Activated K-Ras and H-Ras display different interactions with saturable nonraft sites at the surface of live cells

Ras–membrane interactions play important roles in signaling and oncogenesis. H-Ras and K-Ras have nonidentical membrane anchoring moieties that can direct them to different membrane compartments. Ras–lipid raft interactions were reported, but recent studies suggest that activated K-Ras and H-Ras are not raft resident. However, specific interactions of activated Ras proteins with nonraft sites, which may underlie functional differences and phenotypic variation between different Ras isoforms, are unexplored. Here we used lateral mobility studies by FRAP to investigate the membrane interactions of green fluorescent protein–tagged H- and K-Ras in live cells. All Ras isoforms displayed stable membrane association, moving by lateral diffusion and not by exchange with a cytoplasmic pool. The lateral diffusion rates of constitutively active K- and H-Ras increased with their expression levels in a saturable manner, suggesting dynamic association with saturable sites or domains. These sites are distinct from lipid rafts, as the activated Ras mutants are not raft resident. Moreover, they appear to be different for H- and K-Ras. However, wild-type H-Ras, the only isoform preferentially localized in rafts, displayed cholesterol-sensitive interactions with rafts that were independent of its expression level. Our findings provide a mechanism for selective signaling by different Ras isoforms

Association between ambient temperature, particulate air pollution and emergency room visits for conjunctivitis

BACKGROUND: Numerous studies have confirmed the association of ambient temperature and air pollution with a higher risk of morbidities, yet few have addressed their effect on the ocular system. The purpose of this study was to assess the association between temperature, air pollution, and emergency room visits for conjunctivitis. METHODS: In this case-crossover study, the records of all emergency room visits to Soroka University Medical Center (SUMC) from 2009 to 2014 were reviewed for patients with conjunctivitis. Daily exposure to fine and coarse particulate matter and temperature were determined by a hybrid model involving satellite sensors. Mean relative humidity was obtained from the Ministry of Environmental Protection meteorological monitoring station located in Beer-Sheva. RESULTS: Six hundred one patients were diagnosed with conjunctivitis in the SUMC emergency room. We discovered a positive association between temperature increments and incidence of conjunctivitis. The strongest effect was found during summer and autumn, with an immediate (lag0) incidence increase of 8.1% for each 1 degrees C increase in temperature (OR = 1.088, 95%CI: 1.046-1.132) between 24 and 28 degrees C in the summer and 7.2% for each 1 degrees C increase in temperature (OR = 1.072, 95%CI: 1.036-1.108) between 13 and 23 degrees C in the autumn. There was no statistically significant association between fine and coarse particulate matter and conjunctivitis incidence. CONCLUSION: Temperature increases during summer and autumn are significantly associated with an increased risk of conjunctivitis. Conjunctivitis is not associated with non-anthropogenic air pollution. These findings may help community clinics and hospital emergency rooms better predict conjunctivitis cases and will hopefully lead to improved prevention efforts that will lower the financial burden on both the individual and the public

Predicting fine-scale daily NO2 over Mexico city using an ensemble modeling approach

In recent years, there has been growing interest in developing air pollution prediction models to reduce exposure measurement error in epidemiologic studies. However, efforts for localized, fine-scale prediction models have been predominantly focused in the United States and Europe. Furthermore, the availability of new satellite instruments such as the TROPOsopheric Monitoring Instrument (TROPOMI) provides novel opportunities for modeling efforts. We estimated daily ground-level nitrogen dioxide (NO2) concentrations in the Mexico City Metropolitan Area at 1-km2 grids from 2005 to 2019 using a four-stage approach. In stage 1 (imputation stage), we imputed missing satellite NO2 column measurements from the Ozone Monitoring Instrument (OMI) and TROPOMI using the random forest (RF) approach. In stage 2 (calibration stage), we calibrated the association of column NO2 to ground-level NO2 using ground monitors and meteorological features using RF and extreme gradient boosting (XGBoost) models. In stage 3 (prediction stage), we predicted the stage 2 model over each 1-km2 grid in our study area, then ensembled the results using a generalized additive model (GAM). In stage 4 (residual stage), we used XGBoost to model the local component at the 200-m2 scale. The cross-validated R2 of the RF and XGBoost models in stage 2 were 0.75 and 0.86 respectively, and 0.87 for the ensembled GAM. Cross-validated root-mean-squared error (RMSE) of the GAM was 3.95 μg/m3. Using novel approaches and newly available remote sensing data, our multi-stage model presented high cross-validated fits and reconstructs fine-scale NO2 estimates for further epidemiologic studies in Mexico City

A random forest approach to estimate daily particulate matter, nitrogen dioxide, and ozone at fine spatial resolution in Sweden

Air pollution is one of the leading causes of mortality worldwide. An accurate assessment of its spatial and temporal distribution is mandatory to conduct epidemiological studies able to estimate long-term (e.g., annual) and short-term (e.g., daily) health effects. While spatiotemporal models for particulate matter (PM) have been developed in several countries, estimates of daily nitrogen dioxide (NO 2 ) and ozone (O 3 ) concentrations at high spatial resolution are lacking, and no such models have been developed in Sweden. We collected data on daily air pollutant concentrations from routine monitoring networks over the period 2005-2016 and matched them with satellite data, dispersion models, meteorological parameters, and land-use variables. We developed a machine-learning approach, the random forest (RF), to estimate daily concentrations of PM 10 (PM<10 microns), PM 2.5 (PM<2.5 microns), PM 2.5-10 (PM between 2.5 and 10 microns), NO 2 , and O 3 for each squared kilometer of Sweden over the period 2005-2016. Our models were able to describe between 64% (PM 10 ) and 78% (O 3 ) of air pollutant variability in held-out observations, and between 37% (NO 2 ) and 61% (O 3 ) in held-out monitors, with no major differences across years and seasons and better performance in larger cities such as Stockholm. These estimates will allow to investigate air pollution effects across the whole of Sweden, including suburban and rural areas, previously neglected by epidemiological investigation

Rasosomes originate from the Golgi to dispense Ras signals

Ras proteins undergo an incompletely understood trafficking process in the cell. Rasosomes are protein nanoparticles of 80–100 nm diameter that carry lipidated Ras isoforms (H-Ras and N-Ras) as well as their effectors through the cytoplasm and near the plasma membrane (PM). In this study, we identified the subcellular origin of rasosomes and how they spread Ras proteins through the cell. We found no dependency of rasosome formation on galectins, or on the GDP-/GTP-bound state of Ras. We found that significantly more rasosomes are associated with forms of Ras that are localized to the Golgi, namely N-Ras or the singly palmitoylated H-Ras mutant (C181S). To explore the possibility that rasosome originate from the Golgi, we used photoactivatable (PA)-GFP-H-Ras mutants and showed that rasosomes bud from the Golgi in a two-step mechanism. Newly released rasosomes first move in an energy-dependent directed fashion and then convert to randomly diffusing rasosomes. Dual fluorescence time-lapse imaging revealed the appearance of dually labeled rasosomes, indicating a dynamic exchange of cytoplasmic and PM-associated Ras with rasosome-associated Ras. Finally, higher levels of rasosomes correlate with higher levels of ERK phosphorylation, a key marker of Ras downstream signaling. We suggest that H-Ras and N-Ras proteins exchange with rasosomes that can function as carriers of palmitoylated Ras and its signals

Intercellular Transfer of Oncogenic H-Ras at the Immunological Synapse

Immune cells establish dynamic adhesive cell–cell interactions at a specific contact region, termed the immunological synapse (IS). Intriguing features of the IS are the formation of regions of plasma membrane fusion and the intercellular exchange of membrane fragments between the conjugated cells. It is not known whether upon IS formation, intact intracellular proteins can transfer from target cells to lymphocytes to allow the transmission of signals across cell boundaries. Here we show by both FACS and confocal microscopy that human lymphocytes acquire from the cells they scan the inner-membrane protein H-Ras, a G-protein vital for common lymphocyte functions and a prominent participant in human cancer. The transfer was cell contact-dependent and occurred in the context of cell-conjugate formation. Moreover, the acquisition of oncogenic H-RasG12V by natural killer (NK) and T lymphocytes had important biological functions in the adopting lymphocytes: the transferred H-RasG12V induced ERK phosphorylation, increased interferon-γ and tumor necrosis factor-α secretion, enhanced lymphocyte proliferation, and augmented NK-mediated target cell killing. Our findings reveal a novel mode of cell-to-cell communication—allowing lymphocytes to extend the confines of their own proteome—which may moreover play an important role in natural tumor immunity

Acute and Chronic Effects of Particles on Hospital Admissions in New-England

Background: Many studies have reported significant associations between exposure to $PM_{2.5}$ and hospital admissions, but all have focused on the effects of short-term exposure. In addition all these studies have relied on a limited number of $PM_{2.5}$ monitors in their study regions, which introduces exposure error, and excludes rural and suburban populations from locations in which monitors are not available, reducing generalizability and potentially creating selection bias. Methods Using our novel prediction models for exposure combining land use regression with physical measurements (satellite aerosol optical depth) we investigated both the long and short term effects of $PM_{2.5}$ exposures on hospital admissions across New-England for all residents aged 65 and older. We performed separate Poisson regression analysis for each admission type: all respiratory, cardiovascular disease (CVD), stroke and diabetes. Daily admission counts in each zip code were regressed against long and short-term $PM_{2.5}$ exposure, temperature, socio-economic data and a spline of time to control for seasonal trends in baseline risk. Results: We observed associations between both short-term and long-term exposure to $PM_{2.5}$ and hospitalization for all of the outcomes examined. In example, for respiratory diseases, for every10-µg/m$^3$ increase in short-term $PM_{2.5}$ exposure there is a 0.70 percent increase in admissions (CI = 0.35 to 0.52) while concurrently for every10-µg/m$^3$ increase in long-term $PM_{2.5}$ exposure there is a 4.22 percent increase in admissions (CI = 1.06 to 4.75). Conclusions: As with mortality studies, chronic exposure to particles is associated with substantially larger increases in hospital admissions than acute exposure and both can be detected simultaneously using our exposure models

Cloud Coverage Acts as an Amplifier for Ecological Light Pollution in Urban Ecosystems

The diurnal cycle of light and dark is one of the strongest environmental factors for life on Earth. Many species in both terrestrial and aquatic ecosystems use the level of ambient light to regulate their metabolism, growth, and behavior. The sky glow caused by artificial lighting from urban areas disrupts this natural cycle, and has been shown to impact the behavior of organisms, even many kilometers away from the light sources. It could be hypothesized that factors that increase the luminance of the sky amplify the degree of this “ecological light pollution”. We show that cloud coverage dramatically amplifies the sky luminance, by a factor of 10.1 for one location inside of Berlin and by a factor of 2.8 at 32 km from the city center. We also show that inside of the city overcast nights are brighter than clear rural moonlit nights, by a factor of 4.1. These results have important implications for choronobiological and chronoecological studies in urban areas, where this amplification effect has previously not been considered