Functional consequences of sphingomyelinase-induced changes in erythrocyte membrane structure.

Inflammation enhances the secretion of sphingomyelinases (SMases). SMases catalyze the hydrolysis of sphingomyelin into phosphocholine and ceramide. In erythrocytes, ceramide formation leads to exposure of the removal signal phosphatidylserine (PS), creating a potential link between SMase activity and anemia of inflammation. Therefore, we studied the effects of SMase on various pathophysiologically relevant parameters of erythrocyte homeostasis. Time-lapse confocal microscopy revealed a SMase-induced transition from the discoid to a spherical shape, followed by PS exposure, and finally loss of cytoplasmic content. Also, SMase treatment resulted in ceramide-associated alterations in membrane-cytoskeleton interactions and membrane organization, including microdomain formation. Furthermore, we observed increases in membrane fragility, vesiculation and invagination, and large protein clusters. These changes were associated with enhanced erythrocyte retention in a spleen-mimicking model. Erythrocyte storage under blood bank conditions and during physiological aging increased the sensitivity to SMase. A low SMase activity already induced morphological and structural changes, demonstrating the potential of SMase to disturb erythrocyte homeostasis. Our analyses provide a comprehensive picture in which ceramide-induced changes in membrane microdomain organization disrupt the membrane-cytoskeleton interaction and membrane integrity, leading to vesiculation, reduced deformability, and finally loss of erythrocyte content. Understanding these processes is highly relevant for understanding anemia during chronic inflammation, especially in critically ill patients receiving blood transfusions

Subclinical giant cell arteritis in polymyalgia rheumatica: Concurrent conditions or a common spectrum of inflammatory diseases?

Giant cell arteritis (GCA) and polymyalgia rheumatica (PMR) are common conditions in older adults. Their clinical connection has been recognized over time, with many patients experiencing both conditions separately, simultaneously or in temporal sequence to each other. Early GCA detection is essential to prevent vascular damage, but identifying subclinical GCA in PMR patients remains a challenge and routine screening is not standard practice. Subclinical GCA prevalence in newly diagnosed PMR patients ranges from 23 to 29%, depending on the screening method. Vessel wall imaging and temporal artery biopsy can detect subclinical GCA. Epidemiology and trigger factors show similarities between the two conditions, but PMR is more common than GCA. Genetic and pathogenesis studies reveal shared inflammatory mechanisms involving dendritic cells, pro-inflammatory macrophages, and an IL-6 signature. However, the inflammatory infiltrates differ, with extensive T cell infiltrates seen in GCA while PMR shows an incomplete profile of T cell and macrophage-derived cytokines. Glucocorticoid treatment is effective for both conditions, but the steroid requirements vary. PMR overall mortality might be similar to the general population, while GCA patients with aortic inflammatory aneurysms face increased mortality risk. The GCA-PMR association warrants further research. Considering their kinship, recently the term GCA-PMR Spectrum Disease (GPSD) has been proposed

A quantitative study on the growth variability of tumour cell clones in vitro

Objectives: In this study, we quantify the growth variability of tumour cell clones from a human leukemia cell line. Materials and methods: We have used microplate spectrophotometry to measure the growth kinetics of hundreds of individual cell clones from the Molt3 cell line. The growth rate of each clonal population has been estimated by fitting experimental data with the logistic equation. Results: The growth rates were observed to vary among different clones. Up to six clones with a growth rate above or below the mean growth rate of the parent population were further cloned and the growth rates of their offsprings were measured. The distribution of the growth rates of the subclones did not significantly differ from that of the parent population thus suggesting that growth variability has an epigenetic origin. To explain the observed distributions of clonal growth rates we have developed a probabilistic model assuming that the fluctuations in the number of mitochondria through successive cell cycles are the leading cause of growth variability. For fitting purposes, we have estimated experimentally by flow cytometry the maximum average number of mitochondria in Molt3 cells. The model fits nicely the observed distributions of growth rates, however, cells in which the mitochondria were rendered non functional (rho-0 cells) showed only a 30% reduction in the clonal growth variability with respect to normal cells. Conclusions: A tumor cell population is a dynamic ensemble of clones with highly variable growth rate. At least part of this variability is due to fluctuations in the number of mitochondria.Comment: 31 pages, 5 figure

Deciphering the components of regional net ecosystem fluxes following a bottom-up approach for the Iberian Peninsula

Quantification of ecosystem carbon pools is a fundamental requirement for estimating carbon fluxes and for addressing the dynamics and responses of the terrestrial carbon cycle to environmental drivers. The initial estimates of carbon pools in terrestrial carbon cycle models often rely on the ecosystem steady state assumption, leading to initial equilibrium conditions. In this study, we investigate how trends and inter-annual variability of net ecosystem fluxes are affected by initial non-steady state conditions. Further, we examine how modeled ecosystem responses induced exclusively by the model drivers can be separated from the initial conditions. For this, the Carnegie-Ames-Stanford Approach (CASA) model is optimized at set of European eddy covariance sites, which support the parameterization of regional simulations of ecosystem fluxes for the Iberian Peninsula, between 1982 and 2006. <br><br> The presented analysis stands on a credible model performance for a set of sites, that represent generally well the plant functional types and selected descriptors of climate and phenology present in the Iberian region – except for a limited Northwestern area. The effects of initial conditions on inter-annual variability and on trends, results mostly from the recovery of pools to equilibrium conditions; which control most of the inter-annual variability (IAV) and both the magnitude and sign of most of the trends. However, by removing the time series of pure model recovery from the time series of the overall fluxes, we are able to retrieve estimates of inter-annual variability and trends in net ecosystem fluxes that are quasi-independent from the initial conditions. This approach reduced the sensitivity of the net fluxes to initial conditions from 47% and 174% to −3% and 7%, for strong initial sink and source conditions, respectively. <br><br> With the aim to identify and improve understanding of the component fluxes that drive the observed trends, the net ecosystem production (NEP) trends are decomposed into net primary production (NPP) and heterotrophic respiration (<i>R</i><sub>H</sub>) trends. The majority (~97%) of the positive trends in NEP is observed in regions where both NPP and <i>R</i><sub>H</sub> fluxes show significant increases, although the magnitude of NPP trends is higher. Analogously, ~83% of the negative trends in NEP are also associated with negative trends in NPP. The spatial patterns of NPP trends are mainly explained by the trends in <i>f</i>APAR (<i>r</i>=0.79) and are only marginally explained by trends in temperature and water stress scalars (<i>r</i>=0.10 and <i>r</i>=0.25, respectively). Further, we observe the significant role of substrate availability (<i>r</i>=0.25) and temperature (<i>r</i>=0.23) in explaining the spatial patterns of trends in <i>R</i><sub>H</sub>. These results highlight the role of primary production in driving ecosystem fluxes. <br><br> Overall, our study illustrates an approach for removing the confounding effects of initial conditions and emphasizes the need to decompose the ecosystem fluxes into its components and drivers for more mechanistic interpretations of modeling results. We expect that our results are not only specific for the CASA model since it incorporates concepts of ecosystem functioning and modeling assumptions common to biogeochemical models. A direct implication of these results is the ability of this approach to detect climate and phenology induced trends regardless of the initial conditions

Bortezomib-induced muscle toxicity in multiple myeloma

Multiple myeloma (MM) accounts for ~13% of all hematologic malignancies. Bortezomib treatment is effective in MM, but can be complicated with neurological side effects. We describe a patient with symptomaticMM who had a reversible metabolic myopathy associated with bortezomib administration and pathologically characterized by excessive storage of lipid droplets together with mitochondrial abnormalities. In a single-center prospective study, 14 out of 24 patients with symptomatic MM were treated with bortezomib and, among these, 7 developed muscular signs and/or symptoms. The myopathy was characterized by a proximal muscle weakness involving lower limbs and was an early complication. Complete resolution of muscle weakness occurred after treatment discontinuation. Conversely, none of the patients who received a treatment without bortezomib developed muscular symptoms. Experimental studies demonstrate that in primary human myoblasts bortezomib at low concentrations leads to excessive storage of lipid droplets together with structural mitochondrial abnormalities, recapitulating the pathologic findings observed in patient's muscle. Our data suggest that patients treated with bortezomib should be monitored for muscular signs and/or symptoms and muscle weakness should alert the clinician to the possibility of myopathy. Bortezomib-induced metabolic myopathy is a potentially reversible entity with important implications for management and treatment of patients with MM

Performance of a novel sieving matrix of Poly(vinyl alcohol)ACRYLAMIDE copolymer in elecrophoretic separations of high molecular weight proteins from red cell membrane.

The analysis of high molecular weight (HMW) proteins from complex mixtures is still a challenge in proteomics. This work introduces a novel hydrogel obtained by the copolymerization of an allyl-PVA derivative with acrylamide and bisacrylamide and applies this matrix to the electrophoretic separation of HMW proteins. By inducing gelation of polyacrylamide in the presence of variable amounts of allyl-PVA, it is possible to control and vary the average gel porosity. This gel is easy to produce and handle and offers the advantage of being highly mechanically resistant and macroporous. The new matrix was tested in mono-dimensional separations of complex protein mixtures extracted from red cell membranes with different detergents. The improved performance of this macroporous matrix allowed to identify new proteins by MS and immunoblot analysis using specific antibodies. In particular, the resolution of proteins ranging in size between 97 and 279 kDa was greatly improved here compared to standard polyacrylamide gels, suggesting that this matrix can be a useful tool in routine analysis of HMW proteins in cell biology

Analyzing the causes and spatial pattern of the European 2003 carbon flux anomaly using seven models

Globally, the year 2003 is associated with one of the largest atmospheric CO<sub>2</sub> rises on record. In the same year, Europe experienced an anomalously strong flux of CO<sub>2</sub> from the land to the atmosphere associated with an exceptionally dry and hot summer in Western and Central Europe. In this study we analyze the magnitude of this carbon flux anomaly and key driving ecosystem processes using simulations of seven terrestrial ecosystem models of different complexity and types (process-oriented and diagnostic). We address the following questions: (1) how large were deviations in the net European carbon flux in 2003 relative to a short-term baseline (1998–2002) and to longer-term variations in annual fluxes (1980 to 2005), (2) which European regions exhibited the largest changes in carbon fluxes during the growing season 2003, and (3) which ecosystem processes controlled the carbon balance anomaly . <br><br> In most models the prominence of 2003 anomaly in carbon fluxes declined with lengthening of the reference period from one year to 16 years. The 2003 anomaly for annual net carbon fluxes ranged between 0.35 and –0.63 Pg C for a reference period of one year and between 0.17 and –0.37 Pg C for a reference period of 16 years for the whole Europe. <br><br> In Western and Central Europe, the anomaly in simulated net ecosystem productivity (NEP) over the growing season in 2003 was outside the 1σ variance bound of the carbon flux anomalies for 1980–2005 in all models. The estimated anomaly in net carbon flux ranged between –42 and –158 Tg C for Western Europe and between 24 and –129 Tg C for Central Europe depending on the model used. All models responded to a dipole pattern of the climate anomaly in 2003. In Western and Central Europe NEP was reduced due to heat and drought. In contrast, lower than normal temperatures and higher air humidity decreased NEP over Northeastern Europe. While models agree on the sign of changes in simulated NEP and gross primary productivity in 2003 over Western and Central Europe, models diverge in the estimates of anomalies in ecosystem respiration. Except for two process models which simulate respiration increase, most models simulated a decrease in ecosystem respiration in 2003. The diagnostic models showed a weaker decrease in ecosystem respiration than the process-oriented models. Based on the multi-model simulations we estimated the total carbon flux anomaly over the 2003 growing season in Europe to range between –0.02 and –0.27 Pg C relative to the net carbon flux in 1998–2002

Analyzing the causes and spatial pattern of the European 2003 carbon flux anomaly in Europe using seven models

International audienceGlobally, the year 2003 is associated with one of the largest atmospheric CO2 rises on record. In the same year, Europe experienced an anomalously strong flux of CO2 from the land to the atmosphere associated with an exceptionally dry and hot summer in Western and Central Europe. In this study we analyze the magnitude of this carbon flux anomaly and key driving ecosystem processes using simulations of seven terrestrial ecosystem models of different complexity and types (process-oriented and diagnostic). We address the following questions: (1) how large were deviations in the net European carbon flux in 2003 relative to a short-term baseline (1998–2002) and to longer-term variations in annual fluxes (1980 to 2005), (2) which regions exhibited the largest shift in carbon fluxes during the growing season 2003, and (3) which processes controlled the carbon balance anomaly . In Western and Central Europe, the anomaly in net ecosystem productivity (NEP) over growing season 2003 was outside the 1s bound of the carbon flux anomalies for 1980–2005. The estimated growing season anomaly ranged between –29 and –196 Tg C for Western Europe and between 13 and –94 Tg C for Central Europe depending on the model used. All models responded to a dipole pattern of the climate anomaly in 2003. In Western and Central Europe NEP was reduced due to heat and drought. Over Western Russia NEP was decreased in response to lower than normal temperatures and high precipitation. While models agree on changes in simulated NEP and gross primary productivity anomalies in 2003 over Western and Central Europe, models diverge in the estimates of anomalies in ecosystem respiration. Except for two process models which simulate respiration increase, most models simulated a decrease in ecosystem respiration in 2003. The diagnostic models showed a weaker decrease in ecosystem respiration than the process-oriented models. Based on the multi-model simulations we estimated the total carbon flux anomaly over the 2003 growing season in Europe to range between –0.02 and –0.27 Pg C relative to the net flux in 1998–2002