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Nucleic Acids Research Sep 2023In severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the non-structural protein NSP1 inhibits translation of host mRNAs by binding to the mRNA entry channel...
In severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the non-structural protein NSP1 inhibits translation of host mRNAs by binding to the mRNA entry channel of the ribosome and, together with the 5'-untranslated region (UTR) of the viral mRNAs, allows the evasion of that inhibition. Here, we show that NSP1 mediates endonucleolytic cleavages of both host and viral mRNAs in the 5'UTR, but with different cleavage patterns. The first pattern is observed in host mRNAs with cleavages interspersed regularly and close to the 5' cap (6-11 nt downstream of the cap). Those cleavage positions depend more on the position relative to the 5' cap than on the sequence itself. The second cleavage pattern occurs at high NSP1 concentrations and only in SARS-CoV-2 RNAs, with the cleavages clustered at positions 45, 46 and 49. Both patterns of cleavage occur with the mRNA and NSP1 bound to the ribosome, with the SL1 hairpin at the 5' end sufficient to protect from NSP1-mediated degradation at low NSP1 concentrations. We show further that the N-terminal domain of NSP1 is necessary and sufficient for efficient cleavage. We suggest that in the ribosome-bound NSP1 protein the catalytic residues of the N-terminal domain are unmasked by the remodelling of the α1- and α2-helices of the C-terminal domain.
Topics: Humans; COVID-19; Protein Biosynthesis; Ribosomes; RNA, Messenger; SARS-CoV-2; Viral Nonstructural Proteins
PubMed: 37503833
DOI: 10.1093/nar/gkad627 -
Journal of the American College of... Aug 2023To guide implementation of supplemental breast screening by assessing patient preferences for contrast-enhanced mammography (CEM) versus MRI using analytic hierarchy...
OBJECTIVE
To guide implementation of supplemental breast screening by assessing patient preferences for contrast-enhanced mammography (CEM) versus MRI using analytic hierarchy process (AHP) methodology.
METHODS
In an institutional review board-approved, HIPAA-compliant protocol, from March 23 to June 3, 2022, we contacted 579 women who had both CEM screening and MRI. Women were e-mailed an invitation to complete an online survey developed using an AHP-based model to elicit preferences for CEM or MRI. Methods for categorical data analysis were used to evaluate factors affecting preferences, under the Bonferroni correction for multiplicity.
RESULTS
Complete responses were received from 222 (38.3%) women; the 189 women with a personal history of breast cancer had a mean age 61.8 years, and the 34 women without a personal history of breast cancer had a mean age of 53.6 years. Of 222 respondents, 157 (70.7%, confidence interval [CI]: 64.7-76.7) were determined to prefer CEM to MRI. Breast positioning was the most important criterion for 74 of 222 (33.3%) respondents, with claustrophobia, intravenous line placement, and overall stress most important for 38, 37, and 39 women (17.1%, 16.7%, and 17.6%), respectively, and noise level, contrast injection, and indifference being emphasized least frequently (by 10 [4.5%], 11 [5.0%], and 13 [5.9%] women, respectively). CEM preference was most prevalent (MRI least prevalent) for respondents emphasizing claustrophobia (37 of 38 [97%], CI: 86.2-99.9); CEM preference was least prevalent (MRI most prevalent) for respondents emphasizing breast positioning (40 of 74 [54%], CI: 42.1-65.7).
CONCLUSIONS
AHP-based modeling reveals strong patient preferences for CEM over MRI, with claustrophobia favoring preference for CEM and breast positioning relatively favoring preference for MRI. Our results should help guide implementation of screening CEM and MRI.
Topics: Female; Humans; Middle Aged; Breast Neoplasms; Patient Preference; Analytic Hierarchy Process; Early Detection of Cancer; Mammography; Breast; Magnetic Resonance Imaging; Contrast Media
PubMed: 37394083
DOI: 10.1016/j.jacr.2023.05.014 -
Biotechnology and Bioengineering Oct 2023Eukaryotic cell-free protein synthesis (CFPS) can accelerate expression and high-throughput analysis of complex proteins with functionally relevant post-translational...
Eukaryotic cell-free protein synthesis (CFPS) can accelerate expression and high-throughput analysis of complex proteins with functionally relevant post-translational modifications (PTMs). However, low yields and difficulties scaling such systems have prevented their widespread adoption in protein research and manufacturing. Here, we provide detailed demonstrations for the capabilities of a CFPS system derived from Nicotiana tabacum BY-2 cell culture (BY-2 lysate; BYL). BYL is able to express diverse, functional proteins at high yields in 48 h, complete with native disulfide bonds and N-glycosylation. An optimized version of the technology is commercialized as ALiCE® and advances in scaling of BYL production methodologies now allow scaling of eukaryotic CFPS reactions. We show linear, lossless scale-up of batch mode protein expression from 100 µL microtiter plates to 10 and 100 mL volumes in Erlenmeyer flasks, culminating in preliminary data from a litre-scale reaction in a rocking-type bioreactor. Together, scaling across a 20,000x range is achieved without impacting product yields. Production of multimeric virus-like particles from the BYL cytosolic fraction were then shown, followed by functional expression of multiple classes of complex, difficult-to-express proteins using the native microsomes of the BYL CFPS. Specifically: a dimeric enzyme; a monoclonal antibody; the SARS-CoV-2 receptor-binding domain; a human growth factor; and a G protein-coupled receptor membrane protein. Functional binding and activity are demonstrated, together with in-depth PTM characterization of purified proteins through disulfide bond and N-glycan analysis. Taken together, BYL is a promising end-to-end R&D to manufacturing platform with the potential to significantly reduce the time-to-market for high value proteins and biologics.
Topics: Humans; Biotechnology; Nicotiana; COVID-19; SARS-CoV-2; Protein Biosynthesis; Antibodies, Monoclonal; Disulfides; Cell-Free System
PubMed: 37376851
DOI: 10.1002/bit.28461 -
RoFo : Fortschritte Auf Dem Gebiete Der... Nov 2023Sustainability is becoming increasingly important in radiology. Besides climate protection - economic, ecological, and social aspects are integral elements of...
BACKGROUND
Sustainability is becoming increasingly important in radiology. Besides climate protection - economic, ecological, and social aspects are integral elements of sustainability. An overview of the scientific background of the sustainability and environmental impact of radiology as well as possibilities for future concepts for more sustainable diagnostic and interventional radiology are presented below.The three elements of sustainability:1. EcologyWith an annually increasing number of tomographic images, Germany is in one of the leading positions worldwide in a per capita comparison. The energy consumption of an MRI system is comparable to 26 four-person households annually. CT and MRI together make a significant contribution to the overall energy consumption of a hospital. In particular, the energy consumption in the idle or inactive state is responsible for a relevant proportion.2. EconomyA critical assessment of the indications for radiological imaging is important not only because of radiation protection, but also in terms of sustainability and "value-based radiology". As part of the "Choosing Wisely" initiative, a total of 600 recommendations for avoiding unnecessary examinations were compiled from various medical societies, including specific indications in radiological diagnostics.3. Social SustainabilityThe alignment of radiology to the needs of patients and referring physicians is a core aspect of the social component of sustainability. Likewise, ensuring employee loyalty by supporting and maintaining motivation, well-being, and job satisfaction is an essential aspect of social sustainability. In addition, sustainable concepts are of relevance in teaching and research, such as the educational curriculum for residents in radiology, RADUCATION or the recommendations of the International Committee of Medical Journal Editors.
KEY POINTS
· Sustainability comprises three pillars: economy, ecology and the social component.. · Radiologies have a high optimization potential due to a significant demand of these resources.. · A dialogue between medicine, politics and industry is necessary for a sustainable radiology.. · The discourse, knowledge transfer and public communication of recommendations are part of the sustainability network of the German Roentgen Society (DRG)..
CITATION FORMAT
· Palm V, Heye T, Molwitz I et al. Sustainability and Climate Protection in Radiology - An Overview. Fortschr Röntgenstr 2023; 195: 981 - 988.
Topics: Humans; Radiography; Radiology, Interventional; Curriculum; Magnetic Resonance Imaging; Job Satisfaction
PubMed: 37348529
DOI: 10.1055/a-2093-4177 -
RoFo : Fortschritte Auf Dem Gebiete Der... Aug 2023Breast imaging represents an integral part of radiology and is subject to strict quality controls. Regarding this, precise diagnostics including multimodal assessment by...
PURPOSE
Breast imaging represents an integral part of radiology and is subject to strict quality controls. Regarding this, precise diagnostics including multimodal assessment by mammography, sonography, and MRI, including image-guided biopsy and localization procedures, is often decisive and must be performed by experts with profound knowledge and skills in all of these procedures.However, due to numerous restructurings, breast imaging has been shifted more and more towards large, specialized centers, resulting in less patient exposition and training opportunities for radiologists in smaller sites. The following whitepaper summarizes the current circumstances and discusses opinions of the participating societies.
MATERIALS
Under the leadership of the German Roentgen Society (DRG) and with the participation of the DRG's AG Mammadiagnostik, the CAFRAD (Chefarztforum Radiologie), the KLR (Konferenz der Lehrstuhlinhaber für Radiologie e. V.), the DRG's Forum Junge Radiologie (FJR) and the Berufsverband der Deutschen Radiologen e. V. (BDR), possible solutions were discussed and consented for a structured training in breast radiology in the future.
RESULTS
In addition to the teaching provided at the primary workplace, qualified training should be ensured through flexible, multi-institutional, interdisciplinary, and cross-sectoral collaboration. Furthermore, the integration of online case collections and close cooperation with certified breast cancer centers and mammography screening units is recommended. It is indispensible that online courses and case collections adhere to the standards of the national societies and include a maximum of one third of the required cases.
CONCLUSION
In order to provide training in breast radiology at a high professional level, a paradigm shift with closer cooperation of all participants is necessary. This includes close collaboration of the breast imaging societies with the federal medical associations to establish new teaching concepts like e-learning in the training schedule of radiologists.
KEY POINTS
· Breast diagnostics is an integral part of radiology training.. · Due to recent restructurings, smaller training centers have difficulties in meeting the case numbers demanded by the Specialist Training Regulations (WBO). Improved integration of the new structures and their adaptation to the needs of education are necessary to guarantee standardized high-quality training of young radiologists.. · The integration of certified case collections enables quality-assured training, even across regions in online-based formats. In accordance with the "blended learning principle", up to one-third of the required number of patient studies can be substituted with cases from a certified case collection.. · Legally secured short- and medium-term internships may complement training in radiology..
CITATION FORMAT
· Sauer ST, Bley TA, Wenkel E et al. Whitepaper: Training in Diagnostic and Interventional Breast Radiology. Fortschr Röntgenstr 2023; 195: 699 - 706.
Topics: Humans; Radiology, Interventional; Curriculum; Learning; Mammography; Ultrasonography
PubMed: 37348527
DOI: 10.1055/a-2081-3549 -
Australian Endodontic Journal : the... Aug 2023This study aimed to develop a predictive model to screen for undetected vertical root fractures (VRFs) in root canal treated teeth. We included 95 root canal treated...
This study aimed to develop a predictive model to screen for undetected vertical root fractures (VRFs) in root canal treated teeth. We included 95 root canal treated teeth with suspected VRFs; 77 for training and 18 for validation. Following clinical and cone-beam CT parameters were recorded: sex, tooth type, coronal restoration, time interval from completion of endodontic treatment to definitive diagnosis (TI), type of bone loss (BL), apical extent of root filling (AR) and the ratio of root filling diameter to the actual diameter in the coronal (1/3TA) and middle (2/3TA) root thirds. A predictive model p = 1/(1 - e ) was generated, where x = -7.433 + 1.977BL + 1.479 (2/3TA) + 1.102 AR; the sensitivity and specificity were 0.852 and 0.875 for training and 0.917 and 0.833 for validation. VRF teeth were more likely to have vertical bone loss and overfilled root canals. This model had a high diagnostic efficacy for VRFs.
Topics: Humans; Tooth Root; Tooth Fractures; Root Canal Therapy; Fractures, Bone; Sensitivity and Specificity; Tooth, Nonvital; Cone-Beam Computed Tomography; Bone Diseases, Metabolic
PubMed: 35861533
DOI: 10.1111/aej.12667