The dynamics and role of sphingolipids in eukaryotic organisms upon thermal adaptation

All living beings have an optimal temperature for growth and survival. With the advancement of global warming, the search for understanding adaptive processes to climate changes has gained prominence. In this context, all living beings monitor the external temperature and develop adaptive responses to thermal variations. These responses ultimately change the functioning of the cell and affect the most diverse structures and processes. One of the first structures to detect thermal variations is the plasma membrane, whose constitution allows triggering of intracellular signals that assist in the response to temperature stress. Although studies on this topic have been conducted, the underlying mechanisms of recognizing thermal changes and modifying cellular functioning to adapt to this condition are not fully understood. Recently, many reports have indicated the participation of sphingolipids (SLs), major components of the plasma membrane, in the regulation of the thermal stress response. SLs can structurally reinforce the membrane or/and send signals intracellularly to control numerous cellular processes, such as apoptosis, cytoskeleton polarization, cell cycle arresting and fungal virulence. In this review, we discuss how SLs synthesis changes during both heat and cold stresses, focusing on fungi, plants, animals and human cells. The role of lysophospholipids is also discussed.     

An Energy Analysis of Extracellular Hyperthermia

The classical hyperthermia effect is based on well‐focused energy absorption targeting the malignant tissue. The treatment temperature has been considered as the main technical parameter. There are discussions about the mechanism and control of the process because of some doubts about the micro‐mechanisms. The main idea of the extracellular hyperthermia is to heat up the targeted tissue by means of electric field, keeping the energy absorption in the extracellular liquid. This produces a temperature gradient and connected heat flow through the cell membrane, which initializes numerous nonequilibrium thermal microprocesses to destroy the cell membrane. Furthermore, before the heat shock activates the intracellular heat shock protein (HSP) mechanisms, thecell membrane has been already compromised, therefore the HSP synthesis in the cells starts secondarily only after the membrane damage. The process could explain why the nonuniform and basically unsatisfactorily high temperature locoregional hyperthermia could be effective.     

Local heating of molecular motors using single carbon nanotubes

Temperature globally affects all chemical processes and biomolecules in living cells. Elevating the temperature of an entire cell accelerates so many biomolecular reactions simultaneously that it is difficult to distinguish the various mechanisms involved. The ability to localize temperature changes to the nanometer range within a cell could provide a powerful new tool for regulating biomolecular activity at the level of individual molecules. The search for a nanoheater for biological research has prompted experiments with carbon nanotubes (CNTs), which have the highest conductivity of any known material. The adsorption of skeletal muscle myosin molecules along the length of single multi-walled CNTs (~10 μm) has allowed researchers to observe the ATP-driven sliding of fluorescently labeled actin filaments. In one study, red-laser irradiation focused on one end of a myosin-coated CNT was used to heat myosin motors locally without directly heating the surrounding water; this laser irradiation instantly accelerated the actin-filament sliding speeds from ~6 to ~12 μm/s in a reversible manner, indicating a local, real-time heating of myosin motors by approximately Δ12 K. Calculation of heat transfer using the finite element method, based on the estimated temperature along a single CNT with a diameter of 170 nm, indicated a high thermal conductivity of ~1540 Wm^?1K^?1 in solution, consistent with values measured in vacuum in earlier studies. Temperature distribution indicated by half-decrease distances was ~3660 nm along the length of the CNT and ~250 nm perpendicular to the length. These results suggest that single-CNT-based heating at the nanometer- or micrometer-range could be used to regulate various biomolecules in many areas of biological, physical, and chemical research.     

A simple method for the measurement of metabolic heat production rates during solid‐state fermentations using β‐carotene production with Blakeslea trispora as a model system

Solid‐state fermentation (SSF) technology has been rapidly developed for the past 10 years as a production platform for secondary metabolites, biofuels, food, and pharmaceuticals. Yet, the main drawback of SSF is the local temperature rise of up to 20 K, which potentially reduces the strain activity and inactivates heat sensible products. Due to the low heat capacity and thermal conductivity of mixtures of air with plant material, in comparison to aqueous suspensions in submerged fermentations, heat from metabolic processes is less efficiently dissipated. The exact knowledge of the metabolic heat generation during SSF processes is crucial to guide strategies against overheating. In this work, a simple method using a cost‐efficient multichannel instrument is proposed, which allows the determination of heat generation during SSF processes. This method was successfully tested and validated with Blakeslea trispora producing β‐carotene during growth on barley. Additionally, the consequences of the generated metabolic heat during SSF on temperature rise and water evaporation were discussed. Finally, changes in growth and product concentration could also be detected by the heat signal, implying the potential as a timesaving screening method.     

Thermal coefficients of the methyl groups within ubiquitin

Physiological processes such as protein folding and molecular recognition are intricately linked to their dynamic signature, which is reflected in their thermal coefficient. In addition, the local conformational entropy is directly related to the degrees of freedom, which each residue possesses within its conformational space. Therefore, the temperature dependence of the local conformational entropy may provide insight into understanding how local dynamics may affect the stability of proteins. Here, we analyze the temperature dependence of internal methyl group dynamics derived from the cross-correlated relaxation between dipolar couplings of two CH bonds within ubiquitin. Spanning a temperature range from 275 to 308 K, internal methyl group dynamics tend to increase with increasing temperature, which translates to a general increase in local conformational entropy. With this data measured over multiple temperatures, the thermal coefficient of the methyl group order parameter, the characteristic thermal coefficient, and the local heat capacity were obtained. By analyzing the distribution of methyl group thermal coefficients within ubiquitin, we found that the N-terminal region has relatively high thermostability. These results indicate that methyl groups contribute quite appreciably to the total heat capacity of ubiquitin through the regulation of local conformational entropy.     

Bergmann's Rule rules body size in an ectotherm: heat conservation in a lizard along a 2200‐metre elevational gradient

Bergmann's Rule predicts larger body sizes in colder habitats, increasing organisms' ability to conserve heat. Originally formulated for endotherms, it is controversial whether Bergmann's Rule may be applicable to ectotherms, given that larger ectotherms show diminished capacity for heating up. We predict that Bergmann's Rule will be applicable to ectotherms when the benefits of a higher conservation of heat due to a larger body size overcompensate for decreased capacity to heating up. We test this hypothesis in the lizard Psammodromus algirus, which shows increased body size with elevation in Sierra Nevada (SE Spain). We measured heating and cooling rates of lizards from different elevations (from 300 to 2500 m above sea level) under controlled conditions. We found no significant differences in the heating rate along an elevational gradient. However, the cooling rate diminished with elevation and body size: highland lizards, with larger masses, have a higher thermal inertia for cooling, which allows them to maintain heat for more time and keep a high body temperature despite the lower thermal availability. Consequently, the net gaining of heat increased with elevation and body size. This study highlights that the heat conservation mechanism for explaining Bergmann's Rule works and is applicable to ectotherms, depending on the thermal benefits and costs associated with larger body sizes.     

Characteristics of methotrexate polyglutamate formation in cultured hepatic cells

Upon exposure of primary monolayer cultures of hepatocytes and H35 hepatoma cells, methptrexate (MTX) is taken up by carrier-mediated mechanisms and converted to γ-glutamyl derivatives with one to four residues being added. Under conditions that result in 90% or greater conversion, the primary metabolite in both cell types is MTX with three additional glutamates (4-NH₂-10-CH₃PteGlu₄). When the time-dependent synthesis of MTX polyglutamates (4-NH₂-10-CH₃PteGlu₂ and higher) at extracellular concentrations of 10 and 100 μm methotrexate is measured, both cell types exhibit linear synthesis for 4 to 6 hr, at which time an apparent steady state intracellular concentration of approximately 40 μm is reached. The concentration of MTX polyglutamate synthesized is not due a restriction in MTX since the hepatocytes and H35 cells accumulated 400 and 138 μm intracellular methotrexate, respectively, after 24 h in the presence of 100 μm extracellular MTX. Examination of MTX polyglutamate formation following a 24-h incubation showed concentration dependence with respect to intra- and extracellular MTX. Saturation was reached at a medium concentration of approximately 2 μm with both cell types which corresponded to 10 to 12 μm intracellular MTX. Placement of cells at steady state in medium lacking MTX results in the rapid equilibration of all free intracellular MTX with the medium. The MTX polyglutamates leave the cell by a slow loss of intact polyglutamates and also by intracellular cleavage to MTX followed by efflux. The longer-chain-length γ-glutamyl derivatives (Glu_4–5) are more avidly retained by the cells than the shorter ones (Glu_2–3).     

生物热应变成像技术的研究进展

热应变成像(thermal strain imaging,TSI)是一种利用超声回波时移的温度相关性进行成像的超声应用.它既具有超声安全、无创和实时成像的优点,又能够显示与其他超声成像方式不同的组织特征,具有良好的应用前景.热应变成像目前在生物医学领域主要应用于组织表征和温度监测两个方面.本综述介绍了热应变成像的基本原理,讨论了适用于临床的主要能量源,并通过回顾近几年热应变成像的研究成果和分析目前面临的局限与挑战,对热应变成像技术的发展进行了探讨和展望.     

Intracellular pH of Thermoplasma acidophila.

Thermoplasma acidophila, a mycoplasma-like organism, was grown at 56 degrees C and pH 2. The intracellular pH of this organism is close to neutral as measured by the distribution of a radioactive weak organic acid, 5,5-dimethyl-2,4-oxazolidinedione, across the plasma membrane. The cell can maintain the pH gradient when subjected to heat or to metabolic inhibitors. Our experiments seem to indicate that a major portion of the pH gradient is not maintained by active processes, but rather by a Donnan potential across the cell membrane.     

Total internal reflection fluorescence microscopy for single-molecule imaging in living cells

Marvelous background rejection in total internal reflection fluorescence microscopy (TIR-FM) has made it possible to visualize single-fluorophores in living cells. Cell signaling proteins including peptide hormones, membrane receptors, small G proteins, cytoplasmic kinases as well as small signaling compounds have been conjugated with single chemical fluorophore or tagged with green fluorescent proteins and visualized in living cells. In this review, the reasons why single-molecule analysis is essential for studies of intracellular protein systems such as cell signaling system are discussed, the instrumentation of TIR-FM for single-molecule imaging in living cells is explained, and how single molecule visualization has been used in cell biology is illustrated by way of two examples: signaling of epidermal growth factor in mammalian cells and chemotaxis of Dictyostelium amoeba along a cAMP gradient. Single-molecule analysis is an ideal method to quantify the parameters of reaction dynamics and kinetics of unitary processes within intracellular protein systems. Knowledge of these parameters is crucial for the understanding of the molecular mechanisms underlying intracellular events, thus single-molecule imaging in living cells will be one of the major technologies in cellular nanobiology.     

Does forest fragmentation cause an increase in forest temperature?

Forest fragmentation is considered by many to be a primary cause of the current biodiversity crisis. The underlying mechanisms are poorly known, but a potentially important one is associated with altered thermal conditions within the remaining forest patches, especially at forest edges. Yet, large uncertainty remains about the effect of fragmentation on forest temperature, as it is unclear whether temperature decreases from forest edge to forest interior, and whether this local gradient scales up to an effect of fragmentation (landscape attribute) on temperature. We calculated the effect size (correlation coefficient) of distance from forest edge on air temperature, and tested for differences among forest types surrounded by different matrices using meta-analysis techniques. We found a negative edge-interior temperature gradient, but correlation coefficients were highly variable, and significant only for temperate and tropical forests surrounded by a highly contrasting open matrix. Nevertheless, it is unclear if these local-scale changes in temperature can be scaled up to an effect of fragmentation on temperature. Although it may be valid when considering “fragmentation” as forest loss only, the landscape-scale inference is not so clear when we consider the second aspect of fragmentation, where a given amount of forest is divided into a large number of small patches (fragmentation per se). Therefore, care is needed when assuming that fragmentation changes forest temperature, as thermal changes at forest edges depend on forest type and matrix composition, and it is still uncertain if this local gradient can be scaled up to the landscape.     

Heat transfer in dust structures in an RF discharge plasma

Results are presented from experimental studies of heat transfer in liquid-like plasma-dust structures. The experiments were performed with aluminum oxide grains ～3–5 μm in size in an RF discharge plasma. The heat capacity of the dust grains in plasma is measured. The thermal conductivity and thermal diffusivity of liquid-like plasma-dust structures are deduced under the assumption that the observed temperature gradients and the propagation of a thermal perturbation in a dusty plasma are related to heat conduction within the dust component. The measured temperature dependences of the thermal conductivity and thermal diffusivity are in qualitative agreement with the results of numerical simulations performed in the model of a simple single-atom liquid. It is shown that quantitative discrepancy between the experimental and numerical results is related to the energy loss of dust grains in their collisions with the neutral particles of the ambient gas.     

Intracellular pH of Thermoplasma acidophila

Thermoplasma acidophila, a mycoplasma-like organism, was grown at 56 °C and pH 2. The intracellular pH of this organism is close to neutral as measured by the distribution of a radioactive weak organic acid, 5,5-dimethyl-2,4-oxazolidinedione, across the plasma membrane. The cell can maintain the pH gradient when subjected to heat or to metabolic inhibitors. Our experiments seem to indicate that a major portion of the pH gradient is not maintained by active processes, but rather by a Donnan potential across the cell membrane.     

Entropy flow in cyclic biological processes

Biological processes are quantized, molecular, and often cyclical. Taken altogether, they comprise a heat engine working between an upper thermal reservoir at 6000 K and a lower thermal reservoir at 300 K. For optimum efficiency, such a heat engine would give out as much entropy at 300 K as it takes in at 6000 K. Yet the individual processes comprising this heat engine are quantized and molecular. What does entropy mean for them? Can we trace the entropy flow in the separate molecular cycles that are all we have when we look carefully at the overall heat engine? I think we can, and in doing so will learn why so many biological units are 5–10 μm in size.     

Dual phase lag bio-heat transfer during cryosurgery of lung cancer: Comparison of three heat transfer models

The Pennes bio-heat model is based on Fourier's law of heat conduction, which assumed that a thermal signal propagate with infinite speed. This gives contradiction in physical situation. Also, the hyperbolic bio-heat model considers the micro scale response in time, but it does not explain the micro scale response in space. Therefore, to consider the thermal behaviour which is not captured by the Fourier's law and to take into account the microstructural effect in space, a dual phase lag (DPL) bio-heat conduction model would be advantageous. In this paper, a two dimensional DPL model is proposed to study the phase change heat transfer process during cryosurgery of lung cancer. The governing equations are solved numerically by enthalpy based finite difference method. The non-ideal behaviour of tissue and heat source terms, metabolism and blood perfusion are also considered. This study is made to examine the effects of phase lags in heat flux and temperature gradient on interface positions and temperature distribution during freezing process. A comparative study of DPL, parabolic and hyperbolic conduction models is thoroughly investigated. It is found that the phase lags of temperature gradient and heat flux have significant effect on interface positions and temperature distribution.     

Bacterial Thermotaxis by Speed Modulation

Naturally occurring gradients often extend over relatively long distances such that their steepness is too small for bacteria to detect. We studied the bacterial behavior in such thermal gradients. We find that bacteria migrate along shallow thermal gradients due to a change in their swimming speed resulting from the effect of temperature on the intracellular pH, which also depends on the chemical environment. When nutrients are scarce in the environment the bacteria''s intracellular pH decreases with temperature. As a result, the swimming speed of the bacteria decreases with temperature, which causes them to slowly drift toward the warm end of the thermal gradient. However, when serine is added to the medium at concentrations >300 μM, the intracellular pH increases causing the swimming speed to increase continuously with temperature, and the bacteria to drift toward the cold end of the temperature gradient. This directional migration is not a result of bacterial thermotaxis in the classical sense, because the steepness of the gradients applied is below the sensing threshold of bacteria. Nevertheless, our results show that the directional switch requires the presence of the bacterial sensing receptors. This seems to be due to the involvement of the receptors in regulating the intracellular pH.     

Thermal imaging of receptor-activated heat production in single cells.

Changes in enthalpy (i.e., heat content) occur during the diverse intracellular chemical and biophysical interactions that take place in the life cycle of biological cells. Such changes have previously been measured for cell suspensions or cell-free biochemical extracts by using microcalorimetry, thermocouples, or pyroelectric films, all of which afford minimal spatial or temporal resolution. Here we present a novel thermal imaging method that combines both diffraction-limited spatial (approximately 300 nm) and sampling-rate-limited time resolution, using the temperature-dependent phosphorescence intensity of the rare earth chelate Eu-TTA (europium (III) thenoyltrifluoro-acetonate). With this thermosensitive dye, we imaged intracellular heat waves evoked in Chinese hamster ovary cells after activation of the metabotropic m1-muscarinic receptor. Fast application of acetylcholine onto the cells evoked a biphasic heat wave that was blocked by atropine, and after a brief delay was followed by a calcium wave. Atropine applied by itself produced a monophasic heat wave in the cells, suggesting that its interactions with the receptor activate some intracellular metabolic pathways. The thermal imaging technique introduced here should provide new insights into cellular functions by resolving the location, kinetics, and quantity of intracellular heat production.     

Thermal chip fabrication with arrays of sensors and heaters for micro-scale impingement cooling heat transfer analysis and measurements

The design and fabrication for a thermal chip with an array of temperature sensors and heaters for study of micro-jet impingement cooling heat transfer process are presented. This thermal chip can minimize the heat loss from the system to the ambient and provide a uniform heat flux along the wall, thus local heat transfer processes along the wall can be measured and obtained. The fabrication procedure presented can reach a chip yield of 100%, and every one of the sensors and heaters on the chip is in good condition. In addition, micro-jet impingement cooling experiments are performed to obtain the micro-scale local heat transfer Nusselt number along the wall. Flow visualization for the micro-impinging jet is also made. The experimental results indicate that both the micro-scale impinging jet flow structure and the heat transfer process along the wall is significantly different from the case of large-scale jet impingement cooling process.