BRAF and CRAF are critical components of the MAPK signaling pathway which is activated in many cancer types. In approximately 1% of melanomas, BRAF or CRAF are activated through structural ...arrangements. We describe here a metastatic melanoma with a GOLGA4-RAF1 fusion and pathogenic variants in CTNNB1 and CDKN2A. Anti-CTLA4/anti-PD1 combination immunotherapy failed to control tumor progression. In the absence of other actionable variants the patient was administered MEK inhibitor therapy on the basis of its potential action against RAF1 fusions. This resulted in a profound and clinically significant response. We demonstrated that GOLGA4-RAF1 expression was associated with ERK activation, elevated expression of the RAS/RAF downstream co-effector ETV5, and a high Ki67 index. These findings provide a rationale for the dramatic response to targeted therapy. This study shows that thorough molecular characterization of treatment-resistant cancers can identify therapeutic targets and personalize management, leading to improved patient outcomes.
Final 5‐year results of Z‐FAST trial Brufsky, Adam M.; Harker, W. Graydon; Beck, J. Thaddeus ...
Cancer,
1 March 2012, Letnik:
118, Številka:
5
Journal Article
Modulating signaling pathways including Wnt and Hippo can induce cardiomyocyte proliferation in vivo. Applying these signaling modulators to human induced pluripotent stem cell-derived cardiomyocytes ...(hiPSC-CMs) in vitro can expand CMs modestly (<5-fold). Here, we demonstrate massive expansion of hiPSC-CMs in vitro (i.e., 100- to 250-fold) by glycogen synthase kinase-3β (GSK-3β) inhibition using CHIR99021 and concurrent removal of cell-cell contact. We show that GSK-3β inhibition suppresses CM maturation, while contact removal prevents CMs from cell cycle exit. Remarkably, contact removal enabled 10 to 25 times greater expansion beyond GSK-3β inhibition alone. Mechanistically, persistent CM proliferation required both LEF/TCF activity and AKT phosphorylation but was independent from yes-associated protein (YAP) signaling. Engineered heart tissues from expanded hiPSC-CMs showed comparable contractility to those from unexpanded hiPSC-CMs, demonstrating uncompromised cellular functionality after expansion. In summary, we uncovered a molecular interplay that enables massive hiPSC-CM expansion for large-scale drug screening and tissue engineering applications.
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•GSK-3β inhibition-mediated hiPSC-cardiomyocyte proliferation is cell density dependent•GSK-3β inhibition with reduced cell-cell contact massively expands hiPSC-cardiomyocytes•LEF/TCF activity inhibits hiPSC-cardiomyocyte maturation without promoting cell cycling•Long-term expansion does not alter cardiomyocyte contractile function
Deriving a large number of hiPSC-cardiomyocytes would be beneficial for large-scale tissue engineering and drug screening applications. Buikema et al. show that GSK-3β inhibition combined with removal of cell-cell contact enables massive expansion of hiPSC-cardiomyocytes with comparable function to non-expanded cells.
Introduction: In 2013, injuries to bicyclists accounted for 925 fatalities and 493,884 nonfatal, emergency department-treated injuries in the United States. Bicyclist deaths increased by 19% from ...2010 to 2013. The greatest risk of death and disability to bicyclists is head injuries. The objective of this study was to provide estimates of prevalence and associated factors of bicycle riding and helmet use among children and adults in the United States. Method: CDC analyzed self-reported data from the 2012 Summer ConsumerStyles survey. Adult respondents (IS + years) were asked about bicycle riding and helmet use in the last 30 days for themselves and their children (5 to 17 years). For bicycle riders, CIX estimated the prevalence of helmet use and conducted multivariable regression analyses to identify factors associated with helmet use. Results: Among adults, 21% rode bicycles within the past 30 days and 29% always wore helmets. Respondents reported that of the 61% of children who rode bicycles within the past 30 days, 42% always wore helmets. Children were more likely to always wear helmets (90%) when their adult respondents always wore helmets than when their adult respondents did not always wear helmets (38%). Children who lived in states with a child bicycle helmet law were more likely to always wear helmets (47%) than those in states without a law (39%). Conclusions: Despite the fact that bicycle helmets are highly effective at reducing the risk for head injuries, including severe brain injuries and death, less than half of children and adults always wore bicycle helmets while riding. Practical application: States and communities should consider interventions that improve the safety of riding such as policies to promote helmet use, modeling of helmet wearing by adults, and focusing on high risk groups, including Hispanic cyclists, occasional riders, adults, and children ages 10 to 14.
As a new method for the synthesis of chalcogen polycationic clusters, the electrochemical dissolution of elemental tellurium in ionic liquids (IL) or in liquid SO2 is presented. ILs used are ...ethylmethylimidazolium triflate OTf− and tetraalkylammonium triflylimide NTf2−. Tristriflylmethanide CTf3− was used as BuMeImCTf3 as the electrolyte in SO2. This allowed for the isolation of Te4CTf32, Te6OTf4, and Te8NTf22 containing the square Te42+, the prismatic Te64+, and the novel barrelane‐shaped Te82+. The compounds are novel compositions as they do not contain the usual halometalate anions, but rather common weakly coordinating anions. The 125Te NMR spectrum of an IL solution containing Te82+ features only one broad signal at 2700 ppm. DFT calculations show that slight concerted displacements within the Te82+ cluster lead to a fluxional molecular structure and a fast valence isomerism with a very low activation barrier of about 8 kJ mol−1.
Anodic oxidation of elemental tellurium in ionic liquids provides access to the polycationic clusters Te42+, Te64+, and the new cluster Te82+ with a barrelane structure (see picture). The characterization of the Te82+ ion by 125Te NMR spectroscopy in solution showed a dynamic molecule with fast valence isomerism.
Tubular basement membrane (TBM) deposits are very uncommon in non-lupus membranous nephropathy. We report 5 patients with membranous nephropathy and extensive TBM deposits following allogeneic ...hematopoietic cell transplant. Patients presented with nephrotic syndrome (3 also had acute kidney injury) late post-transplant in association with chronic graft-versus-host disease (cGVHD). Kidney biopsies revealed global subepithelial and extensive TBM immune complex deposits, accompanied by acute tubular injury (n = 4) and tubulointerstitial inflammation (n = 4). Proteomic analysis of glomeruli in 4 cases identified PLA2R in 1, with no significant protein spectra for PLA2R, THSD7A, EX1/2, NELL-1, PCDH7, NCAM1, or SEMA3B detected in the remaining 3. On follow-up (for a mean 42 months), 4 patients had complete and 1 partial remission following prednisone and/or rituximab therapy. We propose that membranous nephropathy with extensive TBM deposits is a distinctive clinicopathologic lesion associated with allogeneic hematopoietic cell transplant. Pathogenesis likely involves cGVHD-driven antibodies against glomerular and TBM components, the identity of which remains to be elucidated.
Despite extensive quantities of work with regard to reactivation of organophosphorus‐inhibited and aged acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), a new therapeutic solution has ...yet to be reached and current methods continue to rely on old oximes and management of the cholinergic crisis as the standard of care. For more information on the latest developments in this highly important field see the Review article by C. M. Hadad et al. on page 5337 ff.
Understanding the biological factors that affect immunological responses may be beneficial in developing more effective bovine respiratory disease vaccination strategies. Dam traits, such as age and ...vaccination history, are not considered when developing vaccination protocols or assessing vaccination efficacy, even though both factors may influence the transference of passive immunity; therefore, a preliminary study was conducted to determine the effect of dam age on the immune response of their calves to vaccinations. Calves born to dams of various ages were vaccinated with a modified live vaccine at 3 to 4 months old, revaccinated with either the same modified live or an inactivated vaccine at weaning, and boosted with the same vaccine 28 days later. Immune and cortisol measures were taken upon initial vaccination, revaccination at weaning, and then 14 and 28 days post-revaccination and post-booster. Overall, calves from older-aged dams began to exhibit a pattern indicative of a dampened immunoglobulin-G response to revaccination and booster, regardless of vaccination type. In contrast, calves from the youngest and oldest dams who received one modified live and two inactivated vaccines had the highest serum-neutralizing titers for most antigens present in the vaccines. These preliminary data partly imply that it may be plausible that the more doses of modified live vaccines the dam receives, which increase with each pregnancy, may affect the immune response of her progeny to vaccination at weaning and revaccination. This may be due to the effects on maternal passive immunity and is further affected by the type of vaccination her progeny received for some measures. However, more research is necessary to elucidate further the effects biological factors have on the immune response to vaccination, develop better vaccination strategies, and further validate these findings through increased sample sizes.
A growing subset of β-secretase (BACE1) inhibitors for the treatment of Alzheimer’s disease (AD) utilizes an anilide chemotype that engages a key residue (Gly230) in the BACE1 binding site. Although ...the anilide moiety affords excellent potency, it simultaneously introduces a third hydrogen bond donor that limits brain availability and provides a potential metabolic site leading to the formation of an aniline, a structural motif of prospective safety concern. We report herein an alternative aminomethyl linker that delivers similar potency and improved brain penetration relative to the amide moiety. Optimization of this series identified analogues with an excellent balance of ADME properties and potency; however, potential drug–drug interactions (DDI) were predicted based on CYP 2D6 affinities. Generation and analysis of key BACE1 and CYP 2D6 crystal structures identified strategies to obviate the DDI liability, leading to compound 16, which exhibits robust in vivo efficacy as a BACE1 inhibitor.
Antimicrobial peptides (AMPs) have therapeutic potential, particularly for localized infections such as those of the lung. Here we show that airway administration of a pegylated AMP minimizes lung ...tissue toxicity while nevertheless maintaining antimicrobial activity. CaLL, a potent synthetic AMP (KWKLFKKIFKRIVQRIKDFLR) comprising fragments of LL-37 and cecropin A peptides, was N-terminally pegylated (PEG-CaLL). PEG-CaLL derivatives retained significant antimicrobial activity (50% inhibitory concentrations IC50s 2- to 3-fold higher than those of CaLL) against bacterial lung pathogens even in the presence of lung lining fluid. Circular dichroism and fluorescence spectroscopy confirmed that conformational changes associated with the binding of CaLL to model microbial membranes were not disrupted by pegylation. Pegylation of CaLL reduced AMP-elicited cell toxicity as measured using in vitro lung epithelial primary cell cultures. Further, in a fully intact ex vivo isolated perfused rat lung (IPRL) model, airway-administered PEG-CaLL did not result in disruption of the pulmonary epithelial barrier, whereas CaLL caused an immediate loss of membrane integrity leading to pulmonary edema. All AMPs (CaLL, PEG-CaLL, LL-37, cecropin A) delivered to the lung by airway administration showed limited (<3%) pulmonary absorption in the IPRL with extensive AMP accumulation in lung tissue itself, a characteristic anticipated to be beneficial for the treatment of pulmonary infections. We conclude that pegylation may present a means of improving the lung biocompatibility of AMPs designed for the treatment of pulmonary infections.
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