Growth resistance and resilience of mixed silver fir and Norway spruce forests in central Europe: Contrasting responses to mild and severe droughts.

Extreme droughts are expected to increase in frequency and severity in many regions of the world, threatening multiple ecosystem services provided by forests. Effective strategies to adapt forests to such droughts require comprehensive information on the effects and importance of the factors influencing forest resistance and resilience. We used a unique combination of inventory and dendrochronological data from a long-term (>30 years) silvicultural experiment in mixed silver fir and Norway spruce mountain forests along a temperature and precipitation gradient in southwestern Germany.

C-STABILITY an innovative modeling framework to leverage the continuous representation of organic matter.

The understanding of soil organic matter (SOM) dynamics has considerably advanced in recent years. It was previously assumed that most SOM consisted of recalcitrant compounds, whereas the emerging view considers SOM as a range of polymers continuously processed into smaller molecules by decomposer enzymes. Mainstreaming this new paradigm in current models is challenging because of their ill-adapted framework.

Cardiac function estimation from MRI using a heart model and data assimilation: advances and difficulties.

In this paper, we present a framework to estimate local ventricular myocardium contractility using clinical MRI, a heart model and data assimilation. First, we build a generic anatomical model of the ventricles including muscle fibre orientations and anatomical subdivisions. Then, this model is deformed to fit a clinical MRI, using a semi-automatic fuzzy segmentation, an affine registration method and a local deformable biomechanical model.

Fractionation analysis of the endosomal compartment during rat reticulocyte maturation.

A subcellular fractionation procedure was developed to isolate the different endosomal compartments present during reticulocyte maturation. After reticulocyte lysis and removal of excess haemoglobin by gel chromatography, membrane vesicles were separated over a discontinuous sucrose gradient (10-40%). Two fractions were isolated: P1 at the 25-35% sucrose interface and P2 at the 17-25% sucrose interface.

Exosomes released during reticulocyte maturation bind to fibronectin via integrin alpha4beta1.

Exosomes are vesicles formed in the endosomal compartment and released in the extracellular medium during reticulocyte maturation into erythrocytes. They have a clearing function because of their enrichment with some proteins known to decrease or disappear from the cell surface during maturation, e.g.

Lipid headgroup spacing and peptide penetration, but not peptide oligomerization, modulate peptide-induced fusion.

In this study, the mechanism by which an amphipathic negatively charged peptide consisting of 11 amino acids (WAE) induces fusion of liposomal phosphatidylcholine membranes is investigated. WAE-induced fusion, which only occurs when the peptide is covalently attached to the bilayer, shows a highly remarkable dependence on naturally occurring phosphatidylcholine species. The initial rate of fusion increased in the order 1-palmitoyl 2-arachidonoyl PC (PAPC) > 1-palmitoyl 2-oleoyl PC (POPC) > 1-stearoyl 2-oleoyl PC (SOPC) > dioleoyl PC (DOPC) > egg yolk PC.

Peptides and membrane fusion: towards an understanding of the molecular mechanism of protein-induced fusion.

Processes such as endo- or exocytosis, membrane recycling, fertilization and enveloped viruses infection require one or more critical membrane fusion reactions. A key feature in viral and cellular fusion phenomena is the involvement of specific fusion proteins. Among the few well-characterized fusion proteins are viral spike glycoproteins responsible for penetration of enveloped viruses into their host cells, and sperm proteins involved in sperm-egg fusion.

Exoenzyme S from P. aeruginosa ADP ribosylates rab4 and inhibits transferrin recycling in SLO-permeabilized reticulocytes.

ADP-ribosylation of rab proteins by exoenzyme S (Exo S) of P. aeruginosa was studied using reticulocytes. 14-3-3 protein, the eukaryotic cofactor that is obligatory for Exo S activity, was found in association with reticulocyte endocytic vesicles and exosomes, vesicles previously shown to be enriched with rab4.

Transferrin receptor functions as a signal-transduction molecule for its own recycling via increases in the internal Ca2+ concentration.

Transferrin binding to its receptor modulates transferrin receptor (Tf-R) recycling rates in several cells [Klausner, R. D., Van Renswoude, J.

Membrane anchorage brings about fusogenic properties in a short synthetic peptide.

The fusogenic properties of an amphipathic net-negative peptide (wae 11), consisting of 11 amino acid residues, were studied. We demonstrate that, whereas the free peptide displays no significant fusion activity, membrane fusion is strongly promoted when the peptide is anchored to a liposomal membrane. The fusion activity of the peptide appears to be independent of pH, and membrane merging is an essentially nonleaky process.

A rapid redistribution of transferrin receptors to the cell surface of L2C lymphocytes upon fixation of holoform transferrin.

The internalization and recycling kinetics of transferrin receptors in leukemic lymphocytes (L2C) differed in the absence or presence of ligand. At 37 degrees C in the absence of ligand, transferrin receptors were mainly distributed internaly. We demonstrated, using a sepharose-bead-Tf complex, the rapid recycling of unoccupied internal transferrin receptors was correlated with ligand binding to surface receptors.

A GTP-binding protein modulates a Ca2+ pump present in reticulocyte endocytic vesicles.

Rat reticulocytes were used to prepare endocytic vesicles to study calcium fluxes across the endosomal membrane. We used 45Ca2+, and found that endocytic vesicles from reticulocytes present a Ca(2+)-ATPase that pumps Ca2+ into the lumen of vesicles. This activity was sensitive to vanadate and the calmodulin antagonists: trifluoperazine and calmidazolium.

Quantitative comparison between aminophospholipid translocase activity in human erythrocytes and in K562 cells.

Spin-labeled phospholipids were used to determine the transbilayer movement of phospholipids in human erythrocytes, in K562 cells and in human neonatal red cells. The erythroleukemia cell line, K562, as well as human neonatal red cells, which are rich in reticulocytes, were considered as representative of human erythrocyte precursor cells. In the nucleated cells, the difference between outside-inside movement of aminophospholipids and that of phosphatidylcholine or sphingomyelin analogues allowed us to discriminate between lipid internalization due to aminophospholipid translocase activity and to endocytosis.

Effect of benzyl alcohol on phospholipid transverse mobility in human erythrocyte membrane.

The effect of benzyl alcohol on the transverse mobility and repartition of phospholipids in the human erythrocyte membrane was investigated using electron spin resonance and morphological modification of red blood cells. Transmembrane internalization rates and equilibrium distribution in red blood cells of short-chain spin-labeled phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine were strongly modified by treatment with 10-70 mM benzyl alcohol. A dual effect was observed: (a) at 4 degrees C and 37 degrees C there was an N-ethylmaleimide-sensitive, long lasting and fully reversible increase in the spin-labeled phosphatidylserine and phosphatidylethanolamine internalization rate; (b) at 37 degrees C, an enhancement of N-ethylmaleimide-insensitive fluxes of all the labeled phospholipids through the membrane occurred.

The influence of transferrin binding to L2C guinea pig leukemic lymphocytes on the endocytosis cycle kinetics of its receptor.

The parameters regulating the internalization and recycling of transferrin-specific receptors were determined in guinea pig leukemic B lymphocytes, in the absence or presence of ligand. We show that after the cells were purified, 45-56% of the total receptors were on the cell surface. In the absence of transferrin, unoccupied receptors are quickly internalized (rate constant, 0.

Structural abnormality in LDL from diabetic patients as revealed by resonance Raman spectroscopy.

Resonance Raman spectra of low-density lipoprotein (LDL) isolated from the plasma of diabetic patients (age range 13-77 yr, mean age 37 yr) and age- and sex-matched control subjects were recorded in the 1000- to 1600-cm-1 region as a function of temperature (0-50 degrees C). Both nondiabetic and diabetic LDL yield spectra characterized by two major bands near 1160 and 1530 cm-1 due to the carotenoid component of lipoproteins. The relative intensity of 1530- and 1160-cm-1 bands, assigned to -C = C- and = C-C = stretchings, respectively, i.

Effects of benzyl alcohol on transferrin and low density lipoprotein receptor mediated endocytosis in leukemic guinea pig B lymphocytes.

We demonstrated that benzyl alcohol, a neutral local anesthetic drug, inhibits the uptake and degradation of lowdensity lipoprotein and endocytosis of transferrin receptors of guinea pig leukemic B lymphocytes (L2C). This inhibition is very rapid, concentration dependant and reversible by simple washing. Membrane fluidity of the living cells is also modified.

Asymmetric distribution of phospholipids in the membrane of vesicles released during in vitro maturation of guinea pig reticulocytes: evidence precluding a role for "aminophospholipid translocase".

Guinea pig reticulocytes lose their transferrin (Tf) binding activity during maturation, in the form of vesicles (exosomes) released into the extracellular medium. Vesicles were prepared from cultures of reticulocytes to study the possible externalization of a particular membrane-associated activity, i.e.

Modifications of LDL-receptor-mediated endocytosis rates in CEM lymphoblastic cells grown in lipoprotein-depleted fetal calf serum.

The efficiency of supplying cholesterol by the LDL endocytic pathway of lymphoblastic T CEM cells was compared when incubated in the presence of either fetal calf serum (FCS) or lipoprotein-depleted fetal calf serum (LDFCS). In the presence of FCS, there were 8600 +/- 2000 LDL receptors/cell with a Kd of (2.2 +/- 0.

Evidence for bidirectional transverse diffusion of spin-labeled phospholipids in the plasma membrane of guinea pig blood cells.

The distribution and transverse diffusion kinetics of four spin-labeled phospholipid analogues (two with choline heads: phosphatidylcholine (PC) and sphingomyelin (SM); two with amino heads: phosphatidylserine (PS) and phosphatidylethanolamine (PE) were studied in the plasma membrane of guinea pig blood cells: erythrocytes, reticulocytes, and leukemic lymphocytes. Nitroxide reduction by the internal content of the cells was used as an indicator to determine the phospholipids that penetrated the cells. The reduction rates were in the order, PS greater than PE greater than PC greater than SM in all cells.

Regulation of cholesterol biosynthesis at a stage after the 3-hydroxy-3-methylglutaryl-CoA reductase step, in normal and leukemic (L2C) guinea pig lymphocytes.

Cholesterogenic activity in normal and leukemic guinea pig lymphocytes was measured by incorporation of labeled sodium acetate into cholesterol, after separation from other labeled metabolites. Our study is in agreement with the large difference previously found between the two kinds of cells at the 3-hydroxy-3-methylglutaryl-CoA reductase step, but it also shows that the difference is not as great as described earlier, when expressed in terms of the final product, cholesterol. This is mainly due to differences in the analytical methods.

The influence of coupling transferrin to liposomes or minibeads on its uptake and fate in leukemic L2C cells.

Coupling transferrin to liposomes or minibeads did not affect its uptake by L2C lymphocytes via the Tf specific receptors. The uptake kinetics of Tf conjugated with particles about 50 nm in diameter was as rapid as in the case of native Tf, and the receptors were recycled with a similar turnover time (about 15 min). Contrary to the generally accepted scheme, we found some Tf degradation provoked by cellular uptake.

Internalization of low-density-lipoprotein-specific receptors in leukemic guinea pig lymphocytes.

Leukemic guinea pig lymphocytes (L2C) have ten times as many low-density lipoprotein (LDL) receptors as healthy lymphocytes, but LDL accounts for only 38% of the cholesterol in L2C cells, compared to more than 95% in normal cells. Our data show that LDL fails to regulate cholesterol biosynthesis and that there is a defect in LDL internalization and receptor turnover in L2C cells. We also demonstrate that the degradation of LDL is not a limiting process.

Thiolation of low-density lipoproteins and their interaction with L2C leukemic lymphocytes.

We present here, a new method for coupling sulfhydryl groups (SH) to low-density lipoprotein (LDL) surface. This method uses homocysteine thiolactone (HCTL) which reacts with lysine residues in a very mild manner, and permits the selection of the number of SH bound per LDL. Under our experimental conditions (8 SH/LDL), the affinity of thiolated LDL for the specific receptors and their further internalization by L2C lymphocytes are preserved.