Search

The highly infectious disease COVID-19 caused by the Betacoronavirus SARS-CoV-2 poses a severe threat to humanity and demands the redirection of scientific efforts and criteria to organized research projects. The international COVID19-NMR consortium seeks to provide such new approaches by gathering scientific expertise worldwide. In particular, making available viral proteins and RNAs will pave the way to understanding the SARS-CoV-2 molecular components in detail. The research in COVID19-NMR and the resources provided through the consortium are fully disclosed to accelerate access and exploitation. NMR investigations of the viral molecular components are designated to provide the essential basis for further work, including macromolecular interaction studies and high-throughput drug screening. Here, we present the extensive catalog of a holistic SARS-CoV-2 protein preparation approach based on the consortium’s collective efforts. We provide protocols for the large-scale production of more than 80% of all SARS-CoV-2 proteins or essential parts of them. Several of the proteins were produced in more than one laboratory, demonstrating the high interoperability between NMR groups worldwide. For the majority of proteins, we can produce isotope-labeled samples of HSQC-grade. Together with several NMR chemical shift assignments made publicly available on covid19-nmr.com, we here provide highly valuable resources for the production of SARS-CoV-2 proteins in isotope-labeled form., This work was supported by Goethe University (Corona funds), the DFG-funded CRC: “Molecular Principles of RNA-Based Regulation,” DFG infrastructure funds (project numbers: 277478796, 277479031, 392682309, 452632086, 70653611), the state of Hesse (BMRZ), the Fondazione CR Firenze (CERM), and the IWB-EFRE-program 20007375. This project has received funding from the European Union’s Horizon 2020 research and innovation program under Grant Agreement No. 871037. AS is supported by DFG Grant SCHL 2062/2-1 and by the JQYA at Goethe through project number 2019/AS01. Work in the lab of KV was supported by a CoRE grant from the University of New Hampshire. The FLI is a member of the Leibniz Association (WGL) and financially supported by the Federal Government of Germany and the State of Thuringia. Work in the lab of RM was supported by NIH (2R01EY021514) and NSF (DMR-2002837). BN-B was supported by theNSF GRFP.MCwas supported byNIH (R25 GM055246 MBRS IMSD), and MS-P was supported by the HHMI Gilliam Fellowship. Work in the labs of KJ and KT was supported by Latvian Council of Science Grant No. VPP-COVID 2020/1-0014. Work in the UPAT’s lab was supported by the INSPIRED (MIS 5002550) project, which is implemented under the Action “Reinforcement of the Research and Innovation Infrastructure,” funded by the Operational Program “Competitiveness, Entrepreneurship and Innovation” (NSRF 2014–2020) and cofinanced by Greece and the EU (European Regional Development Fund) and the FP7 REGPOT CT-2011- 285950–“SEE-DRUG” project (purchase of UPAT’s 700MHz NMR equipment). Work in the CM-G lab was supported by the Helmholtz society. Work in the lab of ABö was supported by the CNRS, the French National Research Agency (ANR, NMRSCoV2- ORF8), the Fondation de la Recherche Médicale (FRM, NMR-SCoV2-ORF8), and the IR-RMN-THC Fr3050 CNRS. Work in the lab of BM was supported by the Swiss National Science Foundation (Grant number 200020_188711), the Günthard Stiftung für Physikalische Chemie, and the ETH Zurich. Work in the labs of ABö and BM was supported by a common grant from SNF (grant 31CA30_196256). This work was supported by the ETHZurich, the grant ETH40 18 1, and the grant Krebsliga KFS 4903 08 2019. Work in the lab of the IBS Grenoble was supported by the Agence Nationale de Recherche (France) RA-COVID SARS2NUCLEOPROTEIN and European Research Council Advanced Grant DynamicAssemblies. Work in the CA lab was supported by Patto per il Sud della Regione Siciliana–CheMISt grant (CUP G77B17000110001). Part of this work used the platforms of the Grenoble Instruct-ERIC center (ISBG; UMS 3518 CNRS-CEA-UGA-EMBL) within the Grenoble Partnership for Structural Biology (PSB), supported by FRISBI (ANR-10-INBS-05-02) and GRAL, financed within the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE- 0003). Work at the UW-Madison was supported by grant numbers NSF MCB2031269 and NIH/NIAID AI123498. MM is a Ramón y Cajal Fellow of the Spanish AEI-Ministry of Science and Innovation (RYC2019-026574-I), and a “La Caixa” Foundation (ID 100010434) Junior Leader Fellow (LCR/BQ/PR19/11700003). Funded by project COV20/00764 fromthe Carlos III Institute of Health and the SpanishMinistry of Science and Innovation to MMand DVL. VDJ was supported by the Boehringer Ingelheim Fonds. Part of this work used the resources of the Italian Center of Instruct-ERIC at the CERM/ CIRMMP infrastructure, supported by the Italian Ministry for University and Research (FOE funding). CF was supported by the Stiftung Polytechnische Gesellschaft. Work in the lab of JH was supported by NSF (RAPID 2030601) and NIH (R01GM123249).

Non-enzymatic glycation and covalent modification of proteins leads to Advanced Glycation End products (AGEs). AGEs are biomarkers of aging and neurodegenerative disease, and can be induced by impaired neuronal signaling. The objective of this study was to investigate if manipulation of dopamine (DA) in vitro using the model protein, bovine serum albumin (BSA), and in vivo using the model organism Drosophila melanogaster, influences fluorescent AGEs (fAGEs) formation as an indicator of dopamine- induced oxidation events. DA inhibited fAGEs-BSA synthesis in vitro, suggesting an anti-oxidative effect, which was not observed when flies were fed DA. Feeding flies cocaine and methamphetamine led to increased fAGEs formation. Mutants lacking the dopaminergic transporter or the D1-type showed further elevation of fAGEs accumulation, indicating that the long-term perturbation in DA function leads to higher production of fAGEs. To confirm that DA has oxidative properties in vivo, we fed flies antioxidant quercetin (QUE) together with methamphetamine. QUE significantly decreased methamphetamine-induced fAGEs formation suggesting that the perturbation of DA function in vivo leads to increased oxidation. These findings present arguments for the use of fAGEs as a biomarker of DA- associated neurodegenerative changes and for assessment of antioxidant interventions such as QUE treatment.

As complexities of addictive behaviors cannot be fully captured in laboratory studies, scientists use simple addiction- associated phenotypes and measure them inlaboratory animals. Locomotor sensitization, characterized by an increased behavioralresponse to the same dose of the drug, has been extensively used to elucidate thegenetic basis and molecular mechanisms of neuronal plasticity. However, to whatextent it contributes to the development of addiction is not completely clear. Wetested if the development of locomotor sensitization to methamphetamine affectsvoluntary self- administration, and vice versa, in order to investigate how two drug-associated phenotypes influence one another. In our study, we used the geneticallytractable model organism, Drosophila melanogaster, and quantified locomotorsensitization and voluntary self- administration to methamphetamine using behavioraltests that were developed and adapted in our laboratory. We show that flies expressrobust locomotor sensitization to the second dose of volatilized methamphetamine, which significantly lowers preferential self- administration of methamphetamine.Naive flies preferentially self-administer food with methamphetamine over plainfood. Exposing flies to volatilized methamphetamine after voluntary self- administration abolishes locomotor sensitization. We tested period null (per01) mutantflies and showed that they do not develop locomotor sensitization, nor do they showpreferential self-administration of methamphetamine. Our results suggest that theremay be partially overlapping neural circuitry that regulates the expression oflocomotor sensitization and preferential self-administration to methamphetamineand that this circuitry requires a functional per gene.

Industrija proizvodnje etanola stvara nusproizvode koji se mogu koristiti kao hrana za domaće životinje. Ti nusproizvodi su bogati proteinima, energijom i fosforom. Istraživanje je provedeno s ciljem utvrđivanja učinka primjene nusproizvoda u hranidbi tovnih pilića starosti 3 tjedna. Pilići su bili podijeljeni u tri skupine hranjene sa 0,15 i 25% DDGS -a (suhi trop s otopinom) u smjesi. Praćen je dnevni prirast, unos hrane i zdravstveno stanje brojlera od drugog tjedna do kraja tova. Također kod klanja praćen je randman, pH, i boja prsiju. Istraživanje je pokazalo da dodavanje DDGS ne utječe negativno na prirast, povećava konverziju, te pozitivno utječe na spektar boja kože prsiju. Cijena prirasta bila najniža kod skupine hranjene sa 25% DDGS – a. Ako se pravilno koriste, nusproizvodi mogu predstavljati djelomičnu zamjenu za kukuruz i soju u obroku., Industry of ethanol production produces by-products that may be used as feed for domestic animals. These by-products are rich in protein, energy and phosphorus. The study was conducted to investigate the effect of by-products in the diets of broilers age of 3 weeks. Chickens were divided into three groups fed with 0.15 and 25% of DDGS (distiller grains with soluble) mixture. What was monitored is daily gain, feed intake and health of broilers from the second week until the end of the fattening period. Also at slaughter was monitored yield, pH, and color chest. Research has shown that adding DDGS does not adversely affect growth, increase conversion, and positively influence the spectrum of chest color. The price of growth was the lowest in the group fed with 25% DDGS. If properly used, by-products can represent a partial replacement for corn and soy in the diet.

Danas se u peradarskoj proizvodnji traže alternativna krmiva koja mogu svojim sastavom uspješno zamjenjivati tradicionalne žitarice kao glavne energetske komponente krmnih smjesa uz istodobno nižu cijenu, a sve s ciljem što isplativije proizvodnje. Sukladno tome, u peradarskoj se proizvodnji vrlo često koristi kukuruzni DDGS (osušeni destilacijski ostatak s otopinom). Cilj ovoga rada bio je procijeniti učinak upotrebe kukuruznog DDGS-a u krmnim smjesama tovnih pilića na proizvodne pokazatelje, troškove proizvodnje te boju kože. Istraživanje je provedeno na ukupno 90 jednodnevnih pilića provenijencije Ross 308 podijeljenih u tri skupine (kontrolna i dvije pokusne skupine). S četiri tjedna starosti u pokusnu skupinu P1 dodano je 15, 0% kukuruznog DDGS-a, a u P2 skupinu 25, 0% kukuruznog DDGS-a. Istraživanje je pokazalo kako dodatak ispitivanog dodatka u navedenim koncentracijama nema negativnog utjecaja na porast tjelesne mase pilića u promatranom razdoblju tova. Dodatkom opisanog alternativnog krmiva neznatno se povećava utrošak hrane za kg prirasta. Nadalje, dodatkom DDGS-a kao alternativnog krmiva uočene su razlike u vrijednostima boje kože pokusnih skupina pilića (L*, b*) u odnosu na kontrolnu skupinu pilića. Istraživanje je pokazalo kako je cijena koštanja krmnih smjesa pokusnih skupina bila niža od 7-12% u odnosu na cijenu koštanja krmne smjese u kontrolnoj skupini.

U radu su prikazani rezultati kvalitativnog istraživanja u kojem su sudjelovali roditelji djece s Williamsovim sindromom (N=3) i roditelji djece s Buloznom epidermolizom (N=7). Cilj istraživanja bio je dobiti uvid u iskustva roditelja djece koja boluju od navedenih rijetkih bolesti i njihove prijedloge za unaprjeđenje podrške u sustavu. Podaci su prikupljeni metodom polustrukturirarnog intervjua, te su analizirani i obrađeni metodom tematske analize. Rezultati istraživanja pokazali su da se roditelji nakon rođenja djeteta s rijetkom bolešću, suočavaju s neznanjem i neprepoznavanjem dijagnoze, nakon uspostave dijagnoze slijede snažne emocionalne reakcije, a na kraju dolazi do prihvaćanja situacije. Iz rezultata proizlazi da su roditeljima najveći izvor podrške obitelj i prijatelji. Rezultati pokazuju da suživot obitelji s rijetkom bolešću dovodi do emocionalne povezanosti i osjećaja bliskosti među članovima obitelji, ali i suočava obitelj sa raznim izazovima, dovodi do potiskivanja potreba druge djece, potrebe za usklađivanjem uloga i socijalne izolacije. Roditelji upozoravaju na potrebu za većim brojem kvalificiranih stručnjaka kao i na potrebu senzibilizacije i osjetljivosti Vlasti za potrebe roditelja i djece s rijetkom bolešću., This paper describes the results from qualitative research in which parents whose children suffer from Williams syndrome (N=3) and Epidermolysis bullosa (N=7) have participated. Main goal of the research was to gain insight into parents’ experience and suggestions for improvements within the social care system. The method of semi-structured interview was used as the method for gathering the data, and they have been analysed using the thematic analysis. The research has shown that parents deal with misguided and unrecognized diagnosis of their child's illness. Once the diagnosis is made, they deal with strong emotional reactions until the final acceptance of the situation. Also, it has been shown that family and friends are the biggest support in dealing with a rare disease. On the one hand, living with a rare disease strengthens family relations, but on the other brings everyday challenges and social isolation. Parents warn of the need for more qualified experts, as well as the need for sensitization and the government’s awareness when it comes to the needs of parents and children who suffer from rare diseases.

Cilj istraživanja bio je dobiti uvid u iskustvo života i dostupne podrške za djecu koja boluju od rijetkih bolesti WS i BE . Podaci su prikupljeni metodom polustrukturirarnog intervjua, te su analizirani i obrađeni temetskom analizom. Iz rezultata proizlazi da su majkama izvor podrške članovi obitelji i prijatelji. Rijetka dijagnoza suočava roditelje s izazovima usklađivanja životnih uloga, socijalnom izolacijom i nerazumijevanjem okoline. Područje istraživanja dinamike obitelji i kvalitete života djece i roditelja te drugih članova obitelji suočenih s rijetkom bološću treba dodatni znanstveno - stručni angažman.

U radu se iznose zapažanja u vezi sa bogatom zbirkom antičkih spomenika pronađenih na teritoriji današnje budvanske opštine. Riječ je o antičkoj nekropoli u kojoj su otkriveni predmeti iz helenističke i rimske epohe (nakit, oruđe i oružje), zatim mozaici pronađeni na ovoj teritoriji, ostaci starih zidina i građevina koje potiču iz ovog perioda (rimske građevine i „budvanski Akropolj”). Ukazano je na značaj svjedočanstava antičke budvanske prošlosti, uz osvrt na metode i principe čuvanja i prezentovanja ovih spomenika uz kritičku analizu problema otuđenja pomenutog nasljeđa. Budva represents one of the oldest settlements on the Adriatic coast. It was governed by various nations; the largest number of material traces was left by Illyrians, Romans, and Greeks. The interest for the heritage of the above mentioned civilizations has never been particularly noteworthy – the statues from the period were discovered by accident in the middle of the previous century, the artifacts have never been thoroughly explored, the majority of objects was either destroyed or stolen due to careless and unprofessional excavations or even donated to museums in Belgrade, Split, Zagreb, and Cetinje. Nowadays, Budva treasures only a small part of its history, which still does not attract the attention of the local people due to the fact the objects lack an adequate presentation. The most important treasury from this period is an antique necropolis situated near the city walls of the Old Town. The necropolis contained various lavishly decorated jewelry, tools, and weapons from the Hellenistic and the Roman periods. Today, we can see numerous antique mosaics and the foundations of the Old Town walls, the so called “Budva Acropolis”, which is the present day citadel. The preserved artifacts are kept in the City Museum of Budva. However, they are not worthily represented – the local people do not possess enough knowledge about the antique period of their history.

The highly infectious disease COVID-19 caused by the Betacoronavirus SARS-CoV-2 poses a severe threat to humanity and demands the redirection of scientific efforts and criteria to organized research projects. The international COVID19-NMR consortium seeks to provide such new approaches by gathering scientific expertise worldwide. In particular, making available viral proteins and RNAs will pave the way to understanding the SARS-CoV-2 molecular components in detail. The research in COVID19-NMR and the resources provided through the consortium are fully disclosed to accelerate access and exploitation. NMR investigations of the viral molecular components are designated to provide the essential basis for further work, including macromolecular interaction studies and high-throughput drug screening. Here, we present the extensive catalog of a holistic SARS-CoV-2 protein preparation approach based on the consortium’s collective efforts. We provide protocols for the large-scale production of more than 80% of all SARS-CoV-2 proteins or essential parts of them. Several of the proteins were produced in more than one laboratory, demonstrating the high interoperability between NMR groups worldwide. For the majority of proteins, we can produce isotope-labeled samples of HSQC-grade. Together with several NMR chemical shift assignments made publicly available on covid19-nmr.com, we here provide highly valuable resources for the production of SARS-CoV-2 proteins in isotope-labeled form.