Macrophages activated by the TLR4 agonist LPS undergo dramatic changes in their metabolic activity. We here show that LPS induces expression of the key metabolic regulator Pyruvate Kinase M2 (PKM2). ...Activation of PKM2 using two well-characterized small molecules, DASA-58 and TEPP-46, inhibited LPS-induced Hif-1α and IL-1β, as well as the expression of a range of other Hif-1α-dependent genes. Activation of PKM2 attenuated an LPS-induced proinflammatory M1 macrophage phenotype while promoting traits typical of an M2 macrophage. We show that LPS-induced PKM2 enters into a complex with Hif-1α, which can directly bind to the IL-1β promoter, an event that is inhibited by activation of PKM2. Both compounds inhibited LPS-induced glycolytic reprogramming and succinate production. Finally, activation of PKM2 by TEPP-46 in vivo inhibited LPS and Salmonella typhimurium-induced IL-1β production, while boosting production of IL-10. PKM2 is therefore a critical determinant of macrophage activation by LPS, promoting the inflammatory response.
Display omitted
•Tetramerization of PKM2 reverses the LPS-induced Warburg effect•PKM2 plays a key role in stabilizing Hif-1α and regulates Hif-1α-dependent genes•Tetramerization of PKM2 attenuates LPS-induced M1 macrophage traits•PKM2 is a critical determinant of glycolytic reprogramming in macrophages
TLR4-activated macrophages switch their metabolism from oxidative phosphorylation to glycolysis to rapidly provide for the high energy and biosynthetic demands of infection or injury response. Palsson-McDermott identify PKM2 as a critical modulator of IL-1β production and the Warburg effect in LPS-activated macrophages, highlighting it as a target in cancer and inflammation.
Understanding complex tissues requires single-cell deconstruction of gene regulation with precision and scale. Here, we assess the performance of a massively parallel droplet-based method for mapping ...transposase-accessible chromatin in single cells using sequencing (scATAC-seq). We apply scATAC-seq to obtain chromatin profiles of more than 200,000 single cells in human blood and basal cell carcinoma. In blood, application of scATAC-seq enables marker-free identification of cell type-specific cis- and trans-regulatory elements, mapping of disease-associated enhancer activity and reconstruction of trajectories of cellular differentiation. In basal cell carcinoma, application of scATAC-seq reveals regulatory networks in malignant, stromal and immune cells in the tumor microenvironment. Analysis of scATAC-seq profiles from serial tumor biopsies before and after programmed cell death protein 1 blockade identifies chromatin regulators of therapy-responsive T cell subsets and reveals a shared regulatory program that governs intratumoral CD8
T cell exhaustion and CD4
T follicular helper cell development. We anticipate that scATAC-seq will enable the unbiased discovery of gene regulatory factors across diverse biological systems.
This study examined the effects of speed and leg prostheses on mediolateral (ML) foot placement and its variability in sprinters with and without transtibial amputations. We hypothesized that ML foot ...placement variability would: 1. increase with running speed up to maximum speed and 2. be symmetrical between the legs of non-amputee sprinters but asymmetrically greater for the affected leg of sprinters with a unilateral transtibial amputation. We measured the midline of the body (kinematic data) and center of pressure (kinetic data) in the ML direction while 12 non-amputee sprinters and 7 Paralympic sprinters with transtibial amputations (6 unilateral, 1 bilateral) ran across a range of speeds up to maximum speed on a high-speed force measuring treadmill. We quantified ML foot placement relative to the body's midline and its variability. We interpret our results with respect to a hypothesized relation between ML foot placement variability and lateral balance. We infer that greater ML foot placement variability indicates greater challenges with maintaining lateral balance. In non-amputee sprinters, ML foot placement variability for each leg increased substantially and symmetrically across speed. In sprinters with a unilateral amputation, ML foot placement variability for the affected and unaffected leg also increased substantially, but was asymmetric across speeds. In general, ML foot placement variability for sprinters with a unilateral amputation was within the range observed in non-amputee sprinters. For the sprinter with bilateral amputations, both affected legs exhibited the greatest increase in ML foot placement variability with speed. Overall, we find that maintaining lateral balance becomes increasingly challenging at faster speeds up to maximum speed but was equally challenging for sprinters with and without a unilateral transtibial amputation. Finally, when compared to all other sprinters in our subject pool, maintaining lateral balance appears to be the most challenging for the Paralympic sprinter with bilateral transtibial amputations.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Characterizing the transcriptome of individual cells is fundamental to understanding complex biological systems. We describe a droplet-based system that enables 3' mRNA counting of tens of thousands ...of single cells per sample. Cell encapsulation, of up to 8 samples at a time, takes place in ∼6 min, with ∼50% cell capture efficiency. To demonstrate the system's technical performance, we collected transcriptome data from ∼250k single cells across 29 samples. We validated the sensitivity of the system and its ability to detect rare populations using cell lines and synthetic RNAs. We profiled 68k peripheral blood mononuclear cells to demonstrate the system's ability to characterize large immune populations. Finally, we used sequence variation in the transcriptome data to determine host and donor chimerism at single-cell resolution from bone marrow mononuclear cells isolated from transplant patients.
Metastases are the major cause of death from melanoma, a skin cancer that has the fastest rising incidence of any malignancy in the Western world. Molecular pathways that drive melanoblast migration ...in development are believed to underpin the movement and ultimately the metastasis of melanoma. Here we show that mice lacking P-Rex1, a Rac-specific Rho GTPase guanine nucleotide exchange factor, have a melanoblast migration defect during development evidenced by a white belly. Moreover, these P-Rex1(-/-) mice are resistant to metastasis when crossed to a murine model of melanoma. Mechanistically, this is associated with P-Rex1 driving invasion in a Rac-dependent manner. P-Rex1 is elevated in the majority of human melanoma cell lines and tumour tissue. We conclude that P-Rex1 has an important role in melanoblast migration and cancer progression to metastasis in mice and humans.
Adjuvant chemotherapy for resected early-stage non-small-cell lung cancer (NSCLC) provides a modest survival benefit. Bevacizumab, a monoclonal antibody directed against VEGF, improves outcomes when ...added to platinum-based chemotherapy in advanced-stage non-squamous NSCLC. We aimed to evaluate the addition of bevacizumab to adjuvant chemotherapy in early-stage resected NSCLC.
We did an open-label, randomised, phase 3 trial of adult patients (aged ≥18 years) with an Eastern Cooperative Oncology Group performance status of 0 or 1 and who had completely resected stage IB (≥4 cm) to IIIA (defined by the American Joint Committee on Cancer 6th edition) NSCLC. We enrolled patients from across the US National Clinical Trials Network, including patients from the Eastern Cooperative Oncology Group–American College of Radiology Imaging Network (ECOG-ACRIN) affiliates in Europe and from the Canadian Cancer Trials Group, within 6–12 weeks of surgery. The chemotherapy regimen for each patient was selected before randomisation and administered intravenously; it consisted of four 21-day cycles of cisplatin (75 mg/m2 on day 1 in all regimens) in combination with investigator's choice of vinorelbine (30 mg/m2 on days 1 and 8), docetaxel (75 mg/m2 on day 1), gemcitabine (1200 mg/m2 on days 1 and 8), or pemetrexed (500 mg/m2 on day 1). Patients in the bevacizumab group received bevacizumab 15 mg/kg intravenously every 21 days starting with cycle 1 of chemotherapy and continuing for 1 year. We randomly allocated patients (1:1) to group A (chemotherapy alone) or group B (chemotherapy plus bevacizumab), centrally, using permuted blocks sizes and stratified by chemotherapy regimen, stage of disease, histology, and sex. No one was masked to treatment assignment, except the Data Safety and Monitoring Committee. The primary endpoint was overall survival, analysed by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT00324805.
Between June 1, 2007, and Sept 20, 2013, 1501 patients were enrolled and randomly assigned to the two treatment groups: 749 to group A (chemotherapy alone) and 752 to group B (chemotherapy plus bevacizumab). 383 (26%) of 1458 patients (with complete staging information) had stage IB, 636 (44%) had stage II, and 439 (30%) had stage IIIA disease (stage of disease data were missing for 43 patients). Squamous cell histology was reported for 422 (28%) of 1501 patients. All four cisplatin-based chemotherapy regimens were used: 377 (25%) patients received vinorelbine, 343 (23%) received docetaxel, 283 (19%) received gemcitabine, and 497 (33%) received pemetrexed. At a median follow-up of 50·3 months (IQR 32·9–68·0), the estimated median overall survival in group A has not been reached, and in group B was 85·8 months (95% CI 74·9 to not reached); hazard ratio (group B vs group A) 0·99 (95% CI 0·82–1·19; p=0·90). Grade 3–5 toxicities of note (all attributions) that were reported more frequently in group B (the bevacizumab group) than in group A (chemotherapy alone) were overall worst grade (ie, all grade 3–5 toxicities; 496 67% of 738 in group A vs 610 83% of 735 in group B), hypertension (60 8% vs 219 30%), and neutropenia (241 33% vs 275 37%). The number of deaths on treatment did not differ between the groups (15 deaths in group A vs 19 in group B). Of these deaths, three in group A and ten in group B were considered at least possibly related to treatment.
Addition of bevacizumab to adjuvant chemotherapy did not improve overall survival for patients with surgically resected early-stage NSCLC. Bevacizumab does not have a role in this setting and should not be considered as an adjuvant therapy for patients with resected early-stage NSCLC.
National Cancer Institute of the National Institutes of Health.
The engineered outer domain germline targeting version 8 (eOD-GT8) 60-mer nanoparticle was designed to prime VRC01-class HIV-specific B cells that would need to be matured, through additional ...heterologous immunizations, into B cells that are able to produce broadly neutralizing antibodies. CD4 T cell help will be critical for the development of such high-affinity neutralizing antibody responses. Thus, we assessed the induction and epitope specificities of the vaccine-specific T cells from the IAVI G001 phase 1 clinical trial that tested immunization with eOD-GT8 60-mer adjuvanted with AS01
. Robust polyfunctional CD4 T cells specific for eOD-GT8 and the lumazine synthase (LumSyn) component of eOD-GT8 60-mer were induced after two vaccinations with either the 20- or 100-microgram dose. Antigen-specific CD4 T helper responses to eOD-GT8 and LumSyn were observed in 84 and 93% of vaccine recipients, respectively. CD4 helper T cell epitope "hotspots" preferentially targeted across participants were identified within both the eOD-GT8 and LumSyn proteins. CD4 T cell responses specific to one of these three LumSyn epitope hotspots were observed in 85% of vaccine recipients. Last, we found that induction of vaccine-specific peripheral CD4 T cells correlated with expansion of eOD-GT8-specific memory B cells. Our findings demonstrate strong human CD4 T cell responses to an HIV vaccine candidate priming immunogen and identify immunodominant CD4 T cell epitopes that might improve human immune responses either to heterologous boost immunogens after this prime vaccination or to other human vaccine immunogens.
Muscle design constraints preclude simultaneous specialization of the vertebrate locomotor system for explosive and economical force generation. The resulting performance trade-off between power and ...economy has been attributed primarily to individual differences in muscle fiber type composition. While certainly crucial for performance specialization, fiber type likely interacts with muscle architectural parameters, such as fascicle length, to produce this trade-off. Longer fascicles composed of more serial sarcomeres can achieve faster shortening velocities, allowing for greater power production. Long fascicles likely reduce economy, however, because more energy-consuming contractile units are activated for a given force production. We hypothesized that longer fascicles are associated with both increased power production and locomotor cost. In 11 power-trained and 13 endurance-trained recreational athletes, we measured (1) muscle fascicle length via ultrasound in the gastrocnemius lateralis, gastrocnemius medialis and vastus lateralis, (2) maximal power during cycling and countermovement jumps, and (3) running cost of transport. We estimated muscle fiber type non-invasively based on the pedaling rate at which maximal cycling power occurred. As predicted, longer gastrocnemius muscle fascicles were correlated with greater lower-body power production and cost of transport. Multiple regression analyses revealed that variability in maximal power was explained by fiber type (46% for cycling, 24% for jumping) and average fascicle length (20% for cycling, 13% for jumping), while average fascicle length accounted for 15% of the variation in cost of transport. These results suggest that, at least for certain muscles, fascicle length plays an important role in the power versus economy performance trade-off.
There is little known about sexual offenders hospitalized under forensic commitment statutes such as not guilty by reason of insanity (NGRI). We conducted a chart review to delineate the demographic, ...clinical, and legal characteristics of NGRI sexual offenders (n = 68) committed to the California Department of State Hospitals—Napa, including 41 found NGRI for a sexual offense and 27 found NGRI for a nonsexual offense. The two groups did not differ significantly in their demographics, psychiatric diagnoses, victim characteristics, or recidivism risk as measured by the Static‐99R. Those found NGRI for a sexual offense were older at the time of their first criminal and first violent offense, younger at the time of their committing offense, and had fewer prior total convictions and sexual offense convictions. These findings may indicate that sexual offenders found NGRI for a sexual offense are less antisocial than those found NGRI for a nonsexual offense.
Running-specific prostheses (RSP) emulate the spring-like behaviour of biological limbs during human running, but little research has examined the mechanical means by which amputees achieve top ...speeds. To better understand the biomechanical effects of RSP during sprinting, we measured ground reaction forces (GRF) and stride kinematics of elite unilateral trans-tibial amputee sprinters across a range of speeds including top speed. Unilateral amputees are ideal subjects because each amputee's affected leg (AL) can be compared with their unaffected leg (UL). We found that stance average vertical GRF were approximately 9 per cent less for the AL compared with the UL across a range of speeds including top speed (p < 0.0001). In contrast, leg swing times were not significantly different between legs at any speed (p = 0.32). Additionally, AL and UL leg swing times were similar to those reported for non-amputee sprinters. We infer that RSP impair force generation and thus probably limit top speed. Some elite unilateral trans-tibial amputee sprinters appear to have learned or trained to compensate for AL force impairment by swinging both legs rapidly.
PDF
