Zobrazení zdroje stránky Mccormickfrandsen1466

High-throughput cytostatic and cell death assays are a critical component of pharmacological screens and mechanism-based interrogations into cellular biology. We developed a method for single-cell and population-level analyses using real-time kinetic labeling (abbreviated "SPARKL") with non-toxic fluorescent probes and high-content live-cell imagers. The protocols herein detail the steps, specifics, and suggested utilization of the SPARKL method within several "label-and-go" zero-handling workflows. For complete details on the use and execution of this protocol, please refer to Gelles et al. (2019).Ageing, and particularly the onset of age-related diseases, is associated with tissue dysfunction and macromolecular damage, some of which can be attributed to accumulation of oxidative damage. Polyphenolic natural products such as stilbenoids, flavonoids and chalcones have been shown to be effective at ameliorating several age-related phenotypes, including oxidative stress, inflammation, impaired proteostasis and cellular senescence, both in vitro and in vivo. Here we aim to identify the structural basis underlying the pharmacology of polyphenols towards ROS and related biochemical pathways involved in age-related disease. We compile and describe SAR trends across different polyphenol chemotypes including stilbenoids, flavonoids and chalcones, review their different molecular targets and indications, and identify common structural ground between chemotypes and mechanisms of action. In particular, we focus on the structural requirements for the direct scavenging of reactive oxygen/nitrogen species such as radicals as well as coordination of a broader antioxidant response. We further suggest that it is important to consider multiple (rather than single) biological activities when identifying and developing new medicinal chemistry entities with utility in modulating complex biological properties such as cell ageing.Relapse of acute lymphoblastic leukemia (ALL) remains a leading cause of childhood death. Prior studies have shown clonal mutations at relapse often arise from relapse-fated subclones that exist at diagnosis. However, the genomic landscape, evolutionary trajectories and mutational mechanisms driving relapse are incompletely understood. In an analysis of 92 cases of relapsed childhood ALL, incorporating multimodal DNA and RNA sequencing, deep digital mutational tracking and xenografting to formally define clonal structure, we identify 50 significant targets of mutation with distinct patterns of mutational acquisition or enrichment. CREBBP, NOTCH1, and Ras signaling mutations rose from diagnosis subclones, whereas variants in NCOR2, USH2A and NT5C2 were exclusively observed at relapse. Evolutionary modeling and xenografting demonstrated that relapse-fated clones were minor (50%), major (27%) or multiclonal (18%) at diagnosis. Putative second leukemias, including those with lineage shift, were shown to most commonly represent relapse from an ancestral clone rather than a truly independent second primary leukemia. A subset of leukemias prone to repeated relapse exhibited hypermutation driven by at least three distinct mutational processes, resulting in heightened neoepitope burden and potential vulnerability to immunotherapy. Finally, relapse-driving sequence mutations were detected prior to relapse using deep digital PCR at levels comparable to orthogonal approaches to monitor levels of measurable residual disease. These results provide a genomic framework to anticipate and circumvent relapse by earlier detection and targeting of relapse-fated clones.<br /><br /> To evaluate the diagnostic utility of two convolutional neural networks (CNNs) for severity staging of anterior cruciate ligament (ACL) injuries.<br /><br /> In this retrospective study, 1243 knee MR images (1008 intact, 18 partially torn, 77 fully torn, and 140 reconstructed ACLs) from 224 patients (mean age, 47 years ± 14 [standard deviation]; 54% women) were analyzed. The MRI examinations were performed between 2011 and 2014. A modified scoring metric was used. Classification of ACL injuries using deep learning involved use of two types of CNN, one with three-dimensional (3D) and the other with two-dimensional (2D) convolutional kernels. Performance metrics included sensitivity, specificity, weighted Cohen κ, and overall accuracy, and the McNemar test was used to compare the performance of the CNNs.<br /><br /> The overall accuracies for ACL injury classification using the 3D CNN and 2D CNN were 89% (225 of 254) and 92% (233 of 254), respectively (<br /><br /> = .27), and both CNNs had a weighted Cohen κ of 0.83. The 2D CNN and 3D CNN performed similarly in classifying intact ACLs (2D CNN, sensitivity of 93% [188 of 203] and specificity of 90% [46 of 51] vs 3D CNN, sensitivity of 89% [180 of 203] and specificity of 88% [45 of 51]). Classification of full tears by both networks was also comparable (2D CNN, sensitivity of 82% [14 of 17] and specificity of 94% [222 of 237] vs 3D CNN, sensitivity of 76% [13 of 17] and specificity of 100% [236 of 237]). The 2D CNN classified all reconstructed ACLs correctly.<br /><br /> Two-dimensional and 3D CNNs applied to ACL lesion classification had high sensitivity and specificity, suggesting that these networks could be used to help nonexperts grade ACL injuries. <br /><br /> © RSNA, 2020.<br /><br />Two-dimensional and 3D CNNs applied to ACL lesion classification had high sensitivity and specificity, suggesting that these networks could be used to help nonexperts grade ACL injuries. Supplemental material is available for this article. © RSNA, 2020.<br /><br /> Laser interstitial thermal therapy (LITT) is becoming an increasingly popular technique for the treatment of brain lesions. More minimally invasive that open craniotomy for lesion resection, LITT may be more appropriate for lesions that are harder to access through an open approach, deeper lesions, and for patients who may not tolerate open surgery.<br /><br /> A search of the current primary literature on LITT for brain lesions on PubMed was performed. These studies were reviewed and updates on the radiological, pathological, and long-term outcomes after LITT for brain metastases, primary brain tumors, and radiation necrosis as well as common complications are included.<br /><br /> Larger extent of ablation and LITT as frontline treatment were potential predictors of favorable progression-free and overall survival for primary brain tumors. In brain metastases, larger extent of ablation was more significantly associated with survival benefit, whereas tumor size was a possible predictor. The most common complications after LITT are transient and permanent weakness, cerebral edema, hemorrhage, seizures, and hyponatremia.<br /><br /> Although the current literature is limited by small sample sizes and primarily retrospective studies, LITT is a safe and effective treatment for brain lesions in the correct patient population.<br /><br />Although the current literature is limited by small sample sizes and primarily retrospective studies, LITT is a safe and effective treatment for brain lesions in the correct patient population.<br /><br /> We aimed to explore gaps in the care of meningioma patients that could improve quality of care by better understanding symptoms experienced by patients at various stages of treatment, and afterwards.<br /><br /> A novel 19-item self-administered questionnaire was provided for patients with meningiomas to complete by the American Brain Tumor Association (ABTA) over a 3-month period.<br /><br /> A total of 1852 unique respondents were included. Nearly one-third of all respondents felt they received insufficient information about meningiomas at initial diagnosis (<br /><br /> = 607, 32.9%) and 28.8% (<br /><br /> = 530) believed they received insufficient information about treatment options. In fact, 34.5% of respondents received the majority of their information from the internet and nonhealthcare professionals. The most common concerns after initial diagnosis were risks associated with surgery and/or treatment (36.5%) followed by how the tumor would impact daily life (25%) and the risk of tumor recurrence (12.4%). Respondents indicated that a list of resources available for patients with meningiomas (<br /><br /> = 597, 32.3%) would have been most beneficial in regards to their disease experience after their initial diagnosis. Moreover, we found that a substantial proportion of patients continued to report symptoms long after treatment, with fatigue being the most common compared to before treatment (38.2% vs. 57.7%, <br /><br /> <br /><br /> = 128, <br /><br /> &lt; .001).<br /><br /> Patients with meningiomas exhibit symptoms that continue well after treatment with fatigue and cognitive impairments as the most bothersome. Moreover, patients report key communication gaps that can be addressed to improve their disease experience and care.<br /><br />Patients with meningiomas exhibit symptoms that continue well after treatment with fatigue and cognitive impairments as the most bothersome. Moreover, patients report key communication gaps that can be addressed to improve their disease experience and care.<br /><br /> Polyglutamylation is a reversible protein modification that commonly occurs in tumor cells. #link# Methotrexate (MTX) in tumor cells is polyglutamylated and strongly binds to dihydrofolate reductase (DHFR) without competitive inhibition by leucovorin. link2 Therefore, tumor cells with high polyglutamylation levels are supposed to be selectively killed, whereas normal cells with lower polyglutamylation are rescued by leucovorin. This study investigated the combined effects of MTX plus histone deacetylase inhibitors (HDACIs), which upregulate MTX polyglutamylation, in primary central nervous system lymphoma (PCNSL).<br /><br /> We evaluated cell viability after MTX treatment and leucovorin rescue and compared the expression of folylpolyglutamate synthetase (FPGS), γ-glutamyl hydrolase (GGH), and DHFR in 2 human PCNSL-derived cell lines (HKBML and TK) and a human Burkitt lymphoma cell line (TL-1). Combination treatments were created using 4 HDACIs panobinostat, vorinostat, sodium butyrate, and valproic acid. The expression of DHFR was examined as well as ratios of FPGS/GGH expression. The combined effects of MTX plus HDACIs were evaluated using a cell viability assay, mass spectroscopy imaging, and subcutaneous and intracranial xenograft models.<br /><br /> HDACIs upregulated the ratio of FPGS/GGH expression resulting in increased polyglutamylation of MTX, but also downregulated expression of the target molecule of MTX DHFR. The combination of MTX and vorinostat decreased cell viability in vitro (<br /><br /> &lt; .05) and tumor volumes in a subcutaneous model (<br /><br /> &lt; .0001), and prolonged survival in an intracranial model (<br /><br /> &lt; .01), relative to controls.<br /><br /> HDACIs enhanced the therapeutic effect of MTX through increased polyglutamylation of MTX and concomitant downregulation of DHFR expression.<br /><br />HDACIs enhanced the therapeutic effect of MTX through increased polyglutamylation of MTX and concomitant downregulation of DHFR expression.<br /><br /> Despite maximal therapy with surgery, chemotherapy, and radiotherapy, glioblastoma (GBM) patients have a median survival of only 15 months. Almost all patients inevitably experience symptomatic tumor recurrence. A hallmark of this tumor type is the large heterogeneity between patients and within tumors itself which relates to the failure of standardized tumor treatment. In [https://www.selleckchem.com/products/sbi-477.html SBI-477 clinical trial] , tissue samples of paired primary and recurrent GBM tumors were investigated to identify individual factors related to tumor progression.<br /><br /> Paired primary and recurrent GBM tumor tissues from 8 patients were investigated with a multiomics approach using transcriptomics, proteomics, and phosphoproteomics.<br /><br /> In the studied patient cohort, large variations between and within patients are observed for all omics analyses. link3 A few pathways affected at the different omics levels partly overlapped if patients are analyzed at the individual level, such as synaptogenesis (containing the SNARE complex) and cholesterol metabolism. Phosphoproteomics revealed increased STMN1(S38) phosphorylation as part of ERBB4 signaling.