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Abstract There has been a surge of interest in covalent inhibitors for protein kinases in recent years. Despite success in oncology, the off-target reactivity of these molecules is still hampering the use of covalent warhead-based strategies. Herein, we disclose the development of precision-guided warheads to mitigate the off-target challenge. These reversible warheads have a complex and cyclic structure with optional chirality center and tailored steric and electronic properties. To validate our proof-of-concept, we modified acrylamide-based covalent inhibitors of c-Jun N-terminal kinases (JNKs). We show that the cyclic warheads have high resilience against off-target thiols. Additionally, the binding affinity, residence time, and even JNK isoform specificity can be fine-tuned by adjusting the substitution pattern or using divergent and orthogonal synthetic elaboration of the warhead. Taken together, the cyclic warheads presented in this study will be a useful tool for medicinal chemists for the deliberate design of safer and functionally fine-tuned covalent inhibitors.

Abstract For mitogen-activated protein kinases (MAPKs) a shallow surface—distinct from the substrate binding pocket—called the D(ocking)-groove governs partner protein binding. Screening of broad range of Michael acceptor compounds identified a double-activated, sterically crowded cyclohexenone moiety as a promising scaffold. We show that compounds bearing this structurally complex chiral warhead are able to target the conserved MAPK D-groove cysteine via reversible covalent modification and interfere with the protein-protein interactions of MAPKs. The electronic and steric properties of the Michael acceptor can be tailored via different substitution patterns. The inversion of the chiral center of the warhead can reroute chemical bond formation with the targeted cysteine towards the neighboring, but less nucleophilic histidine. Compounds bind to the shallow MAPK D-groove with low micromolar affinity in vitro and perturb MAPK signaling networks in the cell. This class of chiral, cyclic and enhanced 3D shaped Michael acceptor scaffolds offers an alternative to conventional ATP-competitive drugs modulating MAPK signaling pathways.

There are numerous new technologies whose implementation in food industry is hampered by the fact that people hesitate to invest in expensive systems which they cannot be sure will work or at least are questionable in terms of a given product. Until recently, preservation by HHP, high hydrostatic pressure, was such a technology, and still is today in some branches of the food industry. Investigations were conducted to answer the question of whether the literature, the laboratory, and the industrial (or at least pilot plant) measurements and results agree with one another. We compared the literature data with two HHP systems which were significantly different in terms of treatment capacity, but their efficiency in killing microbes was studied under the same treatment parameters. Our results show that in nearly all cases only minimal differences exist between the data in the literature and the measurements taken on the two appliances.

Loss of BRCA1 or BRCA2 results in genomic instability; however most studies have focused on the role of these proteins in double-strand break repair. Here the authors coupled cell line genetics and whole genome sequencing to investigate the formation of base substitutions and short indels in BRCA1-deficient cells, revealing a role for translesion DNA synthesis regulated by 53BP1.

The herpesvirus protein ORF45 can bind the two host kinases ERK and RSK but how this affects cell signalling is unclear. Here, the authors show that ORF45 hijacks the ERK-RSK complex by binding to the two kinases with optimized linear motifs and interfering with the host kinase docking systems.

Summary: Mutational signatures defined by single base substitution (SBS) patterns in cancer have elucidated potential mutagenic processes that contribute to malignancy. Two prevalent mutational patterns in human cancers are attributed to the APOBEC3 cytidine deaminase enzymes. Among the seven human APOBEC3 proteins, APOBEC3A is a potent deaminase and proposed driver of cancer mutagenesis. In this study, we prospectively examine genome-wide aberrations by expressing human APOBEC3A in avian DT40 cells. From whole-genome sequencing, we detect hundreds to thousands of base substitutions per genome. The APOBEC3A signature includes widespread cytidine mutations and a unique insertion-deletion (indel) signature consisting largely of cytidine deletions. This multi-dimensional APOBEC3A signature is prevalent in human cancer genomes. Our data further reveal replication-associated mutations, the rate of stem-loop and clustered mutations, and deamination of methylated cytidines. This comprehensive signature of APOBEC3A mutagenesis is a tool for future studies and a potential biomarker for APOBEC3 activity in cancer.

The ATF2 transcription factor is phosphorylated by different mitogen-activated protein (MAP) kinases. Here, the authors show that the functionally distinct MAP kinases JNK and p38 control ATF2 through different binding sites and differential phosphorylation, thereby modulating ATF2’s sensitivity to the JNK and p38 pathways.

O presente trabalho aborda a relação do(a) pesquisador(a) com o campo-tema, desfazendo a sobreposição entre campo e espaço geográfico, para refletir sobre o papel do uso de diários de campo na pesquisa-intervenção. A partir da abordagem enativa da cognição, tomamos a noção de afeto como elemento modulador da atenção do(a) pesquisador(a), que se relaciona com a problemática da pesquisa, produzindo registros de aspectos do cotidiano e de suas implicações no diário, que passam a ser percebidos como parte do campo-tema da pesquisa. Tomamos como objeto de análise diários de campo escritos por pesquisadores(as) em uma pesquisa que buscava acompanhar processos de aprendizagem com um jogo digital de localização. Concluímos que o uso do diário de campo acompanha e participa da produção da atenção do(a) pesquisador(a) na sua inserção no campo-tema, de modo que as memórias, hábitos e a inserção do(a) pesquisador(a) nos contextos cotidianos também compõem a pesquisa, tomada como um fazer político que intervém na realidade.

In this paper we discuss the definition of nonsense and its relation to sense-making in the enactive theory of cognition. We take as a starting point the publication "Enactive Cognition at the Edge of Sense-Making: Making Sense of Non-Sense", organized by Massimiliano Cappuccio and Tom Froese, for being a pioneer work in relation to the theme. The importance of sense-making production for the relationship between perception and action is approached from different aspects. We discuss the central propositions of the theories of autopoiesis and enaction, from which are produced three ways of understanding nonsense from an enactive perspective. In these three approaches, contemporary authors suggest that nonsense is an intermediate or mediating element in processes of sense-making. Finally, we discuss the relationship between nonsense and sense, suggesting an articulation between the sensorimotor and linguistic domains based on a non-antagonistic definition of know-what and know-how.

Translesion DNA synthesis (TLS) is a fundamental damage bypass pathway that utilises specialised polymerases with relaxed template specificity to achieve replication through damaged DNA. Misinsertions by low fidelity TLS polymerases may introduce additional mutations on undamaged DNA near the original lesion site, which we termed collateral mutations. In this study, we used whole genome sequencing datasets of chicken DT40 and several human cell lines to obtain evidence for collateral mutagenesis in higher eukaryotes. We found that cisplatin and UVC radiation frequently induce close mutation pairs within 25 base pairs that consist of an adduct-associated primary and a downstream collateral mutation, and genetically linked their formation to TLS activity involving PCNA ubiquitylation and polymerase κ. PCNA ubiquitylation was also indispensable for close mutation pairs observed amongst spontaneously arising base substitutions in cell lines with disrupted homologous recombination. Collateral mutation pairs were also found in melanoma genomes with evidence of UV exposure. We showed that collateral mutations frequently copy the upstream base, and extracted a base substitution signature that describes collateral mutagenesis in the presented dataset regardless of the primary mutagenic process. Using this mutation signature, we showed that collateral mutagenesis creates approximately 10-20% of non-paired substitutions as well, underscoring the importance of the process.

O presente trabalho propõe uma discussão a respeito da heteronormatividade na escola, considerando o contexto de controvérsias e práticas produzidas entre participantes do Movimento Escola Sem Partido e do movimento de ocupação de escolas ocorrido no Brasil em 2016. Discutimos o último movimento a partir de entrevistas com participantes da ocupação de uma escola pública do município de Canguçu (RS), em momento posterior à sua finalização. O acompanhamento de controvérsias em relação a heteronormatividade na escola é realizado na perspectiva de elaboração de uma prática ética, compreendida como competência e expertise a serem desenvolvidas e aprimoradas de modo incorporado, articulando saberes reflexivos e pré-reflexivos. Para isso, abordamos o tema, indicando a relevância de uma política da narratividade que possibilite ampliar a virtualidade de si e do coletivo escolar.

Abstract Background Homologous recombination (HR) repair deficiency arising from defects in BRCA1 or BRCA2 is associated with characteristic patterns of somatic mutations. In this genetic study, we ask whether inactivating mutations in further genes of the HR pathway or the DNA damage checkpoint also give rise to somatic mutation patterns that can be used for treatment prediction. Results Using whole genome sequencing of an isogenic knockout cell line panel, we find a universal HR deficiency-specific base substitution signature that is similar to COSMIC signature 3. In contrast, we detect different deletion phenotypes corresponding to specific HR mutants. The inactivation of BRCA2 or PALB2 leads to larger deletions, typically with microhomology, when compared to the disruption of BRCA1, RAD51 paralogs, or RAD54. Comparison with the deletion spectrum of Cas9 cut sites suggests that most spontaneously arising genomic deletions are not the consequence of double-strand breaks. Surprisingly, the inactivation of checkpoint kinases ATM and CHK2 has no mutagenic consequences. Analysis of tumor exomes with biallelic inactivating mutations in the investigated genes confirms the validity of the cell line models. We present a comprehensive analysis of sensitivity of the investigated mutants to 13 therapeutic agents for the purpose of correlating genomic mutagenic phenotypes with drug sensitivity. Conclusion Our results suggest that no single genomic mutational class shows perfect correlation with sensitivity to common treatments, but the contribution of COSMIC signature 3 to base substitutions, or a combined measure of different features, may be reasonably good at predicting platinum and PARP inhibitor sensitivity.

Video games have been consolidated as an object of study for several areas of knowledge, creating a heterogeneous research field with negative and positive controversy surrounding the experiences of playing. With its widespread popularity in Brazil starting with the 70's, there is no a generation of adults that grew up playing such digital games. This study aims at approaching the subject of what happens when this generation of players start a family and carry on playing their video-games, now with their children. The methodological strategy chosen is that of interviews with parents that play with their children. The research has had the participation of eight fathers and two mothers. The methodological strategy chosen, as well as the analysis of interview records, followed the systemic approach of family studies and studies of enactive cognition. Such theories base our understanding of the research results. In what regards their decisions as to their children's access to games, empiric knowledge about the experience of playing video game seemed to be more regarded then the declarative-conceptual knowledge that the participants usually get from their cultural backgrounds. It has been observed that practises involving video games can involve interesting ways of family socialization in other social contexts.

Changes in heart rate and heart rate variabilty (HRV) were investigated in untrained (UT; starting their first racing season) and detrained (DT; with 1–3 years of race experience) racehorses before and after 14-week conventional training. HRV was measured at rest over 1 h between 9:00 and 10:00 AM on the usual rest day of the horses. The smallest worthwhile change (SWC) rate was calculated for all HRV parameters. UT horses had significantly higher heart rate compared to DT (P

Numerous anti-cancer drugs perturb thymidylate biosynthesis and lead to genomic uracil incorporation contributing to their antiproliferative effect. Still, it is not yet characterized if uracil incorporations have any positional preference. Here, we aimed to uncover genome-wide alterations in uracil pattern upon drug treatments in human cancer cell line models derived from HCT116. We developed a straightforward U-DNA sequencing method (U-DNA-Seq) that was combined with in situ super-resolution imaging. Using a novel robust analysis pipeline, we found broad regions with elevated probability of uracil occurrence both in treated and non-treated cells. Correlation with chromatin markers and other genomic features shows that non-treated cells possess uracil in the late replicating constitutive heterochromatic regions, while drug treatment induced a shift of incorporated uracil towards segments that are normally more active/functional. Data were corroborated by colocalization studies via dSTORM microscopy. This approach can be applied to study the dynamic spatio-temporal nature of genomic uracil.

Effect of temperament on milk yield, lactation length, physico-chemical properties and somatic cell count of Lacaune ewes were evaluated. The investigation was carried out at a sheep farm in the county of Győr-Moson-Sopron. The temperament of 106 Lacaune ewes was measured by the temperament 5-point-scale test (1=very nervous, 5=very quiet) during milking. Furthermore, 42 ewes were randomly selected from a herd of 106 animals for the analysis of milk composition (fat, protein and lactose), pH, electrical conductivity as well as somatic cell count. It was found that the temperament had a significant effect on lactation length and lactation milk production, lactose, electrical conductivity and somatic cell count. Calm ewes had significantly longer lactation (4 score: 220.7 day; 5 score: 201.4 day) as well as higher milk production (4 score: 207.9 kg; 5 score: 193.3 kg) compared to more temperamental animals (2+3 scores: 166.5 day and 135.5 kg; P

Breast cancer is the most commonly occurring cancer in women and the second most common cancer overall. By the age of 80, the estimated risk for breast cancer for women with germline BRCA1 or BRCA2 mutations is around 80%. Genetically engineered BRCA1-deficient mouse models offer a unique opportunity to study the pathogenesis and therapy of triple negative breast cancer. Here we present a newly established Brca1−/−, p53−/− mouse mammary tumor cell line, designated as CST. CST shows prominent features of BRCA1-mutated triple-negative breast cancers including increased motility, high proliferation rate, genome instability and sensitivity to platinum chemotherapy and PARP inhibitors (olaparib, veliparib, rucaparib and talazoparib). Genomic instability of CST cells was confirmed by whole genome sequencing, which also revealed the presence of COSMIC (Catalogue of Somatic Mutations in Cancer) mutation signatures 3 and 8 associated with homologous recombination (HR) deficiency. In vitro sensitivity of CST cells was tested against 11 chemotherapy agents. Tumors derived from orthotopically injected CST-mCherry cells in FVB-GFP mice showed sensitivity to cisplatin, providing a new model to study the cooperation of BRCA1-KO, mCherry-positive tumor cells and the GFP-expressing stromal compartment in therapy resistance and metastasis formation. In summary, we have established CST cells as a new model recapitulating major characteristics of BRCA1-negative breast cancers.

In his work the Clash of Civilizations S. P. Huntington classified Ukraine as a "cleft country". In our view, the current discord in Ukraine is rooted in the political divisions that have frequently characterised the post-communist countries. In Ukraine, owing to a history of divergent socio-economic development in the various regions, these divisions are strongly regional. The dichotomic socio-economic framework reflects not only ethnic and religious differences but also such factors as urbanisation, economic development, and even natural elements. The resulting political divide in Ukraine may be traced to the dichotomy of its national identity. The dividing line is between east and west, urban and rural, and Russian-speaking and Ukrainian-speaking Ukraine. Politically, it is between "Maidan-Ukraine" and "Anti-Maidan-Ukraine". The divide runs along the Uman'-Kharkiv line between the forest zone and the steppe. In the 18 th century the steppe - which had once been a corridor for the nomadic peoples - became a frontier territory for the Ukrainians and then for Tsarist Russia. The Russian ties of cities in the region were further strengthened by industrialisation in the 19th and 20th centuries and by Russification in the Soviet era. The Soviet-made famine (Holodomor) and the events of World War II (the actions of the UPA) heightened Ukraine's political dichotomy. Since independence Ukraine's political elite has failed to address the problem in an adequate manner. The recent escalation of the conflict is due to a lack of political cohesion in the young state and the gravitational effect of the major powers rather than primarily to some kind of civilizational difference. By accentuating Ukraine's economic and financial difficulties, the global crisis has made the country even more vulnerable.

From the 1990s, extensive research was started on the physiological aspects of individual traits in animals. Previous research has established two extreme (proactive and reactive) coping styles in several animal species, but the means of reactivity with the autonomic nervous system (ANS) activity has not yet been investigated in cattle. The aim of this study was the characterization of cardiac autonomic activity under different conditions in cows with different individual characteristics. For this purpose, we investigated heart rate and ANS-related heart rate variability (HRV) parameters of dairy cows (N = 282) on smaller- and larger-scale farms grouped by (1) temperament and (2) behavioural reactivity to humans (BRH). Animals with high BRH scores were defined as impulsive, while animals with low BRH scores were defined as reserved. Cardiac parameters were calculated for undisturbed lying (baseline) and for milking bouts, the latter with the presence of an unfamiliar person (stressful situation). Sympathetic tone was higher, while vagal activity was lower in temperamental cows than in calm animals during rest both on smaller- and larger-scale farms. During milking, HRV parameters were indicative of a higher sympathetic and a lower vagal activity of temperamental cows as compared to calm ones in farms of both sizes. Basal heart rate did not differ between BRH groups either on smaller- or larger-scale farms. Differences between basal ANS activity of impulsive and reserved cows reflected a higher resting vagal and lower sympathetic activity of reserved animals compared to impulsive ones both on smaller- and larger-scale farms. There was no difference either in heart rate or in HRV parameters between groups during milking neither in smaller- nor in larger-scale farms. These two groupings allowed to draw possible parallels between personality and cardiac autonomic activity during both rest and milking in dairy cows. Heart rate and HRV seem to be useful for characterisation of physiological differences related to temperament and BRH.