "Using recommender systems to improve proactive modeling", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 3, pp. 867–895, Springer, June 2021. https://doi.org/10.1007/s10270-020-00833-2David Granada, Juan M. Vara, Mercedes Merayo, and Esperanza Marcos, "CEViNEdit: Improving the process of creating cognitively effective graphical editors with GMF", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 4, pp. 1043–1077, Springer, August 2021. https://doi.org/10.1007/s10270-020-00839-wMatias Pol'la, Agustina Buccella, and Alejandra Cechich, "Analysis of variability models: A systematic literature review", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 3, pp. 821–835, Springer, June 2021. https://doi.org/10.1007/s10270-020-00830-5Feng Zhu and Jun Tang, "Graphical composite modeling and simulation for multi-aircraft collision avoidance", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 3, pp. 837–866, Springer, June 2021. https://doi.org/10.1007/s10270-020-00831-4Stefan Klikovits and Didier Buchs, "Pragmatic reuse for DSML development", In: Journal on Software and Systems Modeling (SoSyM), Volume 20, Issue 3, pp. 795–819, Springer, June 2021. https://doi.org/10.1007/s10270-020-00829-yMojtaba Bagherzadeh, Karim Jahed, Benoit Combemale, and Juergen Dingel, "Live modeling in the context of state machine models and code generation", In: The collaboration between SoSyM and the MODELS conference continued with the organization of the SoSyM “Journal-First” option. This collaboration enables authors of recent SoSyM papers to present their work across the core conference sessions at MODELS. Through this collaboration, SoSyM authors have the opportunity to reach a broader audience to present their work. This includes research talks that explore more depth through analytical and empirical evidence than found in a typical MODELS conference paper. At MODELS 2021, a record number of “SoSyM First” papers were presented (30 papers). We are very thankful to the MODELS 2021 PC Chairs, Shiva Nejati and Daniel Varro, for their help in the integration of the SoSyM First papers into the general MODELS 2021 schedule. The SoSyM First papers presented at MODELS 2021 were the following :Jon Whittle is pursuing interesting new opportunities in his position as director of Australia’s national science agency, CSIRO Data61. His new focus spans many different disciplines of scientific inquiry. Jon stepped down from the SoSyM Editorial Board this year after nearly 20 years of service to the software and systems modeling community. We are grateful for his assistance and wish him all of the best in his new professional endeavors.Colin Atkinson, Dominik Bork, Erwan Bousse, Travis Breaux, Carlos Cuesta, Martina De Sanctis, Juergen Dingel, Dirk Fahland, Loic Helouet, Nicolas Hili, Jörg Holtmann, Jennifer Horkoff, Thomas Kühne, Akhil Kumar, Xavier Le Pallec, Assaf Marron, Bentley Oakes, Anthony Simons, Thomas Vogel, and Marco Wehrmeister.Below is a list of those who reviewed one or more papers for the journal in the last year. The complete list of reviewers can also be found on our website http://www.sosym.org/people/.Mohamed Abdelrazek, Suraj Ajit, Omar Alam, Ian Alexander, Hessa Alfraihi, Shaukat Ali, Joao Paulo Almeida, Ahmad Salim Al-Sibahi, Sanaa Alwidian, Vasco Amaral, Daniel Amyot, Paolo Arcaini, Wesley K. G. Assuncao, Colin Atkinson, Joanne Atlee, Claudia Ayala, Onder Babur, Omar Badreddin, Mira Balaban, Arosha Bandara, Torsten Bandyszak, Olivier Barais, Mikhail Barash, Luciano Baresi, Ankica Barisic, Konstantinos Barmpis, Jorge Barreiros, Angela Barriga, Thais Batista, Edouard Batot, Dinesh Batra, Steffen Becker, Nelly Bencomo, Luca Berardinelli, Robin Bergenthum, Thorsten Berger, Gabor Bergmann, Ilia Bider, Gordon Blair, Dominique Blouin, Alvine Boaye-Belle, Francis Bordeleau, Dominik Bork, Artur Boronat, Paolo Bottoni, Juan Boubeta-Puig, Johann Bourcier, Erwan Bousse, Jonathan Bowen, Drazen Brdjanin, Travis Breaux, Isabel Brito, Jan Broenink, Jean-Michel Bruel, Davide Brugali, Hugo Bruneliere, Alessio Bucaioni, Antonio Bucchiarone, Robert Buchmann, Thomas Buchmann, Andrea Burattin, Erik Burger, Loli Burgueño, Rimantas Butleris, Cristina Cabanillas, Jordi Cabot, Javier Luis Canovas Izquierdo, Rafael Capilla, Emerson Carneiro de Andrade, Victorio Carvalho, Kārlis Čerāns, Carlos Cetina, Moharram Challenger, Michel Chaudron, Xin Chen, Antonio Cicchetti, Federico Ciccozzi, Robert Clarisó, Tony Clark, Manuel Clavel, Jane Cleland-Huang, Loek Cleophas, Benoit Combemale, Carlo Combi, José Conejero, Carl Corea, Dolors Costal, Jesús Sánchez Cuadrado, Carlos Cuesta, Alcino Cunha, Alberto da Silva, Fabiano Dalpiaz, Andrea D'Ambrogio, Nancy Day, Jose Luis de la Vara, Juan de Lara, Martina De Sanctis, Pierre De Saqui-Sannes, Marne de Vries, Julien DeAntoni, Renzo Degiovanni, Thomas Degueule, Joerg Desel, Byron DeVries, Xavier Devroey, Claudio Di Ciccio, Juri Di Rocco, Davide Di Ruscio, Vasiliki Diamantopoulou, Marcos Didonet Del Fabro, Juergen Dingel, Julio do Prado Leite, Marlon Dumas, Francisco Duran, Johann Eder, Alexander Egyed, Neil Ernst, Bedilia Estrada, Dirk Fahland, Kleinner Farias, Michael Fellmann, João Miguel Fernandes, Eduardo Fernandez, Nicolas Ferry, Robson Fidalgo, Hans-Georg Fill, Xavier Franch, Ulrich Frank, Marc Frappier, Piero Fraternali, Walid Gaaloul, Kelly Garces, Felix Garcia, Luciano Garcia Banuelos, Antonio García-Domínguez, Ning Ge, Sebastien Gerard, Sepideh Ghanavati, Mohamad Gharib, Aditya Ghose, Holger Giese, Christophe Gnaho, Sebastian Goetz, Arda Goknil, Thomas Goldschmidt, Cláudio Gomes, Abel Gómez, Elena Gómez-Martínez, Miguel Goulão, Catarina Gralha, David Granada, Joel Greenyer, Paul Grefen, Georg Grossmann, Eoin Grua, John Grundy, Giancarlo Guizzardi, Renata Guizzardi, Sarra Habchi, Simon Hacks, Irit Hadar, Nabil Hameurlain, Regina Hebig, Monika Heiner, Robert Heinrich, Maritta Heisel, Nicolas Hili, Georg Hinkel, Knut Hinkelmann, Frank Hogrebe, Jörg Holtmann, Stijn Hoppenbrouwers, Jennifer Horkoff, Marianne Huchard, John Hutchinson, Emilio Insfran, Ludovico Iovino, Muhammad Zohaib Iqbal, Fuyuki Ishikawa, Ana Ivanchikj Moroni, Amin Jalali, Dietmar Jannach, Axel Jantsch, Steve Jenkins, Manfred A. Jeusfeld, Mohamed Jmaiel, Paul Johannesson, Reyes Juarez-Ramirez, Nafiseh Kahani, Jānis Kampars, Geylani Kardas, Gabor Karsai, Timo Kehrer, Steven Kelly, Wael Kessentini, Djamel Eddine Khelladi, Marite Kirikova, Alexander Knapp, Shekoufeh Kolahdouz-Rahimi, Dimitrios Kolovos, Dimitris Kolovos, Anil Koyuncu, Lars Michael Kristensen, Thomas Kuehn, Thomas Kühne, Géza Kulcsár, Vinay Kulkarni, Akhil Kumar, Yvan Labiche, Katsiaryna Labunets, Robert Lagerström, Leen Lambers, Elyes Lamine, Yngve Lamo, Kevin Lano, Xavier Le Pallec, Edward Lee, Martti Lehto, Henrik Leopold, Timothy Lethbridge, Nianyu Li, Shuai Li, Tong Li, Grischa Liebel, Crescencio Lima, Igor Linkov, Patricia Lopez, Víctor López-Jaquero, Oscar Luis, Roman Lukyanenko, Mass Soldal Lund, Xiaoxing Ma, Nuno Macedo, Fernando Macías, Paulo Maia, Mahdi Manesh, Beatriz Marín, Assaf Marron, Salvador Martinez, Nicholas Matragkas, Raimundas Matulevicius, Davide Andrea Mauro, Julio Medina, Anna Medve, Claudio Menghi, Giovanni Meroni, Jose Merseguer, Judith Michael, Fredrik Milani, Mark Minas, Miguel Mira da Silva, Pieter Mosterman, Hassan Mountassir, Haralambos Mouratidis, Saad Mubeen, Paula Muñoz, John Mylopoulos, Elisa Yumi Nakagawa, Elena Navarro, Lukas Netz, Bernd Neumayr, Phu Nguyen, Phuong Nguyen, Nan Niu, Arne Nordmann, Bentley Oakes, Ileana Ober, Johnny Öberg, Edson OliveiraJr, Alessandro Oltramari, Xavier Oriol, Michiel Overeem, Richard Freeman Paige, Elda Paja, Liliana Pasquale, Oscar Pastor, Cecile Péraire, Francisca Pérez, Robert Pettit, Alfonso Pierantonio, Joao Pimentel, Monica Pinto, Stephan Poelmans, Geert Poels, Pascal Poizat, Andrea Polini, Saheed Popoola, Pasqualina Potena, Henderik Proper, Truong Ho Quang, Elisa Quintarelli, Ansgar Radermacher, Akshay Rajhans, Jolyta Ralyté, Qusai Ramadan, Nacim Ramdani, Aurora Ramírez, Eric J. Rapos, Alexander Raschke, Gil Regev, Manfred Reichert, Hajo Reijers, Elvinia Riccobene, Jan Oliver Ringert, Erkuden Rios Velasco, Roberto Rodríguez-Echeverría, Michael Rosemann, Pedro Rossel, Ivan Ruchkin, Marcela Ruiz, Adrian Rutle, Mehrdad Saadatmand, Mahsa Sadi, Andrey Sadovykh, Neda Saeedloei, Houari Sahraoui, Rijul Saini, Mattia Salnitri, Leila Samimi, Jesús Sánchez Cuadrado, Kurt Sandkuhl, Stefan Sauer, Clemens Sauerwein, Christian Schilling, David Schmalzing, David Schmelter, Rainer Schmidt, Stefan Schönig, Stefan Schulte, Ulrik Schultz, Christoph Schütz, Ed Seidewitz, Ronny Seiger, Bran Selic, Laura Semini, Sagar Sen, Arik Senderovich, Estefanía Serral Asensio, Mojtaba Shahin, Ramy Shahin, Mohammadreza Sharbaf, Carla Silva, Anthony Simons, Monique Snoeck, Pnina Soffer, Oleg Sokolsky, Hui Song, Wei Song, Jean-Sebastien Sottet, Thomas Springer, Emmanouela Stachtiari, Matthew Stephan, Perdita Stevens, Janis Stirna, Ketil Stølen, Volker Stolz, Daniel Strüber, Arnon Sturm, Allison Sullivan, Yu Sun, Daniel Sundmark, Gerson Sunye, Angelo Susi, Andreas Symeonidis, Eugene Syriani, Gabriele Taentzer, Jérémie Tatibouet, Paul Temple, Ernest Teniente, Thomas Thüm, Matthias Tichy, Ulyana Tikhonova, Massimo Tisi, Juha-Pekka Tolvanen, Victoria Torres, Konstantinos Traganos, Hanh Nhi Tran, Marina Tropmann-Frick, Javier Troya, Christos Tsigkanos, Katja Tuma, Samuel Tweneboah-Koduah, Mark Utting, Antonio Vallecillo, Nick R. T. P. van Beest, Mark van den Brand, Han van der Aa, Wil M.P. van der Aalst, Tijs van der Storm, Jean Vanderdonckt, Irene Vanderfeesten, Hans Vangheluwe, Juan Manuel Vara, Daniel Varro, Eric Verbeek, Eugenio Villar, Thomas Vogel, Johannes von Oswald, Yves Wautelet, Barbara Weber, Thomas Weber, Marco Wehrmeister, Ran Wei, Nils Weidmann, Hans Weigand, Mathias Weske, Bernhard Westfechtel, Martin Weyssow, Manuel Wimmer, Genta Indra Winata, Carson Woo, Andreas Wortmann, Franz Wotawa, Sebastian Wrede, Jianqing Wu, Wenhua Yang, Bahman Zamani, Anna Zamansky, Jelena Zdravkovic, Philipp Zech, Bernard P. Zeigler, Lingfang Zeng, Peng Zeng, Li Zhang, Man Zhang, Nan Zhang, Haiyan Zhao, Athanasios Zolotas, Steffen Zschaler, and Albert Zuendorf.A strong research community depends on the efforts of volunteers who help serve as reviewers. The software and systems modeling community has always risen to the request for help from SoSyM. We appreciate all of the help that the reviewers provided in service to the modeling community! We would also like to offer special recognition to the following reviewers, who were recommended as the SoSyM Best Reviewers of 2021, based on the technical depth and feedback provided to authors over the past year—congratulations! Each of the following reviewers received a certificate of recognition:“Models: The fourth dimension of computer science—Towards studies of models and modelling,” contributed by Bernhard Thalheim. Bernhard is a long-time member of our community, and his paper contains many insights and action points for your consideration. We strongly recommend his paper for your consideration and reflection on the role of modeling in computer science.This issue also includes a special Expert Voice article, with the thought provoking title, Expert voice“Models: The fourth dimension of computer science—Towards studies of models and modelling” by Bernhard ThalheimOverview paper"Model-driven engineering for mobile robotic systems: A systematic mapping study" by Giuseppina Casalaro, Giulio Cattivera, Federico Ciccozzi, Ivano Malavolta, Andreas Wortmann, and Patrizio PelliccioneRegular papers"On the automation-supported derivation of domain-specific UML profiles considering static semantics" by Alexander Kraas"Suggesting model transformation repairs for rule-based languages using a contract-based testing approach" by Roberto Rodríguez-Echeverría, Fernando Macías, Adrian Rutle, and José Conejero"An ontological metamodel for cyber-physical system safety, security, and resilience coengineering" by Georgios Bakirtzis, Tim Sherburne, Stephen Adams, Barry Horowitz, Peter Beling, and Cody Fleming"A generic LSTM neural network architecture to infer heterogeneous model transformations" by Loli Burgueño, Jordi Cabot, Shuai Li, and Sebastien Gerard"Cyber security threat modeling based on the MITRE Enterprise ATT&CK Matrix" by Wenjun Xiong, Emeline Legrand, Oscar Åberg, and Robert Lagerström"Modelling on mobile devices—A systematic mapping study" by Léa Brunschwig, Esther Guerra, and Juan de Lara"Bridging the model-to-code abstraction gap with fuzzy logic in model-based regression test selection" by Walter Cazzola, Sudipto Ghosh, Mohammed Al-Refai, and Gabriele Maurina"Guiding the evolution of product-line configurations" by Michael Nieke, Gabriela Sampaio, Thomas Thüm, Christoph Seidl, Leopoldo Teixeira, and Ina Schaefer"Recommender systems in model-driven engineering—A systematic mapping review" by Lissette Almonte, Esther Guerra, Iván Cantador, and Juan de Lara"MIKADO: A smart city KPIs assessment modeling framework" by Martina De Sanctis, Ludovico Iovino, Maria Rossi, and Manuel Wimmer"A method for transforming knowledge discovery metamodel to ArchiMate models" by Ricardo Pérez-Castillo, Andrea Delgado, Francisco Ruiz, Virginia Bacigalupe, and Mario Piattini"Efficient model similarity estimation with robust hashing" by Salvador Martinez, Sebastien Gerard, and Jordi Cabot"Graphic modeling in Distributed Autonomous and Asynchronous Automata (DA3)" by Wiktor Daszczuk"Enhancing software model encoding for feature location approaches based on machine learning techniques" by Ana Cristina Marcén, Francisca Pérez, Oscar Pastor, and Carlos CetinaThe contents of this issue are as follows:We wish you a Happy New Year with the hope that you enjoy reading the papers in this issue!Huseyin Ergin, Jeff Gray, Bernhard Rumpe, and Martin Schindler.


Corvus cornix) may be aware of their own body size. We set up an experimental design in which the crows had to pass through one of three openings to reach the bait. In the first experiment, we studied whether crows prefer a larger hole if all the three are suitable for passage, and what other predictors influence their choice. In the second experiment, we assessed the ability of the crows to select a single passable hole out of three on the first attempt, even though the area of the former was smaller than that of the other two. The results of the first experiment suggest that when choosing among three passable holes, crows prefer those holes that require less effort from them, e.g., they do not need to crouch or make other additional movements. In the second experiment, three of the five crows reliably more often chose a single passable hole on the first try, despite its smaller size. We believe that these results suggest that hooded crows may be aware of their own body size.Body-awareness is one of the manifestations of self-awareness, expressed in the ability of people and animals to represent their own body physical properties. Relatively little work has been devoted to this phenomenon in comparison with the studies of the ability of self-recognition in the mirror, and most studies have been conducted on mammals and human infants. Crows are known to be “clever” birds, so we investigated whether hooded crows ( Self-awareness is the ability of an individual to distinguish its self from the environment and to separate the “self-entity” from the “other-entity.” In its rudimentary form self-awareness is the ability to become the object of one’s own attention and the capacity to ascribe properties to oneself .There are three main approaches to the research in of self-awareness in animals: (1) mirror self-recognition  solve problems for which these properties are needed. Currently, two variants of such tasks are used. One is used to assess the ability of children, elephants and dogs to operate with the idea that their body has weight and to understand that body weight can be an obstacle to the performance of an experimental task .The second approach allows the researcher to assess the subjects’ knowledge of its own size, that is whether they can correlate it with the size and shape of the hole through which they are supposed to pass . The advIn the work with children , a childIn the work with elephants , the aniA similar technique was used to assess body awareness in 54 dogs . In thisIn addition to body weight awareness, children, and dogs were further studied on their body size awareness, i.e., their ability to correlate their own body size with the size and shape of the hole through which they had to pass was evaluated. In working with children , in ordeWith dog subjects , a diffeThe studies of the features of body-awareness in animals are conducted as an addition to the classic mirror test. The phenomenon is considered as the ability of animals to take into account various characteristics of their own body (such as size and weight) as obstacles to achieving the goal. On the one hand, they complement the data on the specificity of self-awareness in those animals that demonstrate self-recognition in the mirror . On the The main difficulty in studying body-awareness is to exclude the effect of learning. This is achieved either by recruiting a large number of subjects with 1–2 tests on each , or by cThe results of numerous independent studies using different experimental approaches show that corvids achieve cognitive feats that are comparable to those exhibited by primates . CorvidsIt is also important to pay attention to the linkage between ontogeny and cognitive performance of corvids. A 2020 study using the Primate Cognition Test Battery (PCTB) demonstrated that full-blown cognitive skills are found as early as 4 month old ravens and do not change at later ages .Corvids are able to track the goals, perceptions, and knowledges that motivate others’ actions , 2017. DCorvus cornix) may be aware of their own body size.The goal of the current work was to investigate whether hooded crows  were used for these experiments. All birds were housed in the outdoor aviary of the Biology Department of Lomonosov Moscow State University, Russia. The crows were wild-caught and, after treatment in a veterinary hospital due to getting injured in the wild, had been kept for about 1–10 years prior to these experiments. The birds were kept in groups in the open-air enclosure (230 × 350 × 280 cm). All birds were identified by a number on a metal leg ring and had not been used for any other experiments in which they were required to pass through an opening in a wall.A total of six hooded crows  were older than one, but no older than 2 years old, while in the second group one bird, “Malyschka” was 4 years and two birds, “Glaz” and “Schnobel” at least 10 years old. We assumed that the age of an animal might influence the cognitive phenomenon under study. Specifically, in this study we assumed that adult birds would make fewer mistakes in choosing a passable hole.The experimental setup  consisteThe presence/absence, size and shape of the holes could be adjusted with additional plates inserted into the grooves on their sides. Square holes (30 × 30 cm) were made in the middle of the two opposite walls of the arena. To one of them, equipped with a remotely opening door, the “start” cell might be attached (the cage in which the bird was placed after catching played the role of the “start” cell). A “finish” cell with a feeder was placed at the other hole in the wall, into which three flour beetle larvae were placed.When working with the first group of birds, the installation was placed in such a way that the “start” cell was opposite to the door of the room. In this case, to the right of the installation was the wall of the room, and to the left there was a free space along which the experimenter moved during preparation for the test.In order to reduce the influence of the inhomogeneity of the surrounding space when working with the second group of birds, the installation was moved to the far part of the room and fenced off with a white cloth from all sides (from floor to ceiling).The video registration system included two EZVIZ C2W cameras connected to an Ezviz Vault Live CS-X5C-4EU video recorder. Monitoring and control of the recording was carried out using the EZVIZ Studio 2.1.1.0 program. One camera was placed above the center of the arena, and the second was located near the left corner on the side of the “start” cell .The birds were transferred to the experimental room and placed in individual cells of 40 × 40 × 40 cm with a drinker with water and a feeder with three mealworm larvae. The experiment was started at least 15 min later.The experimenter placed a “start” cell with a crow to the opening leading into the arena, which was closed with a door, and left the experimental room. From another room, the experimenter opened the door separating the “start” cell and the arena and observed the behavior of the crow on the monitor screen.The test was considered completed if the bird passed through the hole in the partition and entered the “finish” cell in no more than 5 min. If the bird did not pass through the hole for 5 min, the trial was interrupted. In both cases, the “finish” cell together with the crow was set aside and the next test was carried out with another crow. No more than 9 trials were allocated to each bird per day.If the birds refused to go through the openings, then the proportion of meat in their diet was reduced for 1 or 2 days.Two experiments were carried out, in each of which the birds were first trained to pass through smaller holes and then the test was performed.In the first experiment, we studied whether the birds would prefer to get through a larger hole if the other two were also suitable for passing.Initially, birds from both groups were trained to pass through a single opening in the partition and enter the “finish” cell. The largest hole (30 × 30 cm) was used, the location of which was changed quasi-randomly in each subsequent trial, i.e., the same hole was open for no more than two sequential trials, and each of the holes was open an equal number of times (three times each). Nine tests were carried out with each crow.Then all the birds were taught to pass through a single smaller hole. The whole location was changed quasi-randomly in each subsequent trial. For birds from the first group, a rectangular hole was used, oriented vertically with its long side (10 × 18 cm). Initially, we did not plan to use holes oriented horizontally, therefore we did not use them when training the crows of the first group. Eighteen trials were performed by each of the three birds.For birds from the second group, in half of the trials, rectangular holes were oriented with their long side vertically (10 × 20 cm), and in the other half, horizontally (20 × 10 cm). These two types of conditions alternated quasi-randomly. Twelve trials were performed by each of the three birds.We changed the conditions in the second group believing that the difference in the shape and size of the rectangular holes 10 × 20 cm and 20 × 10 cm would be more noticeable for the crows. In the first group, by the 12th trial, the crows solved the problem in less than 1 min, and then this indicator did not decrease, so in the second group we reduced the number of experimental trials from 18 to 12.The partition had three rectangular holes, one of which was larger in area than the other two. It was either wider when all three holes were oriented with the long side vertically, or higher when all three holes were oriented with the long side vertically. The location of the larger hole was changed quasi-randomly.The first group of birds was first tested in 36 trials , in whicIn each trial, two indicators were recorded: the first approach to the hole and the passage through the hole. The approach was considered a situation when the distance between the tip of the bird’s beak and the hole was no more than 10 cm.The statistical analysis was conducted using Statsoft Statistica, version 10.0.1011.0.2).In total, for all six birds, the presence/absence of a connection between the first approach to one of the holes and the passage through it was assessed. To do this, we compared the empirical distribution of the total number of approaches to holes, followed by passing through the same hole, and the number of approaches after which birds passed through another hole, with a hypothetical uniform distribution (50%/50%) using Pearson’s chi-squared test (χn = 6). The following were used as predictors: group number (1 and 2), position of the opening , area of the opening , hole orientation . The number of passages through the holes was used as the response variable. Two-way interactions between the factors were taken into account . Subject ID was included as a random factor. The effect of the differences between the levels of the factors was determined using Tukey’s post hoc test. The model was also checked for the uniformity of the distribution of errors and the absence of collinearity between predictors.The factorial ANOVA was carried out jointly for birds from both groups , gradually decreasing its size: first, holes 10 × 14 cm were used, then 10 × 12 cm, and finally 10 × 10 cm. The latter were minimally passable to the crows, since their size was comparable to the body size of the birds. With each of the first two holes 9 trials were made. With a hole of the minimum size, from 12 to 21 trials were made, ensuring that the crows passed through it in no more than 5 min.There were three holes in the partition. Test and background trials were alternated . In the The results of 12 test trials were analyzed. In each test trial, two indicators were recorded, the first approach to the hole and the first attempt to enter the hole. The approach was considered a situation when the distance between the tip of the bird’s beak and the hole was no more than 10 cm. An attempt was a penetration in which at least half of the crow’s head was on the opposite side of the hole. This was considered an attempt at a passage, not a passage, since the attempt could be made into an impassable opening. Precisely these indicators were considered and analyzed, since we were interested in the choice of the hole made without feedback from the interaction with the walls of the hole.d-value of 0.20 indicates a small effect size, 0.50 indicates a medium-sized effect, and 0.80 indicates a large effect  was assessed in the total number of first approaches to the hole suitable for passage, as well as attempts to enter it . Since our sample consisted of only 5 birds (one crow was excluded from the experiment—see the explanation in the “Results” section), we calculated the effect size for the number of first approaches to the holes in the trial and the first attempts to pass through the holes. We used Cohen’s d as an effect size measurement [e effect ].2).Further analysis was performed for all five birds in total. To identify the presence/absence of a connection between the first approach to one of the holes and an attempt to pass through it, we compared the empirical distribution of the total number of approaches to the holes, followed by an attempt to pass through the same hole, and the number of approaches after which the bird attempted to pass through another hole, with a hypothetical uniform distribution (50%/50%) using Pearson’s chi-squared test . The following were used as predictors: group number (1/2), hole passability (passable/non-passable), hole position (right/center/left), hole area , orientation of the non-passable opening . The following were used as dependent variables: the number of first approaches to the holes in the trial, the number of first attempts to pass through the holes in the trial. Two-way interactions between the factors were taken into account . Subject ID was included as a random factor. The effect of the differences between the levels of the factors was determined using Tukey’s post hoc test. The model was also checked for the uniformity of the distribution of errors and the absence of collinearity of predictors.Factorial ANOVA was carried out for birds from both groups .The summary of the results of all six birds showed that crows significantly more often pass through the hole they first approached, in 363 out of 432 cases , and its position (right/center/left) were predictors influencing the number of passes.F = 12.076, P < 0.001; F = 25.782, P < 0.001; Tukey’s post hoc test, P < 0.01; F = 25.782, P < 0.001; Tukey’s post hoc test, P < 0.01; F = 25.782, P < 0.001; Tukey’s post hoc test, P < 0.01; Birds more often passed through a larger hole [F = 13.554, P < 0.001; Tukey’s post hoc test, P < 0.05; F = 13.554, P < 0.001; Tukey’s post hoc test, P < 0.01; In cases where the holes differed in width rather than height, these crows more often passed through the central hole [In the second group, one of the crows  refused to leave the “start” cell from the 4th test trial, so from that point on the results of the second group were obtained only for two birds.2 = 37.279; df = 1; P < 0.001). In contrast to the background, in 12 test trials we did not reveal a significant connection between the first approach and the first attempt to enter the same hole: only in 38 cases out of 60 crows entered the hole which they first approached .The summary of the results of all five birds showed that during the 24 background trials, the crows significantly more often passed into the same hole to which they made the first approach, i.e., in 104 cases out of 120  = 30.475, P < 0.001; Tukey’s post hoc test, P < 0.01; The birds more often performed the first approach to a passable hole only if it was higher than the unsuitable one [In test trials, the first attempt to pass through a passable hole did not always result in a pass. Birds often made several attempts to pass through it, and only then did they pass. Accordingly, during one test, crows could make several attempts to pass through different holes. On the other hand, there were cases when crows made several attempts to enter an impassable opening. So, the “Dyatel,” in the sixth trial, approached an unsuitable hole and tried to pass through it 33 times.R = 0.975; B = 0.937; P < 0.005). When the birds approached a hole unsuitable for passing, such a pattern was not revealed .However, regression analysis found that crows significantly more often attempted to enter the hole they approached first if it was suitable for passage . “Rodya” made the first approach to the passable hole and then immediately passed through it only in 6 out of 12 trials .Meanwhile, in some birds during 12 test trials, a significant predominance was revealed of the first approach to a passable hole followed immediately by passing through the hole (without approaches to any other hole). In the first group, two out of three birds (“Joe” and “Dyatel”) a significantly more frequent first approach to the passable hole was followed by passing through it. . “Glaz” made the first approach to the permeable hole and then immediately passed through it only in 5 out of 12 trials .In the second group, one of the two remaining birds (“Schnobel”) significantly more often approached the passable hole first and then passed through it .At the same time, the effect size for the number of the first in trial approaches to the holes for five subjects turned out to be large .Each of the five birds reliably more often made their first attempt to pass through a hole suitable for this: “Joe” and “Rodya”—12/12; d = 1; n = 5; M = 11.4, SD = 0.55).At the same time, the effect size for the number of the first in trial attempts to pass through the holes for five subjects turned out to be large  = 533.488; P < 0.001; Birds significantly more often made the first attempts to pass through holes suitable for this [n = 5, cc = 11, P = 0.278).During 12 test trials, the number of first attempts to enter a hole suitable for passing did not change significantly . In addition, the combined influence of the predictors of the position and orientation of the holes, as well as the position and size of the holes was revealed.In conditions when the holes were equally high and differed only in width, the crows preferred to pass through the central one, i.e., chose the shortest way to the “finish” cell. If the holes were equally wide and differed only in height, then there was no preference for the central one.Avoidance of the left opening was observed in all birds. Perhaps this is because, both in the experiments with the first and in the second group, the experimenter was moving along the left side of the experimental setup during the preparation of the trial and after its completion. Also, this could be due to the fact that one of the cameras was installed on the left side of the experimental setup. It is important to note that the group number (1 or 2) did not predict the choice of the hole for passage. Accordingly, the choice of the hole was not influenced by differences in the organization of experimental trials and the type of arrangement of the experimental setup in the room, neither by differences in the age of the birds.The fact that in both groups the birds preferred to pass through the central hole when the holes differed in width  may signal that it was equally convenient for them to pass through both narrow and high holes as well as through wide and high holes, and the major predictor in this case was the choice of the shortest path. When choosing among holes that differ in height , they preferred to go through the higher one, probably because for this they did not need to squat and make other additional movements. This is consistent with evidence that dogs were slower to approach lower holes .First, it should be noted that, during 24 background trials the birds usually passed through the hole which they approached first. In contrast to background trials, we did not find a significant relationship between the first approach and the first attempt to pass through the same hole during 12 test trials. Only if the hole which the birds first approached was suitable for passage, they made the first attempt to pass through it significantly more often. Although in 12 test trials the birds performed the first approach to the passable hole only if it was higher than the non-passable one.We believe that when approaching the hole, the crows determined whether the hole was passable or not. Accordingly, given that in the background probes the crows significantly more often performed the first approach followed by passage through a suitable hole, the task in the test trial  turned out to be more difficult for birds. During the 12 test trials, not all birds approached the passable hole first. However, “Joe,” “Dyatel,” and “Schnobel” significantly more often approached the passable hole first and then passed through it during the test trials.Despite the fact that only three out of 5 crows reliably more often carried out the first approach and then immediately the first attempt to pass, all crows reliably more often made their first attempt to pass through a hole suitable for passing, even if the first approach in the trial was carried out to the non-passable hole. In contrast to the first experiment, no influence of the hole orientation and its position on the choice was revealed.It is important to note that since the number of first attempts to enter a suitable hole did not change reliably during 12 test trials, we can argue that the choice of a suitable hole for passing was not a result of learning during the test trials of the second experiment. Thus, the method we used for organizing the test trials when they followed two background ones, in which the hole was unsuitable for passage, turned out to be effective.In addition, it should be noted that a greater number of first attempts to pass through the passable holes, as well as a greater number of first approaches to the passable holes followed by passage  was not conditioned by learning to penetrate precisely into holes of exact shape and size (squares 10 × 10 cm) formed during training before Experiment 2. This thesis is substantiated by the fact that in 24 background trials the birds significantly more frequently (in 104 cases out of 120) made the first approach and subsequent passage through the passable hole while in these trials the passable hole was of different shape and size.As mentioned above, the orientation of the holes did not affect the number of first attempts to pass through them. However, as in the first experiment, it influenced the number of first approaches. All crows more often made the first approach to a passable square hole only if it was higher than an unusable rectangular one. Thus, birds were attracted primarily by the higher holes, since passing through them required less effort.One should also consider the possibility that the first approaches and first attempts to pass in 12 test trials were influenced by the training that preceded the test trials in the Experiment 2. In 12 test trials, the birds had to choose between one familiar hole through which they passed earlier, and two unfamiliar holes. This must be taken into account in the organization of further similar experiments on animals in order to exclude the undesirable effect of learning, since the aim of training was passing through the holes in general, and the objective of the study was selecting a passable hole thus demonstrating body awareness.Additionally, one should note that, as in the Experiment 1, in the Experiment 2 the group number (1 or 2) was not a predictor of hole selection for the first approach or first passage. In this study we assumed that adult birds would make fewer mistakes in choosing a passable hole. However, this influence was not detected. This is probably due to the fact that full-blown cognitive skills are formed in corvids by the age of 4 months and do not change significantly at the age of 8, 12, and 16 months . All ourIn general, the results obtained showed that despite the presence of a preference for a larger hole in area, revealed in the first experiment, in the second experiment, taking into account both the first attempts to pass through the hole and the first approaches to the hole followed by passage through the hole, it should be concluded that at least 3 out of 6 crows  were able to choose a hole suitable for passage, even if its area was smaller than that of an unsuitable one.Since we took into account the first in trial approach to a hole and the first attempt to pass through one, we can argue that the birds chose the hole suitable for the passage by comparing its size with ideas about the size of their body, and not due to feedback from touching the walls of the hole or whatever else. Considering the large effect size both for first approaches and first attempts to pass, it can be argued that the hooded crows may develop the ability to aware of their own body size.However, we believe it is necessary to conduct additional studies to determine precisely whether the choice by the crows of a suitably sized hole for passage is the evidence of their body awareness. In particular, as noted above, it is necessary to exclude the undesirable effect of learning, since the aim of training was passing through the holes in general, and the objective of the study was selecting a passable hole thus demonstrating body awareness. In addition, this will reveal the evolutionary diversity of cognitive mechanisms that ensure the solution of problems involving taking into account the size of one’s own body.Elaphe radiata are not able to select a hole suitable for passage on the first try, i.e., by comparing its size with ideas about the size of their body, but they are able to learn this after dozens of trials (For example, using a similar technique, it was found that snakes f trials . Also, sf trials .Our results are consistent with data on the high level of brain development and cognitive abilities of corvids . It is iTo date, signs of body size awareness have been found in children aged 22–26 months  and dogsExpanding the set of tests used and advances in methods, as well as improved coverage of the species studied, can help clarify the cognitive architecture of self-awareness .The original contributions presented in the study are included in the article/The animal study was reviewed and approved by the Biology Department of Lomonosov Moscow State University, Russia.IK: conceptualization, formal analysis, data curation, and project administration. AS, IK, and MS: methodology. EE: software, resources, and visualization. EE and AS: validation. MS, SB, and EE: data collection. IK and AS: writing, original draft preparation. AK: writing, review and editing and supervision. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


In the ongoing coronavirus diseases 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), real-time RT-PCR based diagnostic assays have been used for the detection of infection, but the positive signal of real-time RT-PCR does not necessarily indicate the infectivity of the patient. Due to the unique replication system of the coronavirus, primer/probe sets targeted nucleocapsid (N) and spike (S) protein detect the abundantly synthesized subgenomic RNAs as well as the virus genome, possibly making the assay unsuitable for estimation of the infectivity of the specimen, although it has an advantage for the diagnostic tests. In this study, the primer/probe set targeting the open reading frame 1a (ORF1a) gene was developed to specifically detect viral genomic RNA. Then the relation between the ORF1a signal and infectivity of the clinical specimens was validated by virus isolation using VeroE6 cells, which constitutively express transmembrane protease, serine 2, (VeroE6/TMPRSS2). The analytical sensitivity of developed ORF1a set was similar to that of previously developed N and S sets. Nevertheless, in the assay of the clinical specimen, detection rate of the ORF1a gene was lower than that of the N and S genes. These data indicated that clinical specimens contain a significant amount of subgenomic RNAs. However, as expected, the isolation-succeeded specimen always showed an RT-PCR-positive signal for the ORF1a gene, suggesting ORF1a detection in combination with N and S sets could be a more rational indicator for the possible infectivity of the clinical specimens.  The worldwide outbreak of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), originated in Wuhan, China, in December 2019 [COVID-19 is usually diagnosed based on the detection of the SARS-CoV-2 genome using genetic diagnostic methods, such as real-time RT-PCR assays. This is a highly sensitive and specific method to detect the genomes of infectious agents because the presence of several copies of the targeted region’s sequence are enough to obtain a positive result. Early in the outbreak of SARS-CoV-2, it was reported that three primer/probe sets (targeted open reading frame (ORF1), envelope (E), and nucleocapsid (N) genes) developed for the detection of the SARS-like bat beta-coronavirus could be used in the detection of SARS-CoV-2 . These wGiven the unique replication system of the coronavirus, two strategies have been proposed. One is that the full genome-size negative-strand RNA is transcribed from the genomic positive-strand RNA, and acts as a template for producing subgenomic mRNAs (sgmRNAs) . The othDuring the COVID-19 pandemic, real-time RT-PCR-based diagnostic assays have been used for the detection of SARS-CoV-2 infections; however, correlation between the positive signal of real-time RT-PCR and the possibility of virus spread from patient became a problem since PCR-negative results had been required for discharge ,16,17. FIn this study, to support the understanding of the infectivity of PCR-positive specimen, a primer/probe set targeted ORF1a region with equivalent analytical sensitivity with the NIID-N2 set was developed to increase the possibility of genome RNA detection, and the relation between its positive signal and virus isolation was analyzed. Because the ORF1a region sequences were not contained in canonical sgmRNAs, which accounted for majority of sgmRNA and the DNA, nanoball and nanopore sequencing data showed low coverage for the ORF1a region . Coronaviruses produce sgmRNAs during their replication, and nine to ten major sgmRNAs have been identified for SARS-CoV-2 ,13. As sThen, the performance of the NIID-ORF1a set was evaluated using clinical specimens obtained during national surveillance in Japan. Among 25 nasopharyngeal swabs, 15 were negative and 10 were positive for SARS-CoV-2 by NIID-N2 and NIID-S2 sets . All negTo estimate the infectivity of a specimen, virus isolation can provide a helpful clue. We previously reported that serine protease, TMPRSS2-expressing VeroE6 (VeroE6/TMPRSS2) cells are suitable for the isolation of SARS-CoV-2 . The relp = 0.001 and p < 0.001), indicating the amount of NIID-N2 and NIID-S2 targeted sequences were larger than that of NIID-ORF1a, as described above. In addition, among the 57 specimens, the number of positives were 54, 52, and 44 for NIID-N2, NIID-S2, and NIID-ORF1a, showing a significantly lower positive in NIID-ORF1a (p = 0.01). This supports the hypothesis that N- and S-targeted sets tend to detect abundant sgmRNAs. The range of copy numbers (determined by NIID-N2) for successful isolation was 100 to 800 (average 224.5 copies). The lowest the copy number among the virus-isolated specimen determined using the NIID-ORF1a set was log 1.21 copies per reaction (log 3.14 copies/mL). Not all NIID-ORF1a positive specimens could be isolated, but infectious virus was never isolated from NIID-ORF1a negative specimen. Thus, in combination with N- and S-targeted sets, the real-time RT-PCR positive by ORF1a-targeted set, which likely detects the virus genome, could be a rational indicator for the successful virus isolation from specimens, which showed high Ct or Cp values in the diagnostic real-time RT-PCR test, and possible infectivity of the patients. For negative infectiousness confirmation such as in determination of patient discharge, determining the infectivity of the specimen is often challenging. In a previous study , a Ct ofhttps://www.gisaid.org, accessed on 27 Febuary 2022) in the first half of February 2022 as previously described [p = 0.01, n = 4), the NIID-S2 set could detect 2.5 copies of viral RNA, suggesting the set showed sufficient performance (less than five copies) as a diagnostic assay. In the forward primer of the NIID-ORF1a set, T to G substitution of 18th nucleotide accounted for 11.8% of available sequences. The analytical sensitivity of NIID-ORF1a set was 2.5 . These suggest that the NIID assays still work for the detection of VOCs including Omicron variants.In a previous study, we evaluated the performances of NIID-N2 and NIID-S2 assays for VOCs up to Delta ,26. Vari was 2.5  and thatThe purpose of this study was developing an ORF1a-targeted primer/probe set to detect SARS-CoV-2 RNA and evaluate its relation to possible virus isolation. This study showed the NIID-ORF1a set could be helpful to presume the presence of infectious virus in the specimen by the real-time RT-PCR results in combination with NIID-N2 and NIID-S2 sets. Due to the unique replication system of the coronavirus, the region to be targeted by real-time RT-PCR must be carefully selected. Because the N gene is the most abundant in coronavirus-infected cells ,28, it wWe also note that it is important to understand the difference between Ct and Cp values and understand that it is meaningless to compare Ct values directly in tests conducted using different primer/probe sets, analysis conditions, reagents, instruments, and so forth. This was demonstrated in our study, in which there were two to three differences between the NIID-N2, NIID-S, and NIID-ORF1a sets, but the specimens contained similar virus copy numbers. The online publication from Public Health of England provides a useful explanation of the Ct value . This study also suggested that VeroE6/TMPRSS2 cells might be more highly sensitive then VeroE6 cells for the isolation of VOCs of SARS-CoV-2, especially the current Delta-related variants. Although the isolation and maintenance of SARS-CoV-2 using VeroE6 cells without protease induce a deficiency of the furin-recognition site ,31, whicThe analysis of primer/probe mismatches of the NIID assays used in this study suggested that the NIID assays still can work for the detection of Omicron variant, which was dominant in current epidemiology of SARS-CoV-2 , though In summary, the study suggested that the combination of SARS-CoV-2 detection using the ORF1a primer/probe set, as well as N- and S-targeted sets, and viral isolation using VeroE6/TMPRSS2 cells can facilitate the detection of infectious virus in a specimen. In this study, our NIID assays were used for the evaluation; however, other ORF1a-targeting assays could also be used for the same purpose. VeroE6 cells (American tissue culture collection (ATCC), CRL-1586) and VeroE6 cells that constitutively express transmembrane protease, serine 2 (TMPRSS2), (VeroE6/TMPRSS2), ,36 were Pharyngeal swabs, nasal swabs, and sputum collected in virus-transport medium were obtained during national surveillance tests from February to May in 2020, and stored at −80 °C, at almost same time, indicating the influence by the storing condition was able to be considered almost the same for all. Additional nasopharyngeal and nasal swabs were obtained from Discovery Life Sciences . All swabs were used with the approval of the Research and Ethical Committee for the Use of Human Subjects of the National Institute of Infectious Diseases, Japan . Nucleic acid was extracted using TRIzol LS, TRIzol reagent , QIAamp Viral RNA Mini Kits , and/or the QIAamp 96 Virus QIAcube HT kit (Qiagen) in accordance with the manufacturers’ instructions.http://bioinfo.ut.ee/primer3-0.4.0/, accessed on 27 Febuary 2022) based on the sequence of a Japanese viral isolate  as follows: SARS-CoV-2-NIID-ORF1a-F1, 5′-GGAGCTGGTGGCCATAGTTA-3′; SARS-CoV-2-NIID-ORF1a-R2, 5′-TCAAGAGGGTAGCCATCAGG-3′; SARS-CoV-2-NIID-ORF1a-P1, FAM-5′-GGCGACGAGCTTGGCACTGA-3′-BHQ1. The concentration of the primers and probe for the ORF1a set were as follows: NIID-ORF1a-F1, 500 nM; NIID-ORF1a-R2, 500 nM; NIID-ORF1a-P1, 200 nM. Real-time RT-PCR was performed using QuantiTect Probe RT-PCR kits (Qiagen) and LightCycler systems , which equipped the 2nd derivative maximum method for data calculation. The cycling condition and reaction components were as previously described [n = 3). The absorbance was measured by three times serially diluted RNA and the average was used (n = 3). The copy number adjusted control RNA was diluted with PCR-grade water containing RNase inhibitor (1 U/μL) and yeast RNA (10 ng/mL) as carrier for stable storage at −80 °C. Standard curves were generated by measuring serially diluted control RNAs. In the validation assays, the viral RNA copy number was calculated based on the standard curve obtained with the NIID-N2 set. In this study, the analytical sensitivity was defined as the 50% detectable copy number (per reaction) calculated by the Reed-Müench method (n = 4) and used the definition through the study. The NIID-N2 and NIID-S2 assays were conducted as previously described ,7. Prime2 incubator. After 4 days of incubation, the cytopathic effect (CPE) of each well was recorded. In the spike assay, virus stocks, which was centrifuged supernatant of infected cells, were treated with RNase A  at a concentration of 10 μg/mL for 60 min at 37 °C to exclude miscellaneous RNA other than viral RNA in the virion. The treated viral RNA was then extracted with TRIzol LS reagent, and the copy number was calculated by real-time RT-PCR as described above. The virus stocks were serially diluted based on the calculated copy number using virus-transport medium-containing nasopharyngeal swabs that showed negative for respiratory viruses. The dilutants were inoculated onto VeroE6 and VeroE6/TMPRSS2 cells in 96-well plates. After 90 min of adsorption, cells were washed with PBS and incubated with 1%FBS-DMEM at 37 °C in a 5% CO2 incubator. During 4 days of incubation, wells with a CPE were considered as isolation positive. Prior to the virus isolation, RNA was extracted from the specimens using the QIAamp viral RNA mini kit (Qiagen) and the copy number for each targeted region  was determined by real-time RT-PCR using the control RNA templates as described above. Although the volume depended on the concentration and total volume of the specimen, specimens containing at least 100 copies, determined using the NIID-N2 primer/probe set, were inoculated onto VeroE6/TMPRSS2 cells in 24-well plates. Although the maximum of the specimen volume-inoculated depended on the RNA concentration and the amount, specimens containing 100, 200, 400, 800, and 1600 copies were inoculated in each specimen . After 90 min of adsorption, the cells were washed and then incubated with 1% FBS-DMEM-containing antibiotics  at 37 °C in a 5% COhttps://www.gisaid.org/ accessed on 27 February 2022). The data were collected checking “complete” and “low coverage excl” checkboxes and input “human” in the host field. Each sequence was aligned based on the Wuhan-Hu-1 isolate (GenBank: MN908947) using the multiple alignment program for amino acid or nucleotide sequences  [Omicron variants of SARS-CoV-2 genomic sequences deposited from 1 to 13 February 2020 were collated from EpiCoV in the GISAID database (ebruary) . Mismatcebruary) . p value < 0.05 was considered as significant.Statistical analysis was performed using SigmaPlot version 14.5 . The


The impact of ozonation on physicochemical properties of tapioca was studied and fried peanuts coated with different tapioca were characterized. Different ozone oxidation times  and various pH values  were used for tapioca modification. Tapioca oxidized by ozone for 20 min at pH 7 had higher swelling power (SP), water holding capacity (WHC), oil holding capacity (OHC), and viscosity than the native counterpart (P < 0.05). This coincided with the higher carbonyl and carboxyl contents (P < 0.05). The highest frying expansion (FE) with the lowest hardness was attained for fried peanut coated with tapioca oxidized under the aforementioned condition. Therefore, oxidation of tapioca using ozone under optimal conditions could be a potential means to improve frying expansion as well as the crispiness of the fried coated peanuts.Oxidation of tapioca via ozone oxidation was carried out under different conditions in comparison with H Manihot esculenta) is the potential carbohydrate, apart from corn and rice. Cassava is usually processed into cassava starch, known as tapioca. Tapioca has been used for coating of several fried products. Coated peanuts are popular fried snacks made from peanuts coated with tapioca. The crispness is the most important properties of this fried snack, and it has correlation with the expansion properties of the coating material. Tapioca in its native state has limited expansion and crispiness [Cassava . Therefore, the study aimed to characterize the influence of different ozone oxidation times and pH values on the physicochemical properties of tapioca and properties of fried coated peanuts.CBN and CBX contents of ozone-oxidized tapioca are shown in 2O2 (P < 0.05). It indicted that oxidation process with ozone at this condition was more intensive compared to H2O2.Similarly, different pH condition significantly affected CBX content of ozone-oxidized tapioca. CBX content increased as pH increased up to pH 7, and there was no change at pH 9. This phenomenon suggested that the oxidizing species  were excessive and could mutually destroy themselves by coupling . DecompoP < 0.05). Oxidation process positively affected amylose content of tapioca (P > 0.05). However, the sample oxidized with H2O2 had higher amylose content than ozone-oxidized tapioca (P < 0.05). Sandhu et al. [Amylose content of ozone-oxidized tapioca is shown in u et al.  documentu et al. . Accordiu et al. . Depolymu et al. . Accordiu et al. , depolym2O2 for the exception that the highest SP was obtained for the sample oxidized with ozone for 30 min at pH 9. This was possibly due to degradation of starch granule and the presence of carboxyl content, which could improve hydration ability of starch [SP and solubility of the starches are shown in f starch . The higf starch .2O2-oxidized tapioca, which showed the highest solubility. The higher solubility can be related to the degradation of starch granules. Degradation of starch granules also impairs structure of starch granule as indicated by the improved solubility [Ozone-oxidized tapioca had higher solubility than their native counterpart. A similar result was noticeable with Hlubility ,19. Ther2O2-oxidzed sample (P > 0.05). Generally, the higher WHC is associated with the breaking of the glycosidic bonds in starch granules and the formation of functional groups during oxidation process, which can cause erosion in amorphous region [WHC and OHC are shown in s region . In addis region .P > 0.05). The higher OHC is caused by the erosion of amorphous regions and functional groups formation in starch granule, which is expected to facilitate oil to penetrate into the starch granules during frying [A bit different from WHC, OHC of all oxidized starches were lower than the native one. Lower OHC of oxidized starch attained was because of cross-linking induced by slight oxidation ,11. Among frying .RVA curve of pasting properties of native and ozone-oxidized tapioca in different oxidation times at the selected pH 7 and varying pH conditions of selected 20 min are illustrated in The higher viscosity was possibly mediated by hemiacetal cross-linking between depolymerized starch chain due to low level of oxidants . HemiaceP > 0.05). Nevertheless, the highest peak viscosity, breakdown, setback and final viscosity were achieved at pH 7 (At the same time (20 min), for oxidation by ozone, no changes in pasting temperature were noticeable  showed the highest FE (P < 0.05) compared to the others. FE occurred, mostly caused by the vapor pressure of trapped water, which is increased when the temperature is augmented, thus inducing bubble growth. The more water trapped can more likely enhance the more bubble growth in the dough, therefore resulting in higher expansion [Depolymerization of chains in starch granules due to oxidation can increase frying expansion due to the looser structure. FE property of fried coated peanut is depicted in xpansion  reportedTrapped water could be related to hydrophilic ability of the starch granule, in which CBX groups and degradation of the starch granule played a profound role, whereas hydrophilic ability of the starch granule is associated with amylose molecules, which is the major component of the amorphous region ,18. The 2O2-oxidized tapioca in fried peanut. However, the lower hardness (P < 0.05) was found with the oxidation time of 10 and 20 min at pH 7. Nurul et al. [Hardness of fried coated peanut is shown in l et al.  reportedl et al.  suggeste2O2. Porous structure is associated with the crispy texture. The product with smaller pores had higher hardness value. Porosity of product can be examined by the expansion ratio that is in accordance with frying expansion characteristics [The microscopic structures of fried peanuts coated with tapioca are illustrated in eristics .During frying, the water vapor pressure will induce the formation of porous structures in the product . TapiocaNative cassava starch/tapioca and unshelled peanuts (the medium size of average diameter of 7 mm) were procured from the local market in the city of Yogyakarta, Indonesia. All the reagents were of analytical grade.w/v) and the mixture were adjusted to pH 5, 7, and 9 with 0.01 N NaOH or 0.05% citric acid. The suspension was bubbled with ozone (0.18–0.41 g ozone/h) for 10, 20, and 30 min. The suspension was washed until neutral pH was obtained. The oxidized tapioca was then dried at 50 °C for 24 h using an oven dryer until the moisture content of 11% was gained. Ozone-oxidized tapioca was then sieved with 80 mesh sieve. Tapioca oxidized with H2O2 was also used as comparison. Oxidized tapioca with H2O2 was prepared by mixing tapioca suspension with 0.1% H2O2 at pH 7 for 20 min. All tapioca samples including native tapioca were subjected to analyses.Ozone-oxidized tapioca was prepared as tailored by Satmalawati  with a sVb is mL of HCl used for the blank; Vs is mL of HCl used for sample; N is HCl concentration (N), and W is sample weight .CBN content was measured as detailed by Kuakpetoon and Wang . In a toVb is mL of NaOH used for the blank; Vs is mL of NaOH for sample; N is NaO concentration; and W is sample weight .CBX content was measured as reported by Sangseethong et al.  with a sAmylose content was measured according to the method of AOAC . The amyg, 15 min). The supernatant (5 mL) was dried at 110 °C using an oven dryer  until the constant weight was achieved, representing starch solubilized in water. Solubility was expressed as g per 100 g of starch on a dry weight basis.Swelling power and solubility were determined following the method of Adebowale et al. . A starcW2). Swelling power content was calculated as follows:The pellet obtained after centrifugation was transferred to the clean dried test tube and weighed  and the supernatant was discarded. The tube and the pellet were weighed (W2). WHC and OHC were calculated as follows:m. oiled is W2 minus tube weight.WHC and OHC were examined as per the method of Uzomah and Ibe . Water oPasting properties were determined using Rapid Visco Analyzer . To prepare the sample, 25 mL of distilled water were added into 3 g of sample in a RVA canister and mixed thoroughly. Suspension was subjected to heating at 50 °C for 1 min and heating was continued up to 90 °C for 7 min and 30 s with a rate 5.3 °C/min and hold for 5 min. Temperature was then decreased to reach 50 °C with a rate 5.3 °C/min and hold for 2 min. Pasting curve was then recorded and evaluated for the pasting parameters.2O2-oxidized, and ozone-oxidized tapioca starches (5 g) were dispersed and heated in 20 mL of distilled water until gelatinization took place. Gel was then mixed with 17.5 g of dry tapioca (native or oxidized tapioca). The dough was then used to coat unshelled ground peanut with thickness of 2 mm and was subsequently fried in palm oil at 170 °C for 15 min. The products obtained were subjected to analyses.The method of Suprapti  was follV1). After frying, samples were placed again in a measuring glass and the final volume was recorded (V2). FE was calculated as follows:FE of fried coated peanut was determined following the method of Nguyen et al. . Coated Hardness of fried coated peanut was examined as detailed by Nguyen et al. . HardnesMicrostructures of fried coated peanut were visualized as per the method of Saeleaw and Schleining  using a Duncan Multiple’s Range Test (DMRT) at 95% confidence level using SPSS software version 17.0 .The data from each treatment were subjected to analysis of variance (ANOVA) and the significant differences among means were determined by Oxidation process using ozone for different oxidation times and pH conditions significantly affected physicochemical properties of tapioca. SP, viscosity, WHC, and FE increased due to oxidation by ozone, and optimum condition was found at pH 7 for 20 min. This was concomitant with the increased CBN and CBX contents, which were associated with crispy texture or less hardness of fried coated peanut. Tapioca oxidized by ozone under the optimal condition yielded the highest frying expansion of fried coated peanut. It could be related to hydration ability of starch granule, in which CBX groups and degradation of starch granule were augmented.


Academic journalism is akin to building a puzzle in many respects. Every piece has a specific position and role to play and development occurs as the pieces interlock succinctly and mould into something recognisable.Being openly accessible, published manuscripts are permanently under the scrutiny of the scholarly community microscope and therefore sound methodology, ethics, integrity and accuracy are vital components of literary journals. These pieces connect through four major cornerstones, namely: the authors, reviewers, editors and publishers. The interplay between these key role players is crucial in creating a solid framework on which to expand. With the foundation in place, growth can commence; but like a picturesque puzzle, the challenge lies in finding the correct place for all the elements. Occasionally, pieces do not fit compactly and this can be likened to oversights and differing opinions. Whilst these are not always welcome, they do indicate a positive interest in the published material and promote healthy constructive criticism.Where do journal metrics fit? Research journals are continually measured, compared and ranked. These numerical ratings provide a locus for the journal which can change depending on the journal’s performance. The more commonly known metric is the journal impact factor, but others include the CiteScore, the SCImago Journal Rank (SJR), the Eigen factor and the Source Normalised Impact per paper (SNIP). The number of readers, downloads, citations and even popularity on social media, all affect these metrics and ultimately the journal’s prestige.Closely linked to journal metrics is indexing, which involves a formal application for inclusion on a bibliographic or literature database, often linked to an affiliated institution. Selecting the appropriate database for one’s journal is cardinal and inclusion depends on whether all the desired criteria are met. The process of indexing is similar to the peer review process that manuscripts undergo, except that the entire journal undergoes a technical review by an external advisory committee over a certain period, usually 2 years. Ultimately, indexing is all about journal exposure.Similar to matching puzzle pieces, author metrics are aligned with journal metrics and databases. Authors are graded according to their publication numbers, citations and first author publications, amongst other criteria. Most databases will provide authors with a profile calculation, such as the h-index, which allows them to gauge themselves against colleagues. An alternative is to consider author esteem and expertise. A broad range of influential editorial board members and collaborators are helpful in this regard. These pieces are like bridges between the focal point of the puzzle and the background. Additionally, like a visually stimulating puzzle image, journal content should spark interest and relevance, capturing the reader’s attention.South African Journal of Radiology puzzle is intricate and diverse and made up of several elements. Although expanding the journal is an onerous task, all puzzles can be solved and the pieces are finding their places with time.South Africa is a niche geographic location in Africa, and there is a wealth of information from our local disease burden that is unique to our country and worthy of dissemination. The Maya Patel


Athelia (Fibularhizoctonia) sp. strain TMB , which forms termite-egg-mimicking sclerotia for which termites care. We further compare its repertoire of psilocybin gene homologs to homologs previously reported for Fibularhizoctonia psychrophila.Atheliales is a diverse order of crust-forming Basidiomycota fungi. Here, we report the draft genome of the “cuckoo fungus,”  Athelia (Fibularhizoctonia) sp. TMB strain TB5 , which forms termite-egg-mimicking sclerotia that termites tend. We further compare its repertoire of psilocybin gene homologs to homologs previously reported for Fibularhizoctonia psychrophila.Atheliales is a diverse order of crust-forming Basidiomycota fungi. Here, we report the draft genome of the “cuckoo fungus,”  Fibularhizoctonia psychrophila CBS 109695, associated with carrot spoilage  psilocybin gene cluster  is an ecologically diverse –4 order –spoilage , 5, and Athelia (Fibularhizoctonia) sp. TMB  produces sclerotia  that chemically and structurally mimic Reticulitermes sp. eggs and receive care in over 70% of colonies of some Reticulitermes species . Sclerotia were cultured in potato dextrose broth at room temperature for 14 days with shaking. Tissue was filtered through Miracloth, flash-frozen with liquid nitrogen, and pulverized with a mortar and pestle. Genomic DNA was immediately extracted using the DNeasy plant minikit (Qiagen). Short-read DNA libraries were prepared using the NEBNext Ultra DNA library preparation kit and were sequenced with a 150-bp paired-end format on a NovaSeq 6000 system (Illumina). Long reads were generated by fragmenting genomic DNA with a Covaris g-TUBE, preparing libraries using the SQK-LSK108 ligation sequencing kit, and sequencing the libraries on a MinION system (Oxford Nanopore Technologies) using an R9.4 flow cell. Sequencing generated 62,281,267 Illumina reads with an average coverage of 230.98× and 228,721 MinION reads with a mean length of 3,983.38 bp, a median length of 1,834 bp, and a mean coverage of 10.94×. Coverage was calculated by mapping reads to the assembly with Bowtie 2 v2.4.1-2  data set (F. psychrophila expressed sequence tag (EST) and transcript data , Coniophora olivacea MUCL 20566 (https://mycocosm.jgi.doe.gov/Conol1), Coniophora puteana (https://mycocosm.jgi.doe.gov/Conpu1), Paxillus involutus ATCC 200175 (https://mycocosm.jgi.doe.gov/Paxin1), Pisolithus microcarpus 441 (https://mycocosm.jgi.doe.gov/Pismi1), Pisolithus tinctorius Marx 270 (https://mycocosm.jgi.doe.gov/Pisti1), Rhizopogon vesiculosus Smith (https://mycocosm.jgi.doe.gov/Rhives1), Scleroderma citrinum Foug A (https://mycocosm.jgi.doe.gov/Sclci1), Serpula himantioides MUCL 38935 (https://mycocosm.jgi.doe.gov/Serla_varsha1), and Suillus brevipes Sb2 v2.0 (https://mycocosm.jgi.doe.gov/Suibr2). Gene prediction was finalized using OrthoFiller v1.1.1 , C. olivacea MUCL 20566, F. psychrophila CBS 109695, Galerina marginata , Laccaria bicolor v2.0 (https://mycocosm.jgi.doe.gov/Lacbi2), P. croceum F 1598, Plicaturopsis crispa (https://mycocosm.jgi.doe.gov/Plicr1/Plicr1.home.html), Psilocybe serbica (https://mycocosm.jgi.doe.gov/Psiser1), and S. brevipes Sb2 v2.0.Illumina reads were trimmed using Trimmomatic v0.36  with the v1.0.11  and the  v1.0.11 . Genes w v1.0.11 , AUGUSTU v1.0.11 , Glimmer v1.0.11 , and Gen v1.0.11  via the  v1.0.11 . We refedata set , F. psycipt data , and pror v1.1.1  with defF. psychrophila and Athelia sp. TMB TB5 was calculated from the BLASTp . Protein domains were aligned (MAFFT v7.467 [Average amino acid identity (AAI) between e BLASTp  identitye BLASTp  with P. equences  were obtequences  domains T v7.467  with --aT v7.467 , -automaT v7.467 ). Close Athelia sp. TMB TB5 has a mean AAI of 82.58%  with F. psychrophila across 1,727 single-copy orthologs, consistent with substantial divergence time. Multiple copies of psilocybin decarboxylase, hydroxylase, and kinase in Athelia sp. TMB TB5 , and SRR12880627 (Illumina read library).The assembly and annotation of


Hospital Universitario de Álava, Álava, SpainCIBERSAM, Biomedical Research Centre in Mental Health Network, Madrid, SpainUniversidad del País Vasco, Álava, SpainIIS Bioaraba, Álava, SpainThis article when published failed to include the full list of Dr Ana González-Pinto’s affiliations, which are as follows: