The Off-Targets of Clustered Regularly Interspaced Short Palindromic Repeats Gene Editing

Funding: The work of VMB was funded by iNOVA4Health – UIDB/Multi/04462/2020 and UIDP/Multi/04462/2020, a program financially supported by Fundação para a Ciência e Tecnologia (FCT)/Ministério da Educação e Ciência through national funds, and the FCT grant PTDC/BEX-BCM/5900/2014.The repurposing of the CRISPR/Cas bacterial defense system against bacteriophages as simple and flexible molecular tools has revolutionized the field of gene editing. These tools are now widely used in basic research and clinical trials involving human somatic cells. However, a global moratorium on all clinical uses of human germline editing has been proposed because the technology still lacks the required efficacy and safety. Here we focus on the approaches developed since 2013 to decrease the frequency of unwanted mutations (the off-targets) during CRISPR-based gene editing.publishersversionpublishe

The cyclin D1 carboxyl regulatory domain controls the division and differentiation of hematopoietic cells

International audienceAbstractBackgroundThe family of D cyclins has a fundamental role in cell cycle progression, but its members (D1, D2, D3) are believed to have redundant functions. However, there is some evidence that contradicts the notion of mutual redundancy and therefore this concept is still a matter of debate.ResultsOur data show that the cyclin D1 is indispensable for normal hematopoiesis. Indeed, in the absence of D1, either in genetic deficient mice, or after acute ablation by RNA interference, cyclins D2 and D3 are also not expressed preventing hematopoietic cell division and differentiation at its earliest stage. This role does not depend on the cyclin box, but on the carboxyl regulatory domain of D1 coded by exons 4–5, since hematopoietic differentiation is also blocked by the conditional ablation of this region.ConclusionThese results demonstrate that not all functions of individual D cyclins are redundant and highlight a master role of cyclin D1 in hematopoiesis

Spontaneous CO release from Ru(II)(CO)2-protein complexes in aqueous solution, cells, and mice.

We demonstrate that Ru(II)(CO)2-protein complexes, formed by the reaction of the hydrolytic decomposition products of [fac-RuCl(κ(2)-H2NCH2CO2)(CO)3] (CORM-3) with histidine residues exposed on the surface of proteins, spontaneously release CO in aqueous solution, cells, and mice. CO release was detected by mass spectrometry (MS) and confocal microscopy using a CO-responsive turn-on fluorescent probe. These findings support our hypothesis that plasma proteins act as CO carriers after in vivo administration of CORM-3. CO released from a synthetic bovine serum albumin (BSA)-Ru(II)(CO)2 complex leads to downregulation of the cytokines interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-α in cancer cells. Finally, administration of BSA-Ru(II)(CO)2 in mice bearing a colon carcinoma tumor results in enhanced CO accumulation at the tumor. Our data suggest the use of Ru(II)(CO)2-protein complexes as viable alternatives for the safe and spatially controlled delivery of therapeutic CO in vivo.We thank the FCT, the EU, and the EPSRC for funding. G.J.L.B. is a Royal Society University Research Fellow.This is the final published version. It first appeared at http://onlinelibrary.wiley.com/doi/10.1002/anie.201409344/abstract

Communication preferences and perceptions of cancer patient during their first medical oncology appointment

© 2023 The Authors. Psycho-Oncology published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.Background: The first medical oncology appointment serves as a platform for patients to comprehend their diagnosis and prognostic implications of cancer. This study aimed to determine patients' communication preferences during their first medical oncology appointment and to assess the disparities between patients' preferences and perceptions. Methods: A total of 169 cancer patients participated by completing the Communication in First Medical Oncology Appointment Questionnaire (C-FAQ), a two-section questionnaire designed to assess patients' preferences and perceptions regarding Content (information provided and its extent), Facilitation (timing and location of information delivery), and Support (emotional support) during their first medical oncology appointment. A comparative analysis was conducted to assess the variations between preferences and perceptions. Results: Content emerged as the most significant dimension compared to Facilitation and Support. The physician's knowledge, honesty, and ability to provide clear information were considered the most important attributes. Patients evaluated most of their preferences as "very important". Patients' perception of the communication dimensions present during their appointment was below preferences for 11 items, indicating significant discrepancies in clinical practice. Conclusions: Patients highly valued their preferences concerning Content, Facilitation, and Support dimensions of communication. However, patient preferences were more prominently oriented towards the Content dimension. The discrepancies between preferences and perceptions should be viewed as an opportunity for enhancing communication skills through training.info:eu-repo/semantics/publishedVersio

Residues of Citrus sinensis (L.) Osbeck as agents that cause a change in antioxidant defense in plants

This work aimed to verify the allelopathic potential of the extract of;Citrus; seeds, for the purpose of adding a sustainable value to the fruit seeds toward their use as industrial residues. The extract was obtained with a Soxhlet apparatus by using hexane, chloroform, and methanol as solvents. The hexane extract of the;Citrus; seeds primarily consisted of linoleic acid (36.6%), followed by α-linoleic acid (25.3%), oleic acid (17.8%), palmitic acid (9.7%), and estearic acid (3.3%). The analysis results indicate that the oil is similar to those used in the cosmetics and food industries and has an economic value from its industrial application. In addition, the use of the oil causes changes in the oxidant balance, germination, and growth of plants.;Este trabalho teve como objetivo verificar o potencial alelopático do extrato de sementes de ;Citrus;, com o objetivo de agregar valor sustentável para a semente de frutas para a sua utilização como resíduo industrial. O extrato foi obtido com aparelho de Soxhlet, utilizando hexano, clorofórmio e metanol como solventes. O extrato hexânico das sementes de ;Citrus; constituiu principalmente por ácido linoleico (36,6%), seguido por ácido α-linoleico (25,3%), ácido oleico (17,8%), ácido palmítico (9,7%) e ácido esteárico (3,3%). As análises indicam que o óleo é semelhante aos utilizados na indústria de cosméticos e de alimentos, tem valor econômico para a utilização na indústria e seu uso provoca alterações no equilíbrio oxidativo, germinação e crescimento.

Intento flexiexistencialista

Intento / coordenação [de] Mário João Alves Chaves. - Lisboa : Universidade Lusíada, 2022. - ISBN 978-989-640-253-2. - P. 1-184

Diagnóstico da pesca ilegal no Estado do Amapá, Brasil

Este trabalho teve como objetivo diagnosticar a ocorrência da pesca ilegal no Estado do Amapá. Foram analisados registros constantes de 521 Autos de Infração lavrados pelo IBAMA no período de 1995 a 2012 que resultaram na aplicação de 11 advertências e 510 multas, gerando a quantia de R$ 2.458.488,98 e apreensão de 31.939,15 kg de peixes, 2.900 caranguejos, um peixe-boi e quelônios. Houve registro de infrações de pesca em 13 dos 16 municípios amapaenses, com destaque para Oiapoque, Amapá, Calçoene, Santana e Macapá que concentraram 88,67% dos procedimentos. Houve registro de apreensão de 31 espécies de peixes, sendo as mais frequentes o Curimatã, Pescada e Corvina.