The power of forward genetics in yeast is the foundation on which the field of autophagy research firmly stands. Complementary work on autophagy in higher eukaryotes has revealed both the deep ...conservation of this process, as well as novel mechanisms by which autophagy is regulated in the context of development, immunity, and neuronal homeostasis. The recent emergence of new clustered regularly interspaced palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9)-based technologies has begun facilitating efforts to define novel autophagy factors and pathways by forward genetic screening in mammalian cells. Here, we set out to develop an expanded toolkit of autophagy reporters amenable to CRISPR/Cas9 screening. Genome-wide screening of our reporters in mammalian cells recovered virtually all known autophagy-related (ATG) factors as well as previously uncharacterized factors, including vacuolar protein sorting 37 homolog A (VPS37A), transmembrane protein 251 (TMEM251), amyotrophic lateral sclerosis 2 (ALS2), and TMEM41B. To validate this data set, we used quantitative microscopy and biochemical analyses to show that 1 novel hit, TMEM41B, is required for phagophore maturation. TMEM41B is an integral endoplasmic reticulum (ER) membrane protein distantly related to the established autophagy factor vacuole membrane protein 1 (VMP1), and our data show that these two factors play related, albeit not fully overlapping, roles in autophagosome biogenesis. In sum, our work uncovers new ATG factors, reveals a malleable network of autophagy receptor genetic interactions, and provides a valuable resource (http://crispr.deniclab.com) for further mining of novel autophagy mechanisms.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Autophagosome formation requires multiple autophagy‐related (ATG) factors. However, we find that a subset of autophagy substrates remains robustly targeted to the lysosome in the absence of several ...core ATGs, including the LC3 lipidation machinery. To address this unexpected result, we performed genome‐wide CRISPR screens identifying genes required for NBR1 flux in ATG7KO cells. We find that ATG7‐independent autophagy still requires canonical ATG factors including FIP200. However, in the absence of LC3 lipidation, additional factors are required including TAX1BP1 and TBK1. TAX1BP1's ability to cluster FIP200 around NBR1 cargo and induce local autophagosome formation enforces cargo specificity and replaces the requirement for lipidated LC3. In support of this model, we define a ubiquitin‐independent mode of TAX1BP1 recruitment to NBR1 puncta, highlighting that TAX1BP1 recruitment and clustering, rather than ubiquitin binding per se, is critical for function. Collectively, our data provide a mechanistic basis for reports of selective autophagy in cells lacking the lipidation machinery, wherein receptor‐mediated clustering of upstream autophagy factors drives continued autophagosome formation.
SYNOPSIS
Autophagosome formation requires multiple autophagy‐related (ATG) factors. Receptor‐mediated clustering of the autophagy protein RB1CC1/FIP200 around its cargo to promote autophagosome formation provides a mechanistic basis for reports of selective autophagy in cells lacking the LC3 lipidation machinery.
Autophagy receptor TAX1BP1 is able to drive local autophagosome formation in cells that lack core ATGs and LC3 lipidation.
TAX1BP1's ability to cluster FIP200 around NBR1 puncta promotes continued autophagosome formation and enforces selective cargo incorporation.
Both the SKICH domain and a newly identified N domain are required for LC3‐ and UBZ‐independent function of TAX1BP1.
TAX1BP1‐dependent clustering of the autophagy protein RB1CC1/FIP200 around its cargo promotes autophagosome formation and enforces selective cargo incorporation in the absence of core ATG factors and LC3 lipidation.
Previous analysis of this Intergroup trial demonstrated that with a median follow-up among surviving patients of 45.9 months, the concurrent postoperative administration of cisplatin and radiation ...therapy improved local-regional control and disease-free survival of patients who had high-risk resectable head-and-neck carcinomas. With a minimum of 10 years of follow-up potentially now available for all patients, these results are updated here to examine long-term outcomes.
A total of 410 analyzable patients who had high-risk resected head-and-neck cancers were prospectively randomized to receive either radiation therapy (RT: 60 Gy in 6 weeks) or identical RT plus cisplatin, 100 mg/m(2)i.v. on days 1, 22, and 43 (RT + CT).
At 10 years, the local-regional failure rates were 28.8% vs 22.3% (P=.10), disease-free survival was 19.1% vs 20.1% (P=.25), and overall survival was 27.0% vs 29.1% (P=.31) for patients treated by RT vs RT + CT, respectively. In the unplanned subset analysis limited to patients who had microscopically involved resection margins and/or extracapsular spread of disease, local-regional failure occurred in 33.1% vs 21.0% (P=.02), disease-free survival was 12.3% vs 18.4% (P=.05), and overall survival was 19.6% vs 27.1% (P=.07), respectively.
At a median follow-up of 9.4 years for surviving patients, no significant differences in outcome were observed in the analysis of all randomized eligible patients. However, analysis of the subgroup of patients who had either microscopically involved resection margins and/or extracapsular spread of disease showed improved local-regional control and disease-free survival with concurrent administration of chemotherapy. The remaining subgroup of patients who were enrolled only because they had tumor in 2 or more lymph nodes did not benefit from the addition of CT to RT.
Radiotherapy with concomitant and adjuvant temozolomide is the standard of care for newly diagnosed glioblastoma (GBM). O(6)-methylguanine-DNA methyltransferase (MGMT) methylation status may be an ...important determinant of treatment response. Dose-dense (DD) temozolomide results in prolonged depletion of MGMT in blood mononuclear cells and possibly in tumor. This trial tested whether DD temozolomide improves overall survival (OS) or progression-free survival (PFS) in patients with newly diagnosed GBM.
This phase III trial enrolled patients older than age 18 years with a Karnofsky performance score of ≥ 60 with adequate tissue. Stratification included clinical factors and tumor MGMT methylation status. Patients were randomly assigned to standard temozolomide (arm 1) or DD temozolomide (arm 2) for 6 to 12 cycles. The primary end point was OS. Secondary analyses evaluated the impact of MGMT status.
A total of 833 patients were randomly assigned to either arm 1 or arm 2 (1,173 registered). No statistically significant difference was observed between arms for median OS (16.6 v 14.9 months, respectively; hazard ratio HR, 1.03; P = .63) or median PFS (5.5 v 6.7 months; HR, 0.87; P = .06). Efficacy did not differ by methylation status. MGMT methylation was associated with improved OS (21.2 v 14 months; HR, 1.74; P < .001), PFS (8.7 v 5.7 months; HR, 1.63; P < .001), and response (P = .012). There was increased grade ≥ 3 toxicity in arm 2 (34% v 53%; P < .001), mostly lymphopenia and fatigue.
This study did not demonstrate improved efficacy for DD temozolomide for newly diagnosed GBM, regardless of methylation status. However, it did confirm the prognostic significance of MGMT methylation. Feasibility of large-scale accrual, prospective tumor collection, and molecular stratification was demonstrated.
Understanding potential trajectories in health and drivers of health is crucial to guiding long-term investments and policy implementation. Past work on forecasting has provided an incomplete ...landscape of future health scenarios, highlighting a need for a more robust modelling platform from which policy options and potential health trajectories can be assessed. This study provides a novel approach to modelling life expectancy, all-cause mortality and cause of death forecasts —and alternative future scenarios—for 250 causes of death from 2016 to 2040 in 195 countries and territories.
We modelled 250 causes and cause groups organised by the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) hierarchical cause structure, using GBD 2016 estimates from 1990–2016, to generate predictions for 2017–40. Our modelling framework used data from the GBD 2016 study to systematically account for the relationships between risk factors and health outcomes for 79 independent drivers of health. We developed a three-component model of cause-specific mortality: a component due to changes in risk factors and select interventions; the underlying mortality rate for each cause that is a function of income per capita, educational attainment, and total fertility rate under 25 years and time; and an autoregressive integrated moving average model for unexplained changes correlated with time. We assessed the performance by fitting models with data from 1990–2006 and using these to forecast for 2007–16. Our final model used for generating forecasts and alternative scenarios was fitted to data from 1990–2016. We used this model for 195 countries and territories to generate a reference scenario or forecast through 2040 for each measure by location. Additionally, we generated better health and worse health scenarios based on the 85th and 15th percentiles, respectively, of annualised rates of change across location-years for all the GBD risk factors, income per person, educational attainment, select intervention coverage, and total fertility rate under 25 years in the past. We used the model to generate all-cause age-sex specific mortality, life expectancy, and years of life lost (YLLs) for 250 causes. Scenarios for fertility were also generated and used in a cohort component model to generate population scenarios. For each reference forecast, better health, and worse health scenarios, we generated estimates of mortality and YLLs attributable to each risk factor in the future.
Globally, most independent drivers of health were forecast to improve by 2040, but 36 were forecast to worsen. As shown by the better health scenarios, greater progress might be possible, yet for some drivers such as high body-mass index (BMI), their toll will rise in the absence of intervention. We forecasted global life expectancy to increase by 4·4 years (95% UI 2·2 to 6·4) for men and 4·4 years (2·1 to 6·4) for women by 2040, but based on better and worse health scenarios, trajectories could range from a gain of 7·8 years (5·9 to 9·8) to a non-significant loss of 0·4 years (–2·8 to 2·2) for men, and an increase of 7·2 years (5·3 to 9·1) to essentially no change (0·1 years –2·7 to 2·5) for women. In 2040, Japan, Singapore, Spain, and Switzerland had a forecasted life expectancy exceeding 85 years for both sexes, and 59 countries including China were projected to surpass a life expectancy of 80 years by 2040. At the same time, Central African Republic, Lesotho, Somalia, and Zimbabwe had projected life expectancies below 65 years in 2040, indicating global disparities in survival are likely to persist if current trends hold. Forecasted YLLs showed a rising toll from several non-communicable diseases (NCDs), partly driven by population growth and ageing. Differences between the reference forecast and alternative scenarios were most striking for HIV/AIDS, for which a potential increase of 120·2% (95% UI 67·2–190·3) in YLLs (nearly 118 million) was projected globally from 2016–40 under the worse health scenario. Compared with 2016, NCDs were forecast to account for a greater proportion of YLLs in all GBD regions by 2040 (67·3% of YLLs 95% UI 61·9–72·3 globally); nonetheless, in many lower-income countries, communicable, maternal, neonatal, and nutritional (CMNN) diseases still accounted for a large share of YLLs in 2040 (eg, 53·5% of YLLs 95% UI 48·3–58·5 in Sub-Saharan Africa). There were large gaps for many health risks between the reference forecast and better health scenario for attributable YLLs. In most countries, metabolic risks amenable to health care (eg, high blood pressure and high plasma fasting glucose) and risks best targeted by population-level or intersectoral interventions (eg, tobacco, high BMI, and ambient particulate matter pollution) had some of the largest differences between reference and better health scenarios. The main exception was sub-Saharan Africa, where many risks associated with poverty and lower levels of development (eg, unsafe water and sanitation, household air pollution, and child malnutrition) were projected to still account for substantive disparities between reference and better health scenarios in 2040.
With the present study, we provide a robust, flexible forecasting platform from which reference forecasts and alternative health scenarios can be explored in relation to a wide range of independent drivers of health. Our reference forecast points to overall improvements through 2040 in most countries, yet the range found across better and worse health scenarios renders a precarious vision of the future—a world with accelerating progress from technical innovation but with the potential for worsening health outcomes in the absence of deliberate policy action. For some causes of YLLs, large differences between the reference forecast and alternative scenarios reflect the opportunity to accelerate gains if countries move their trajectories toward better health scenarios—or alarming challenges if countries fall behind their reference forecasts. Generally, decision makers should plan for the likely continued shift toward NCDs and target resources toward the modifiable risks that drive substantial premature mortality. If such modifiable risks are prioritised today, there is opportunity to reduce avoidable mortality in the future. However, CMNN causes and related risks will remain the predominant health priority among lower-income countries. Based on our 2040 worse health scenario, there is a real risk of HIV mortality rebounding if countries lose momentum against the HIV epidemic, jeopardising decades of progress against the disease. Continued technical innovation and increased health spending, including development assistance for health targeted to the world's poorest people, are likely to remain vital components to charting a future where all populations can live full, healthy lives.
Bill & Melinda Gates Foundation.
Patients with grade 2 glioma were randomly assigned to radiation therapy alone or radiation therapy plus six cycles of chemotherapy. The median overall survival with radiation therapy plus ...chemotherapy was 13.3 years, as compared with 7.8 years with radiation therapy alone.
Grade 2 gliomas are relatively uncommon, constituting 5 to 10% of all primary brain tumors in adults. Progressive neurologic symptoms eventually develop in nearly all patients, and nearly all patients die prematurely. At the time of the initiation of our trial, studies had shown that chemotherapy caused tumor regressions in patients with recurrent low-grade gliomas, with regimens that included procarbazine, lomustine (also called CCNU), and vincristine,
1
carmustine (also called BCNU) plus interferon,
2
and mechlorethamine, vincristine, and procarbazine.
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Similarly, the combination of procarbazine, CCNU, and vincristine, when administered as initial therapy, has been shown to result in tumor regressions.
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The . . .
Abstract
Many studies over the past several decades have attempted to correlate trends in lightning (e.g., rates, polarity) to severe weather occurrence. These studies mainly used cloud-to-ground ...(CG) lightning information due to the ease of data availability, high detection efficiency, and broad coverage across the United States, with somewhat inconclusive results. Conversely, it has been demonstrated that trends in total lightning are more robustly correlated to severe weather occurrence, with rapid increases in total lightning observed 10s of minutes prior to the onset of severe weather. Unfortunately, total lightning observations are not as numerous, or available over the same areal coverage domain, as provided by CG networks. Relatively few studies have examined concurrent trends in both total and CG lightning within the same severe thunderstorm, or even large sets of thunderstorms using an objective lightning jump algorithm. Multiple studies have shown that the total flash rate rapidly increases prior to the onset of severe weather. What is untested within the same framework is the use of CG information to perform the same task. Herein, total and CG lightning trends for 711 thunderstorms occurring in four regions of the country were examined to demonstrate the increased utility that total lightning provides over CG lightning, specifically within the framework of developing a useful lightning-based severe weather warning decision support tool. Results indicate that while both lightning datasets demonstrate the presence of increased lightning activity prior to the onset of severe weather, the use of total lightning trends was more effective than CG trends probability of detection (POD), 79% versus 66%; false alarm rate (FAR), 36% versus 53%; critical success index (CSI), 55% versus 38%; Heidke skill score (HSS), 0.71 versus 0.55. Moreover, 40% of false alarms associated with total lightning, and 16% of false alarms with CG lightning trends, occurred when a lightning jump associated with a severe weather “warning” was already in effect. If these false alarms are removed, the FAR drops from 36% to 22% for total lightning and from 53% to 44% for CG lightning. Importantly, average lead times prior to severe weather occurrence were higher using total lightning as compared with CG lightning (20.65 versus 13.54 min). The ultimate goal of this study was to demonstrate the increased utility of total lightning information that the Geostationary Lightning Mapper (GLM) will provide to operational meteorology in anticipation of severe convective weather on a hemispheric scale once Geostationary Operational Environmental Satellite-R (GOES-R) is deployed in the next decade.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
Abstract
A detailed case study analysis of four thunderstorms is performed using polarimetric and multi-Doppler capabilities to provide specificity on the physical and dynamical drivers behind ...lightning jumps. The main differences between small increases in the total flash rate and a lightning jump are the increases in graupel mass and updraft volumes ≥10 m s
−1
between the −10° and −40°C isotherms. Updraft volumes ≥10 m s
−1
increased in magnitude at least 3–5 min in advance of the increase in both graupel mass and total flash rate. Updraft volumes ≥10 m s
−1
are more robustly correlated to total flash rate than maximum updraft speed over a thunderstorm’s entire life cycle. However, peak updraft speeds increase prior to 8 of the 12 lightning jumps examined. Decreases in mean and median flash footprint size during increases in total lightning are observed in all four thunderstorms and are most notable during development stages within the most intense storms. However, this inverse relationship breaks down on larger storm scales as storms mature and anvils and stratiform regions developed with time. Promisingly, smaller flash sizes are still collocated with the strongest updraft speeds, while larger flash sizes are observed within weaker updraft regions. The results herein emphasize the following for lightning jump applications: both the lightning jump sigma level and the resultant magnitude of the total flash rate must be employed in conjunction to assess storm intensity using lightning data. The sigma-level magnitude of the lightning jump is the early warning that indicates that rapid intensification is occurring, while the magnitude of the total flash rate provides insight into the size and maintenance of the updraft volume and graupel mass. These cases serve as conceptual models for future applications of the lightning jump algorithm for hazardous weather monitoring.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
Objective: Cannabis is the most used psychoactive substance among adolescents and is a public health concern. Cannabis demand is a quantifiable measure of the reinforcing value of cannabis and ...comprises two latent-factors-amplitude (maximum consumption) and persistence (sensitivity to increasing costs). Cannabis demand and cannabis motives are important predictors of adolescent cannabis use and associated problems; however, little is known about how these two facets of motivation are causally related. Cannabis motives are thought to represent the final common pathway to cannabis use and may explain why elevated demand is associated with use and consequences. The present study tested whether internal cannabis motives (coping and enjoyment) mediated longitudinal associations between cannabis demand, use (hours high), and negative consequences. Method: Participants aged 15-18 years old (n = 89, Mage = 17.0, SD = 0.9) who reported lifetime cannabis use completed online assessments of cannabis demand, motives, use and negative consequences at baseline, 3-month, and 6-month follow-up. Results: PROCESS mediation models revealed that enjoyment motives mediated the association between amplitude and persistence and use. In addition, coping motives mediated the association between amplitude and negative consequences. Conclusions: These findings suggest that internal motives, while differentially related to aspects of demand and cannabis outcomes, are important in understanding adolescent cannabis use. Prevention efforts aimed at limiting access to cannabis and increasing access to substance-free activities may be important targets for adolescents. Further, cannabis interventions targeting specific motives for using (e.g., to cope with negative affect) may be important for reducing cannabis demand.
Public Health Significance Statement
Findings from the present study suggest that internal cannabis use motives (coping and enjoyment) mediated associations between cannabis demand and cannabis-related outcomes. These findings have important implications for prevention and intervention of adolescent cannabis use. Efforts to increase access to substance-free reinforcers, particularly reinforcers targeting specific cannabis use motives, may be important for reducing cannabis use and associated negative consequences.
Neural mechanisms of observational learning Burke, Christopher J.; Tobler, Philippe N.; Baddeley, Michelle ...
Proceedings of the National Academy of Sciences - PNAS,
08/2010, Letnik:
107, Številka:
32
Journal Article
Recenzirano
Odprti dostop
Individuals can learn by interacting with the environment and experiencing a difference between predicted and obtained outcomes (prediction error). However, many speciesalso learnby observing the ...actions and outcomes of others. In contrast to individual learning, observational learning cannot be based on directly experienced outcome prediction errors. Accordingly, the behavioral and neural mechanisms of learning through observation remain elusive. Here we propose that human observational learning can be explained by two previously uncharacterized forms of prediction error, observational action prediction errors (the actual minus the predicted choice of others) and observational outcome prediction errors (the actual minus predicted outcome received by others). In a functional MRI experiment, we found that brain activity in the dorsolateral prefrontal cortex and the ventromedial prefrontal cortex respectively corresponded to these two distinct observational learning signals.
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