Lipid phase transitions and phase diagrams. II. Mictures involving lipids.
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Adenosine Triphosphatases↗
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[Transitional phases in human development (author's transl)].
Transitional phases in the course of Human Development which divide the course of development into different steps, have been searched far from the old. These transitional phases are very often seen to be at least initiated by physical resp. biological changes and accompanied by psychological or mental changes. The results of the Life-span-Psychology and especially the findings of the biographical studies of our Psychological Institue, which were started in the Fifties, shows that there are Transitional phases which neither related to biological change nor to role change nor to the change from the one decade of life to the other one. Much more important are very personal experiences which very often lead to a reorientation. - These findings are in agreement with cognitive theory of personality, according to which human behavior is dependent more on the situation as perceived, than on the stimulus-pattern contigent with this situation. Therefore the study of these situational concepts of individual is relevant for the Life-span-Psychology and specially for the analysis of transitional phases.
Aging↗
Role of lipids in the Neurospora crassa membrane. I. Influence of fatty acid composition on membrane lipid phase transitions.
The relationship between lipid composition and phase transition was investigated by differential scanning calorimetry for intact and membrane phospholipid extracts of wild-type (w/t) and the cel-(Tw 40) mutant of Neurospora crassa. The cel-(Tw 40) mutant (grown on minimal, sucrose medium supplemented with Tween 40 at approximately 34 degrees C) had approximately twice the saturated fatty acid content of w/t organisms grown at approximately 22 degrees C. The gel-liquid crystal phase transitions of ergosterol-free extracts derived from w/t and cel-(Tw 40) occur at -31 and -11 degrees C, respectively. The heats of transition (delta H) of these extracts were 1 and 13 cal/g, respectively. The addition of ergosterol (the predominant sterol in Neurospora) to the phospholipid extracts decreased the observed heats of transition, but did not alter the transition temperature. Intact Neurospora, whether w/t or cal-(Tw 40) did not manifest similar gel-liquid crystal phase transitions in the differential scanning calorimeter. However, an endothermic peak at approximately 30 degrees C was observed in intact cells and extracted phospholipids of both w/t and cel-(Tw 40) organisms. This peak was insensitive to the addition of ergosterol, had a low heat content (delta H congruent to 1 cal/g), and was reversible.
Calorimetry, Differential Scanning↗
Cellular modifiers of TDP-43 phase transition and cytoplasmic aggregation.
RNA-binding protein TAR DNA-binding protein 43 (TDP-43) can form liquid-like nuclear assemblies whose phase behavior is thought to influence its aggregation propensity and neurotoxic activity. The mechanisms that govern the liquid-to-solid phase transition of TDP-43 remain poorly defined. Here, we combined chemical and genome-wide genetic screens to identify cellular factors that modulate the phase behavior of an RNA-binding-defective TDP-43 mutant. Our screens uncovered multiple cellular processes, including RNA splicing, protein translation, proteostasis imbalance, and nuclear export as TDP-43 phase regulators. We also developed a semi-permeabilized cell system that partially recapitulates the TDP-43 phase transition in vitro, and showed that nuclear export inhibition reshapes the nuclear environment to favor RNA-dependent liquid-liquid phase separation (LLPS) of TDP-43, which mitigates its aggregation. Nuclear export inhibition in a brain organoid model bearing an ALS-associated mutation reduces pathogenic phospho-TDP-43 accumulation. These findings identify multiple modulators of TDP-43 phase transitions in a sensitized model system and establish a framework for further dissecting the link between nuclear transport and TDP-43 phase dynamics.
DNA-Binding Proteins↗
Depression of phase-transition temperature in a model cell membrane by local anesthetics.
Disordering, fluidizing and dilating effects of anesthetics upon cell membranes are well recognized. The fluidization can be precisely measured with phospholipid membranes. When phospholipids are dispersed in water, they form globules of bilayer structure. These model membranes undergo transition between crystalline (ordered and less fluid) and liquid crystalline (less ordered and fluid) phases according to the temperature, the degree of packing of each molecule, and the chemical environment. The phase transition occurs in a cooperative fashion and the turbidity of the dispersion changes abruptly, clear in liquid crystalline phase and turbid in crystalline phase. The present study was undertaken to quantitate the fluidizing effects of local anesthetics on dipalmitoyl lecithin (DPL) bilayer dispersion by measuring the turbidity change. Tetracaine, bupivacaine, lidocaine, and procaine were studied. They all depressed the phase-transition temperature. The binding of the drugs to the model membrane followed unsaturable kinetics, and the pH titration curve showed that only uncharged molecules were active. The freezing point depression was analyzed according to the Van't Hoff model. From this model, the partition coefficients of the uncharged molecules between DPL and water were estimated: lidocaine 76, procaine 159, bupivacaine 812, and tetracaine 1,405. The concentration of local anesthetics in the DPL phase needed to decrease the phase-transition temperature 1 degree C showed a constant value of 0.132 M. The concentration of local anesthetics in the DPL phase is a function of pH, partition coefficient, and volume ratio between the DPL and aqueous phases. The normalized values of the fluidizing action of these drugs at physiologic conditions correlated well with their nerve-blocking potencies. The present results indicate that the uncharged molecules fluidize the lecithin membrane by unsaturable nonspecific binding. The possible effect of the charged molecules upon the fluidity of natural membranes remains to be established.
Anesthetics, Local↗
Ionic influences on the phase transition of dipalmitoylphosphatidylserine.
The ionization and phase behavior of 1,2-dipalmitoyl-sn-glycero-3-phosphoserine have been investigated under a variety of condtions by several different methods. As measured by turbidity changes, the temperature of the crystal-liquid crystal phase transition of this lipid is influenced by pH and mono- and divalent cation concentrations. The pH-transition temperature curve is congruent with the curve relating temperature to the degree of ionization of the carboxyl group of the crystalline form. The transition temperature falls from an upper plateau of 72 degrees C at low pH values, where the carboxyl group is fully protonated, to a lower plateau of 55 degrees C at high pH values, where this group is fully ionized. The apparent pK (pH at 50% ionization) of the crystalline form shifts from 6.0 to 4.6 to 3.7 with an increase of NaCl concentration from 10(-3) to 0.1 to l.0 M, respectively. These observations are in accord with a simple theoretical analysis that utilizes diffuse double layer theory and the influence of surface potential on surface concentration of protons. In qualitative terms, an increase in electrolyte concentration reduces the surface potential, the result of which is a diminution of the surface-bulk pH difference and a lowering of the apparent pK. Assuming an area of 50 A2/molecule, the intrinsic pKa (apparent pK corrected for surface pH) of the carboxyl group is 2.7. A 1000-fold change of NaCl concentration produces a very large change in surface potential without influencing the transition temperature of the ionized form of the lipid.
Calorimetry↗
Phase transition in charged lipid membranes.
Experimental results on the effect of electrostatics on bilayer phase transitions are compared with corresponding data for monolayers and the predictions of electrical double layer theory. The two substantial conclusions which emerge are that: (i) double layer theory based on a continuous surface charge distribution cannot explain all the relevant data, a situation which may be improved by taking into account the discrete nature of the surface charge distribution; (ii) the crystal - liquid crystal phase transition of charged bilayer membranes is always a continuous one which takes place through an intermediate state consisting of both fluid and frozen domains.
Electricity↗
Effects of proteins on thermotropic phase transitions of phospholipid membranes.
A variety of proteins have been studied for their ability to interact and alter the thermotropic properties of phospholipid bilayer membranes as detected by differential scanning calorimeter. The proteins studied included: basic myelin protein (A1 protein), cytochrome c, major apoprotein of myelin proteolipid (N-2 apoprotein), gramicidin A, polylysine, ribonuclease and hemoglobin. The lipids used for the interactions were dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylglycerol. The interactions were grouped in three catagories each having very different effects on the phospholipid phase transition from solid to liquid crystalline. The calorimetric studies were also correlated with data from vesicle permeability and monolayer expansion. Ribonuclease and polylysine which exemplify group 1 interactions, show strong dependence on electrostatic binding. Their effects on lipid bilayers include an increase in the enthalpy of transition (deltaH) accompanied by either an increase or no change in the temperature of transition (Tc). In addition, they show minimal effects on vesicle permeability and monolayer expansion. It was concluded that these interactions represent simple surface binding of the protein on the lipid bilayer without penetration into the hydrocarbon region. Cytochrome c and A1 protein, which exemplify group 2 interactions, also show a strong dependence on the presence of net negative charges on the lipid bilayers for their binding. In contrast to the first group, however, they induce a drastic decrease in both Tc and deltaH of the lipid phase transition. Furthermore, they induce a large increase in the permeability of vesicles and a substantial expansion in area of closely packed monolayers at the air-water interface. It was concluded that group 2 interactions represent surface binding followed by partial penetration and/or deformation of the bilayer. Group 3 interactions, shown by proteolipid apoprotein and gramicidin A, were primarily non-polar in character, not requiring electrostatic charges and not inhibited by salt and pH changes. They had no appreciable effect on the Tc but did induce a linear decrease in the magnitude of the deltaH, proportional to the percentage of protein by weight. Membranes containing 50% proteolipid protein still exhibited a thermotropic transition with a deltaH one half that of the pure lipid, and only a small diminution of the size of the cooperative unit. It was concluded that in this case the protein was embedded within the bilayer, associating with a limited number of molecules via non-polar interactions, while the rest of the bilayer was largely unperturbed.
Animals↗
Lateral and vertical displacement of integral membrane proteins during lipid phase transition in Anacystis nidulans.
Alterations in membrane structure as a result of lipid phase transitions have been studied in Anacystis nidulans, a blue-green alga. Cells grown at 38 degrees C were subjected to temperature transitions of 38 degrees C leads to 21 degrees C and 38 degrees C leads to 10 degrees C, previously shown to produce substantial changes in photosynthetic activities, and examined by freeze-fracture electron microscopy. As a result of these treatments, large particle-free regions appeared on the fracture faces of both the plasma and thylakoid membranes. Particle density measurements suggest that the displacement of the integral membrane protein complexes occurs in both lateral and vertical directions. Returning the cells to 38 degrees C resulted in the restoration of normal membrane morphology, indicating that the proteins were not lost from the membrane. Such displacement of the integral membrane protein complexes could contribute significantly to the temperature-dependent alterations in the functional activity of membrane-bound enzymatic complexes.
Cell Membrane↗
Phase Transition of Wax Enabling CRISPR Diagnostics for Automatic At-Home Testing of Multiple Sexually Transmitted Infection Pathogens.
Sexually transmitted infections (STIs) significantly impact women's reproductive health. Rapid, sensitive, and affordable detection of these pathogens is essential, especially for home-based self-testing, which is crucial for individuals who prioritize privacy or live in areas with limited access to healthcare services. Herein, an automated diagnostic system called Wax-CRISPR has been designed specifically for at-home testing of multiple STIs. This system employs a unique strategy by using the solid-to-liquid phase transition of wax to sequentially isolate and mix recombinase polymerase amplification (RPA) and CRISPR assays in a microfluidic chip. By incorporating a home-built controlling system, Wax-CRISPR achieves true one-pot multiplexed detection. The system can simultaneously detect six common critical gynecological pathogens (CT, MG, UU, NG, HPV 16, and HPV 18) within 30 min, with a detection limit reaching 10-18 M. Clinical evaluation demonstrates that the system achieves a sensitivity of 96.8% and a specificity of 97.3% across 100 clinical samples. Importantly, eight randomly recruited untrained operators performe a double-blinded test and successfully identified the STI targets in 33 clinical samples. This wax-transition-based one-pot CRISPR assay offers advantages such as low-cost, high-stability, and user-friendliness, making it a useful platform for at-home or field-based testing of multiple pathogen infections.
Sexually Transmitted Diseases↗
Nuclear export modulates TDP-43 phase transition and cytoplasmic aggregation.
RNA-binding protein TAR DNA-binding protein 43 (TDP-43) can form liquid-like, nuclear assemblies whose phase behavior may influence its aggregation propensity and neurotoxic activity. The mechanism(s) that modulates the transition of TDP-43 from a liquid to solid phase is poorly defined. Here we combine chemical and genome-wide genetic screenings to identify cellular factors that modulate the phase behavior of an RNA-binding defective TDP-43 mutant that mimics an Amyotrophic Lateral Sclerosis (ALS)-associated variant. Our screens uncover multiple cellular processes including RNA splicing, protein translation, proteostasis imbalance and nuclear export as TDP-43 phase regulators. Importantly, TDP-43 phase transition can be dynamically recapitulated in vitro in a semi-permeabilized cell system, which reveals that the inhibition of nuclear export reshapes the nuclear environment in favor of an RNA-dependent TDP-43 liquid-liquid phase separation (LLPS) state, which mitigates cytoplasmic TDP-43 aggregation. We validated this mechanism in a brain organoid model bearing an ALS-associated mutation, showing that nuclear export deficiency can limit pathogenic phospho-TDP-43 accumulation. These findings establish nuclear export as a key regulator of TDP-43 phase transitions and define a mechanistic framework that links altered nuclear transport and phase dynamics to TDP-43 aggregation potential.
Amyotrophic Lateral Sclerosis (ALS)↗
[Sedimentation rate of erythrocytes as an indicator for phase transitions in the membrane].
Temperature dependent sedimentation measurements with human erythrocytes showed that the sedimentation velocity at 21 degrees C in Krebs-Ringer-solution, pH 7.4, has a peak shaped minimum [1]. In further investigations it could be shown that a phase transition of membrane lipids is the main reason of this peak. Membrane proteins influence the sedimentation behavior of the erythrocytes only by changing the interaction with membrane lipids.
Blood Sedimentation↗
Cross-feeding percolation phase transitions of intercellular metabolic networks.
Intercellular cross-talk is essential for the adaptation capabilities of populations of cells. While direct diffusion-driven cell-to-cell exchanges are difficult to map, current nanotechnology enables one to probe single-cell exchanges with the medium. We introduce a mathematical method to reconstruct the dynamic unfolding of intercellular exchange networks from these data, applying it to an experimental coculture system. The exchange network, initially dense, progressively fragments into small disconnected clusters. To explain these dynamics, we develop a maximum-entropy multicellular metabolic model with diffusion-driven exchanges. The model predicts a transition from a dense network to a sparse one as nutrient consumption shifts. We characterize this crossover both numerically, revealing a power-law decay in the cluster-size distribution, and analytically, by connecting to percolation theory. Comparison with data suggests that populations evolve toward the sparse phase by remaining near the crossover. These findings offer insights into the collective organization driving the adaptive dynamics of cell populations.
Metabolic Networks and Pathways↗
Coronary venous flow and O2 saturation during transitional phases between various cardiac rates.
The instantaneous and continuous interrelationship between coronary blood flow and coronary venous O2 saturation was determined during transient periods following abrupt rate change in the electrically paced canine heart. Through a catheter in the coronary sinus, O2 saturation was continuously monitored using a fibreoptics technique, and venous flow was measured with an electromagnetic flowmeter. Various patterns of change in flow and O2 saturation were observed depending both on the absolute values of the cardiac rates as well as on the relative difference between them during changes from one rate to another. Whereas elevation of coronary flow was monophasic when the magnitude of heart rate change was below 75 beats per minute, a drop in flow was observed preceeding its elevation when the difference was greater. At high rates further increase in rate caused either no alteration or led to a monophasic drop in flow during the transitional period. Changes in O2 saturation were observed only when heart rate difference exceeded 60 beats per minute. Between 60-90 beats per minute O2 saturation remained steady except during the transient rate elevation, ending in a lower steady state O2 saturation. The results indicate that both O2 saturation and coronary flow change with heart rate initially because of mechanical consequence of the increased rate on the myocardium, and later according to its new metabolic needs also manifested by changed O2 extraction.
Animals↗
Comparison of thymidine, fluorodeoxyuridine, hydroxyurea, and methotrexate blocking at the G1/S phase transition of the cell cycle, studied by replication patterns.
Cultures of human amniotic fluid cells and fibroblasts were temporarily blocked by the replication inhibitors thymidine (dT) surplus, fluorodeoxyuridine (FdU), hydroxyurea (HU), or methotrexate uridine (MU). The respective arresting point at G1-S transition and the homogeneity of the blocked cell population were determined by means of BrdU replication patterns. Most variation of patterns were found after HU. After MU, cells were arrested before the onset of replication, while with dT surplus of FdU an arresting point in early S seemed more likely.
Amniotic Fluid↗
Influence of surface potentials on the mitochondrial H+ pump and on lipid-phase transitions.
It has been shown that the surface potential of lipid membranes, as well as of mitochondria, can be shifted more positive by absorption of alkylbiguanides. Both phospholipid vesicles and natural membranes respond in an analogous way to this shift. Ion activities at the immediate membrane surface are influenced by sign and magnitude of the surface charge. Corresponding effects on ion transport and on fluorescence-probe binding can be observed. The mitochondrial H+ pump is inhibited when the surface charge is shifted more positive. In contrast,the absolute charge density determines the temperature of the ordered-fluid transition. The latter is increased by biguanides, suggesting that the membrane is rendered more rigid. The experiments make obvious that physical relations derived from model systems apply equally well to lipid-containing natural membranes.
Aerobiosis↗
Sphingomyelin phase transition in the sheep erythrocyte membrane.
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Animals↗
Phase transitions and coupling in energy transducing membranes.
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Adenosine Triphosphatases↗