33 research outputs found
Parallel Optimisation of Potency and Pharmacokinetics Leading to the Discovery of a Pyrrole Carboxamide ERK5 Chemical Tool
[Image: see text] The nonclassical extracellular signal-related kinase 5 (ERK5) mitogen-activated protein kinase pathway has been implicated in increased cellular proliferation, migration, survival, and angiogenesis; hence, ERK5 inhibition may be an attractive approach for cancer treatment. However, the development of selective ERK5 inhibitors has been challenging. Previously, we described the development of a pyrrole carboxamide high-throughput screening hit into a selective, submicromolar inhibitor of ERK5 kinase activity. Improvement in the ERK5 potency was necessary for the identification of a tool ERK5 inhibitor for target validation studies. Herein, we describe the optimization of this series to identify nanomolar pyrrole carboxamide inhibitors of ERK5 incorporating a basic center, which suffered from poor oral bioavailability. Parallel optimization of potency and in vitro pharmacokinetic parameters led to the identification of a nonbasic pyrazole analogue with an optimal balance of ERK5 inhibition and oral exposure
Clinical-grade Detection of Microsatellite Instability in Colorectal Tumors by Deep Learning
Background and Aims: Microsatellite instability (MSI) and mismatch-repair deficiency (dMMR) in colorectal tumors are used to select treatment for patients. Deep learning can detect MSI and dMMR in tumor samples on routine histology slides faster and cheaper than molecular assays. But clinical application of this technology requires high performance and multisite validation, which have not yet been performed.
Methods: We collected hematoxylin and eosin-stained slides, and findings from molecular analyses for MSI and dMMR, from 8836 colorectal tumors (of all stages) included in the MSIDETECT consortium study, from Germany, the Netherlands, the United Kingdom, and the United States. Specimens with dMMR were identified by immunohistochemistry analyses of tissue microarrays for loss of MLH1, MSH2, MSH6, and/or PMS2. Specimens with MSI were identified by genetic analyses. We trained a deep-learning detector to identify samples with MSI from these slides; performance was assessed by cross-validation (n=6406 specimens) and validated in an external cohort (n=771 specimens). Prespecified endpoints were area under the receiver operating characteristic (AUROC) curve and area under the precision-recall curve (AUPRC).
Results: The deep-learning detector identified specimens with dMMR or MSI with a mean AUROC curve of 0.92 (lower bound 0.91, upper bound 0.93) and an AUPRC of 0.63 (range, 0.59–0.65), or 67% specificity and 95% sensitivity, in the cross-validation development cohort. In the validation cohort, the classifier identified samples with dMMR with an AUROC curve of 0.95 (range, 0.92–0.96) without image-preprocessing and an AUROC curve of 0.96 (range, 0.93–0.98) after color normalization.
Conclusions: We developed a deep-learning system that detects colorectal cancer specimens with dMMR or MSI using hematoxylin and eosin-stained slides; it detected tissues with dMMR with an AUROC of 0.96 in a large, international validation cohort. This system might be used for high-throughput, low-cost evaluation of colorectal tissue specimens
<em>In situ</em> magnetic resonance imaging of electrically-induced water diffusion in a Nafion ionic polymer film
Comparative analysis of isocentric 3-dimensional C-arm fluoroscopy and biplanar fluoroscopy for anterior screw fixation in odontoid fractures
STUDY DESIGN: Retrospective clinical study. OBJECTIVE: To compare long-term radiographic and clinical outcomes of patients undergoing anterior odontoid screw placement using traditional biplanar fluoroscopy or isocentric 3-dimensional C-arm (iso-C) fluoroscopy-assisted techniques. SUMMARY OF BACKGROUND DATA: Anterior screw fixation of odontoid fractures preserves motion at the C1-C2 joint, but accurate screw positioning is essential for successful outcomes. Biplanar fluoroscopy image guidance is most often used; however, iso-C imaging improves the ease and accuracy of screw placement with less radiation exposure. METHODS: Fifty-one patients underwent anterior odontoid screw fixation for type II (48 patients) and rostral type III fractures (3 patients). Procedures were guided by biplanar fluoroscopy in 25 (49%) patients, and with iso-C assistance in 26 (51%). Length of surgery, complications, and clinical outcomes based on the Smiley-Webster score were evaluated. Computed tomography confirmed adequate screw placement. Follow-up ranged from 3 to 9 months. RESULTS: At 3-month follow-up, screw position and fusion across the fracture were evident in 87% of the cases treated with biplanar fluoroscopy and in 100% treated by iso-C. The average outcome score in the iso-C group was superior to that of the biplanar group (1.08 vs. 1.33, respectively), although not statistically significant. At last follow-up, the rate of successful fusion was 88% in the biplanar group and 95% in the iso-C group. Length of surgery was significantly lower in the iso-C group compared with the biplanar group (P=0.05). The significantly longer preparation time in the iso-C group (P=0.04) accounted for no overall difference in total operating room occupancy time between the 2 groups. CONCLUSIONS: Iso-C significantly decreased surgical time. At last follow-up iso-C assistance was associated with improved rates of radiographic fusion with comparable outcome and complication profiles. This series represents the largest cohort of patients treated with intraoperative real-time navigation assistance for odontoid fractures
A history of the Barrow Neurological Institute.
The Barrow Neurological Institute (BNI), founded in 1961, is in partnership with St. Joseph\u27s Hospital and Medical Center and part of the Catholic Healthcare West system. The BNI is a relative newcomer to academic neuroscience. However, since its inception it has grown to become an international destination for neurologic disease. This article describes the history of the institute as it has grown over the years in its commitment to excellence in patient care, education, and research
Detection of the cyanobacterial toxin, microcystin-LR, using a novel recombinant antibody-based optical-planar waveguide platform
Microcystins are a major group of cyanobacterial heptapeptide toxins found in freshwater and brackish environments. There is currently an urgent requirement for highly-sensitive, rapid and in-expensive detection methodologies for these toxins. A novel single chain fragment variable (scFv) fragment was generated and is the first known report of a recombinant anti-microcystin avian antibody. In a surface plasmon resonance-based immunoassay, the antibody fragment displayed cross-reactivity with seven microcystin congeners (microcystin-leucine-arginine (MC-LR) 100%, microcystin-tyrosine-arginine (MC-YR) 79.7%, microcystin-leucine-alanine (MC-LA) 74.8%, microcystin-leucine-phenylalanine (MC-LF) 67.5%, microcystin-leucine-tryptophan (MC-LW) 63.7%, microcystin-arginine-arginine (MC-RR) 60.1% and nodularin (Nod) 69.3%, % cross reactivity). Following directed molecular evolution of the parental clone the resultant affinity-enhanced antibody fragment was applied in an optimized fluorescence immunoassay on a planar waveguide detection system. This novel immuno-sensing format can detect free microcystin-LR with a functional limit of detection of 0.19ngmL-1and a detection range of 0.21-5.9ngmL-1. The assay is highly reproducible (displaying percentage coefficients of variance below 8% for intra-day assays and below 11% for inter-day assays), utilizes an inexpensive cartridge system with low reagent volumes and can be completed in less than twenty minutes.</p
Identification of A2B5+CD133- tumor-initiating cells in adult human gliomas.
OBJECTIVE: Several studies have shown that human gliomas contain a small population of cells with stem cell-like features. It has been proposed that these cancer stem cells may be uniquely responsible for glioma formation and recurrence. However, human gliomas also contain an abundance of cells that closely resemble more differentiated glial progenitors. Animal model studies have shown that these cells also possess the capacity to form malignant gliomas.
METHODS: To investigate the contributions of stem-like and progenitor-like cells in human gliomas, we used flow cytometry to characterize the expression of a cancer stem cell marker (CD133) and a glial progenitor marker (A2B5) in 25 tumors. We found that human gliomas consistently express A2B5 in a large percentage of cells (61.7 +/- 3.8%, standard error of the mean). In contrast, CD133 expression was less abundant and less consistent (14.8 +/- 3.6%, standard error of the mean), with several glioblastomas containing very few or no detectable CD133+ cells. When present, the CD133+ population was almost entirely contained within the A2B5+ population. Thus, most gliomas could be divided into three distinct populations on the basis of these markers (A2B5+CD133+, A2B5+CD133-, and A2B5-CD133-). To test the tumorigenic potential of these populations, we separated cells from six tumors by fluorescence-activated cell sorting and reinjected them into nude rats.
RESULTS: We found that the capacity for these different populations to form tumors varied depending on the human tumor specimen from which they were isolated. Of the six human gliomas tested, four contained A2B5+/CD133- cells that formed tumors when transplanted into nude rats, three contained A2B5+/CD133+ cells that formed tumors, and only one glioma contained A2B5-/CD133- cells with the capacity to form tumors.
CONCLUSION: Together, these results demonstrate that human gliomas contain multiple populations of cells with the capacity to form tumors and specifically identify a population of tumorigenic A2B5+ cells that are phenotypically distinct from CD133+ cells
CRAF Autophosphorylation of Serine 621 Is Required to Prevent Its Proteasome-Mediated Degradation
Sex or sanctuary: How do asexual worms survive the winter?
10.1007/s10750-005-0700-9Hydrobiologia5591395-399HYDR