Water proton magnetic resonance studies of normal and sickle erythrocytes Temperature and volume dependence

The temperature and cell volume dependence of the NMR water proton linewidth, spin-lattice, and spin-spin relaxation times have been studied for normal and sickle erythrocytes as well as hemoglobin A and hemoglobin S solutions. Upon deoxygenation, the spin-spin relaxation time (T₂) decreases by a factor of 2 for sickle cells and hemoglobin S solutions but remains relatively constant for normal cells and hemoglobin A solutions. The spin-lattice relaxation time (T₁) shows no significant change upon dexygenation for normal or sickle packed red cells. Studies of the change in the NMR linewidth, T₁ and T₂ as the cell hydration is changed indicate that these parameters only slightly by a 10–20% cell dehydration. This result suggests that the reported 10% cell dehydration observed with sickling is not important in the altered NMR properties. Low temperature studies of the linewidth and T₁ for oxy and deoxy hemoglobin A and hemoglobin S solutions suggest that the “bound” water possesses similar properties for all four species. The low temperature linewidth ranges from about 250 Hz at ?15°C to 500 Hz at ?36°C and analysis of the NMR curves yield hydration values near 0.4 g water/g hemoglobin for all four species. The low temperature T₁ data go through a minimum at ?35°C for measurements at 44.4 MHz and ?50°C for measurements at 17.1 MHz and are similar for oxy and deoxy hemoglobin A and hemoglobin S. These similarities in the low temperature NMR data for oxy and deoxy hemoglobin A and hemoglobin S suggest a hydrophobically driven sickling mechanism. The room temperature and low temperature relaxation time data for normal and sickle cells are interpreted in terms of a three-state model for intracellular water. In the context of this model the relaxation time data imply that type III, or irratationally bound water, is altered during the sickling process.     

Factors affecting molecular packing in mixed lipid monolayers and bilayers

The nature of the molecular interactions and the factors determining molecular packing in mixed phospholipid/glyceride monolayers and bilayers were investigated by monolayer and nuclear magnetic resonance (NMR) techniques. Force-area curves were obtained at various temperatures for monolayers, at the air-water interface, of synthetic lecithins and a phosphatidyl ethanolamine mixed with di- and triglycerides in different molar ratios. The linewidths of peaks in the high resolution NMR spectra of lecithin/glyceride co-dispersions in excess water at different temperatures were used to obtain information about molecular mobilities.It was found that the molecular packing in mixed lipid monolayers and bilayers is determined by the following factors: (1) Whether lipid chains are above or below their melting point (T_C). (2) The difference between experimental temperature and T_C: the larger the difference, the smaller the effect of one component on the other. (3) The degree of similarity of the chains of the components; this influences the degree of cooperativity of chain motions and the degree of mixing of the components. (4) The nature, orientation, mutual interaction and degree of hydration of the polar groups.It is shown that mean molecular area does not always reflect the state of chain motions in lipid films, because of heterogeneity of motion and structure along the molecules. Cooperativity of motion may reduce steric requirements; other effects which are of particular importance for lecithins are interactions of zwitterions, and the influence of polar group hydration.     

Occurrence of heat-dissociable ribosomal RNA in insects: the presence of three polynucleotide chains in 26 S RNA from cultured Aedes aegypti cells

When RNA extracted from a mixture of cultured mosquito (Aedes aegypti) and hamster (BHK) cells is heated at 60 °C for five minutes the 26 S mosquito RNA but not the 28 S BHK RNA is converted to 18 S products. These products are not separable from each other or from pre-existent 18 S RNA on 2.4% acrylamide gels and have molecular weights near 0.7 × 10⁶. The large ribosomal RNA from insects belonging to ten different orders shows a similar conversion, although this property is absent in two species of aphid.A. aegypti 26 S RNA dissociates over a narrow temperature range. The reaction equilibrium favours dissociation and is dependent on ionic strength, showing a 6 deg. C change in T_m′ (the temperature of 50% dissociation) with tenfold change in salt concentration. Although the T_m of 26 S RNA from Drosophila melanogaster and A. aegypti is markedly different, reflecting the difference in base composition, the T_m′ of the two RNA species was virtually the same.High molecular weight ribosomal RNA from Escherichia coli, BHK cells and A. aegypti cells was terminally labelled with [³H]isonicotinic acid hydrazide. The specific activities of the large RNA species show the presence of one, two and three polynucleotide chains in 23 S, 28 S and 26 S RNA, respectively. A. aegypti 26 S RNA contains a small, heat-dissociable “IRNA” similar in relative amount and mobility to that found in BHK cells.     

Studies on lactate dehydrogenase activity in the shrimp Palaemon serratus. Part 1: Characterization of the enzymes from the caudal muscle of Palaemon serratus

Palaemon serratus muscle lactate dehydrogenase (LDH) has been partially purified. The electrophoretic pattern of the LDH enzymes indicates that at least two molecular structures are present. The mean molecular weight is 130,000. The substrate and coenzyme dependence of the LDH system show non-Michaelian kinetics. This can be interpreted as being due to the presence of two binding sites in the enzyme which show negative effects. The behaviour of the two molecular species of LDH appears to be nearly identical in this respect. The study of the effect of temperature on the kinetic parameters of the LDH system shows the existence of a thermal dependence of K_m values. This may be related to developmental or environmental changes in the animal.     

Low temperature study of myoglobin-ligand rebinding kinetics with Mössbauer spectroscopy

We have studied the recombination kinetics of carboxymyoglobin (after photodissociation of the CO ligand) by Mössbauer spectroscopy for temperatures in the range 4.2 – 60 K. The observed kinetics display non-exponential behaviour which was monitored over periods of a few days. It is shown that the time dependence of the kinetics can be reduced to a single universal function of the temperature-dependent variable (t/τ _1/2(T)) ^β(T) . The half-decay time τ _1/2(T) and the scaling parameter β(T) are analysed for the presence of tunneling effects. The non-Arrhenius temperature dependence of the half-decay time below 60 K is interpreted as activated tunneling in models with an Eckart barrier or a fluctuating barrier.     

Thermal ecology of five Cnemidophorus species (Squamata: Teiidae) in east coast of Brazil

In this study we compared the body temperature of 16 populations belonging to five species of the genus Cnemidophorus from restinga habitats along the eastern coast of Brazil in order to evaluate the importance of how some environmental factors affect lizard body temperatures. Cloacal body temperatures (T_b) were taken immediately after capture with a quick-reading thermometer (Schultheis). Substrate temperatures (T_s) and air temperatures (T_a; approximately 1 cm above the substrate) were taken as close as possible to the point when each lizard was initially sighted. Most of the mean body temperatures in activity of the different populations and species of Cnemidophorus along the coast of Brazil ranged from 36.5 to 39.3 °C, except for Cnemidophorus lacertoides (T_b=35.2 °C) in the restinga of Joaquina, SC and for Cnemidophorus ocellifer (T_b=34.8 °C ) in the restinga of Praia do Porto, SE. Some studies show that the body temperature of lizards is more related to phylogenetic than ecological factors, suggesting that species of the same genus tend to have similar body temperatures even occurring in different types of environments. In general, regardless of the locality and latitude along the eastern coast of Brazil, the different species of lizards of the genus Cnemidophorus and their respective populations have similar body temperatures in activity and the apparent differences result from the influence of the local thermal environment of each restinga.     

Influence of α-branched fatty acid chains on the thermotropic behaviours of 1-O-acyl-2-O-hexadecyl-glycerophosphocholines

Phosphatidylcholines containing a branched-chain fatty acid in 1-position at the glycerol backbone were synthesized and characterized by differential scanning calorimetry; We find that their thermotropic phase behaviour depends on the length of the branches. The results show that, depending on the side-chain length, the gel-to-liquid-crystalline phase transition temperatures (T_m) pass through a minimum. The systematic change of the T_m-values is connected with a modified temperature dependence of the apparent molar heat capacities.     

Is locomotor performance optimised at preferred body temperature? A study of Liolaemus pictus argentinus from northern Patagonia, Argentina

We studied the relationship between locomotor performance and temperature in Liolaemus pictus argentinus, from the Andean-Patagonian forest, Argentina. We determined the running speed in long and sprint runs at four different body temperatures, the panting threshold, and minimum critical temperature. The results are discussed in relation to body temperature in the field and thermal preference in the laboratory (T_pref). L. p. argentinus achieved higher speed in sprint runs than in long runs at all temperatures. In order to know if pregnancy constrains performance in this viviparous species, the differences between pregnant females and the other adults were analysed. Pregnant females were at a disadvantage when running long distances, but in sprint runs they were able to run as efficiently as the rest of the individuals, suggesting that they mainly use sprint runs and this may explain their conspicuous more-withdrawn behaviour. In long runs, the performance optimal temperature for L. p. argentinus (T_o=30.7 °C) was below the 25th percentile for all body temperatures selected in the laboratory (set-point range of T_pref=34.6-37.9 °C), but similar to the mean field body temperature (32.1 °C). However, in sprint runs the T_o (36.3 °C) was within the set-point range of T_pref. The mean panting threshold (42.8 °C) and the mean minimum critical temperature (6.9 °C) were similar to those of other liolaemids. The results are evidence that L. p. argentinus is well-adapted to the temperatures available in their environment and that the species has a T_pref that allows the achievement of maximal locomotor performance in the most frequently used and probably the most important run type, the sprint run.     

Thermoregulation in the cicada Platypedia putnami variety lutea (Homoptera: Tibicinidae) with a test of a crepitation hypothesis

• 1.Measurements of body temperature (T_b) in the field demonstrate that Platypedia putnami var. lutea Davis regulates T_b through behavioral mechanisms.
• 2.Thermal responses (minimum flight temperature 17.3°C, maximum voluntary tolerance-temperature 32.5°C, and heat torpor temperature 44.4°C) of P. putnami var. lutea are related to the altitude of their habitat.
• 3.Water loss rates increase with ambient temperature (T_a). Water loss rates are not significantly different at the extremes of the active T_b range but increase significantly when exposed to elevated T_a.
• 4.Acoustic activity was restricted at 6.7°C T_b range. This is similar to the lower end of the T_b range for singing measured in cicada species that produce sound with a timbal mechanism.
• 5.The use of the wing musculature to produce acoustic signals in P. putnami var. lutea does not increase the T_b range over which the species can call compared to timbal calls produced by other cicada species.

     

A cold-adapted reptile becomes a more effective thermoregulator in a thermally challenging environment

Thermoregulation is of great importance for the survival and fitness of ectotherms as physiological functions are optimized within a narrow range of body temperature (T _b). The precision with which reptiles thermoregulate has been proposed to be related to the thermal quality of their environments. Although a number of studies have looked at the effect of thermal constraints imposed by diel, seasonal and altitudinal variation on thermoregulatory strategies, few have addressed this question in a laboratory setting. We conducted a laboratory experiment to test whether tuatara, Sphenodon punctatus (order Rhynchocephalia), a cold-adapted reptile endemic to New Zealand, modify their thermoregulatory behaviour in response to different thermal environments. We provided tuatara with three thermal treatments: high-quality habitat [preferred T _b (T _sel) could be reached for 8 h/day], medium-quality habitat (T _sel available for 5 h/day) and low-quality habitat (T _sel available for 3 h/day). All groups maintained body mass, but tuatara in the low-quality habitat thermoregulated more accurately and tended to maintain higher T _bs than tuatara in the high-quality habitat. This study thus provides experimental evidence that reptiles are capable of adjusting their thermoregulatory behaviour in response to different thermal constraints. This result also has implications for the conservation of tuatara. A proposed translocation from their current habitat to a higher latitudinal range within New Zealand (similar to the shift from our 8 h/day to our 5 h/day regime) is unlikely to induce thermoconformity; rather, tuatara will probably engage in more effective thermoregulatory behaviour.     

The amorphous state of spray-dried maltodextrin: sub-sub-Tg enthalpy relaxation and impact of temperature and water annealing

The annealing behaviour of a spray-dried maltodextrin was investigated by differential scanning calorimetry. Special attention was paid to the effect of temperature and humidity on the annealing process. Comparison was also made with the glassy state of the same compound prepared by various cooling processes. The presence of a very pronounced sub-T_g peak upon ageing reveals the specificities of the glass and the complexity of the relaxation spectrum of the spray-dried material. This peak seems actually to correspond to a partial ergodicity recovery that may be attributed to onset of molecular mobility occurring below T_g. The position of the sub-T_g peak with regard to the conventional T_g was systematically studied. It clearly showed the difference between the effect of temperature and water plasticization on the relaxations occurring in the glassy state of materials prepared by spray-drying.     

Direct observation of the properties of cholesterol in membranes by deuterium NMR

The properties of cholesterol in bilayers of egg phosphatidylcholine (PC) were investigated directly by means of ²H-NMR of specifically-deuterated species (C3, C7, C26, C27). Quadrupole splittings were a measure of molecular ordering, and relaxation times T₁ and T_2e were indicators of rates of motion. The importance of the use of echoes for spectral acquisition is emphasised, particularly to obtain accurate values of the quadrupole splitting. In the case of overlapping powder patterns from two labelled positions, the use of the absolute value mode of spectral presentation is shown to yield reasonable estimates of the individual quadrupole splittings. Spectral properties were monitored as a function of cholesterol concentration and temperature. Increasing cholesterol concentration led to a high degree of ordering for the rigid ring system of cholesterol, approaching a molecular order parameter of 0.8 at 50 mol% cholesterol. The isopropyl methyl groups were in all cases less ordered anmore mobile than the ring system, but responded in a similar fashion to variable cholesterol concentration and temperature. The observation of a minimum in the temperature dependence of T₁ for cholesterol-7,7-d₂ led to a direct estimate of its correlation time for molecular motion, 3.5 × 10^?9 s rad^?1. This indicates that the overall rate of motion of cholesterol is considerably slower than that of the lipids in which it is located. The short T_2e values suggest that the motional spectrum of cholesterol is rich in low frequencies. The parallel temperature and cholesterol dependences of quadrupole splittings for different positions on the rigid ring system of cholesterol indicate that the position of the axis of motional averaging of the molecule is not changing, and is the same as that determined in an earlier study. It is emphasised that the steep temperature dependence and small quadrupole splittings for the chain isopropyl methyl groups of cholesterol do not necessarily indicate a high degree of disorder, but may be due to their axes of motional averaging lying at angles close to 54° with respect to the director of the ordered lipids.     

Body temperature patterns in two syntopic elephant shrew species during winter

We measured body temperature (T_b) in free-ranging individuals of two species of elephant shrews, namely western rock elephant shrews (Elephantulus rupestris) and Cape rock elephant shrews (E. edwardii), during winter in a winter-rainfall region of western South Africa. These syntopic species have similar ecologies and morphologies and thus potential for large overlaps in diet and habitat use. Unexpectedly, they displayed different T_b patterns. Western rock elephant shrews were heterothermic, with all individuals decreasing T_b below 30 °C on at least 34% of nights. The level of heterothermy expressed was similar to other species traditionally defined as daily heterotherms and was inversely related to T_a, as is commonly seen in small heterothermic endotherms. In contrast, Cape rock elephant shrews rarely allowed their T_b to decrease below 30 °C. The level of heterothermy was similar to species traditionally defined as homeotherms and there was no relationship between the level of heterothermy expressed and T_a. In both species, the minimum daily T_b was recorded almost exclusively at night, often shortly before sunrise, although in some individuals minimum T_b occasionally occurred during the day. The interspecific variation in T_b patterns among Elephantulus species recorded to date reiterates the importance of ecological determinants of heterothermy that interact with factors such as body mass and phylogeny.     

Thermal biology of Liolaemus lizards from the high Andes: Being efficient despite adversity

We studied the efficiency of thermoregulation in four high elevation Liolaemus species in the Andes of Salta, Argentina; Liolaemus irregularis, Liolaemus multicolor, Liolaemus albiceps and Liolaemus yanalcu. One of the species, L. irregularis, shows a broad distribution being in allopatry in some localities and in sympatry with L. albiceps, L. multicolor and L. yanalcu at different sites. Together with this variation in assemblages, the degree of phylogenetic relatedness is different with L. irregularis being most closely related to L. albiceps than to the other two species (L. multicolor and L. yanalcu). We measured body (T_b), microenvironmental (T_a, T_s), and operative temperatures (T_e) in the field, and preferred body temperature (T_pref) in laboratory for each one of the species of assemblages. Three out of the four species showed a high thermoregulatory efficiency except for L. yanalcu, a moderate thermoregulator. The species studied here show high T_b in the field compared to most of the recorded Liolaemus species. However, the T_pref values were similar to other Liolaemus species. No evidence of thermal niche segregation between species in sympatry was observed. Our results suggest that the species studied here, despite living at high elevation and harsh climatic conditions are able to behaviorally or physiologically thermoregulate to achieve T_bs close to their T_pref, probably because of low predation risk and perhaps low levels of competition.     

Absence of heterothermy in the European red squirrel (Sciurus vulgaris)

The European red squirrel Sciurus vulgaris inhabits areas which undergo profound seasonal declines in food availability and ambient temperature. We measured the body temperature (T_b) of free-ranging S. vulgaris over the course of one year to examine its thermoregulatory strategies and found no evidence of heterothermy, with T_b never dropping below 36.7 °C. A lower average T_b and a reduced active phase are likely to have resulted in some energetic savings, sufficient for survival during the particularly mild winter with unhindered access to food stores. We cannot exclude that a different T_b pattern may be seen in energetically more demanding years, but we can confirm that heterothermy is not an obligatory behaviour in this species to counter energetic bottlenecks. Either S. vulgaris is indeed a strict homeotherm, or the need for torpor is flexibly adjusted.     

Spin labeling EPR studies of the properties of oxidized phospholipid-containing lipid vesicles

This study aims at characterizing the structure and some properties of phospholipid multi-lamellar vesicles (MLVs) containing the oxidized species γ-palmitoyl-β-(9-hydroperoxy-10,12-octadecanedienoyl)-lecithin (HPPLPC), γ-palmitoyl-β-(9-hydroxy-10,12-octadecanedienoyl)-lecithin (HOPLPC), γ-palmitoyl-β-glutaroyl-lecithin (GlPPC) and γ-palmitoyl-β-azelaoyl-lecithin (AzPPC). Sepharose 4B gel-chromatography was used to ensure and check that only MLVs are used in EPR measurements. Gel-solid to gel-liquid transition temperature (T_m), lateral phase separation, fluidity gradient and polarity profile were studied by use of EPR spectroscopy of enclosed n-doxylstearoyl lecithin spin labels. Contrarily to conjugate dienes and normal phospholipids, pure carboxyacyl species yielded aqueous suspensions showing gel-chromatography elution profile resembling that of lysolecithin micelles. Conjugate dienes/DPPC MLVs showed lateral phase separation at room temperature and T_m value lower than pure DPPC MLVs. Pure conjugate dienes MLVs resembled more PLPC MLVs and displayed free miscibility with PLPC in mixed MLVs. Pure HPPLPC MLV bilayer appeared to be slightly more rigid, while that of HOPLPC and the polarity profile of MLVs made of the pure conjugate dienes species were similar to those of normal PLPC. It is concluded that carboxyacyl lecithins in MLVs tend to disrupt vesicle structure, while conjugated dienes lecithins are more able to affect some physical properties of the bilayer, and that DPPC in MLVs enhances these effects while PLPC shows a better compatibility with the lipoperoxides.     

Interspecific variation in thermoregulation among three sympatric bats inhabiting a hot, semi-arid environment

Bats in hot roosts experience some of the most thermally challenging environments of any endotherms, but little is known about how heat tolerance and evaporative cooling capacity vary among species. We investigated thermoregulation in three sympatric species (Nycteris thebaica, Taphozous mauritianus and Sauromys petrophilus) in a hot, semi-arid environment by measuring body temperature (T _b), metabolic rate and evaporative water loss (EWL) at air temperatures (T _a) of 10?C42?°C. S. petrophilus was highly heterothermic with no clear thermoneutral zone, and exhibited rapid increases in EWL at high T _a to a maximum of 23.7?±?7.4?mg?g^?1?h^?1 at T _a????42?°C, with a concomitant maximum T _b of 43.7?±?1.0?°C. T. mauritianus remained largely normothermic at T _as below thermoneutrality and increased EWL to 14.7?±?1.3?mg?g^?1?h^?1 at T _a????42?°C, with a maximum T _b of 42.9?±?1.6?°C. In N. thebaica, EWL began increasing at lower T _a than in either of the other species and reached a maximum of 18.6?±?2.1?mg?g^?1?h^?1 at T _a?=?39.4?°C, with comparatively high maximum T _b values of 45.0?±?0.9?°C. Under the conditions of our study, N. thebaica was considerably less heat tolerant than the other two species. Among seven species of bats for which data on T _b as well as roost temperatures in comparison to outside T _a are available, we found limited evidence for a correlation between overall heat tolerance and the extent to which roosts are buffered from high T _a.     

Acclimation of photosynthetic temperature optima of temperate and boreal tree species in response to experimental forest warming

Rising temperatures caused by climate change could negatively alter plant ecosystems if temperatures exceed optimal temperatures for carbon gain. Such changes may threaten temperature‐sensitive species, causing local extinctions and range migrations. This study examined the optimal temperature of net photosynthesis (T_opt) of two boreal and four temperate deciduous tree species grown in the field in northern Minnesota, United States under two contrasting temperature regimes. We hypothesized that T_opt would be higher in temperate than co‐occurring boreal species, with temperate species exhibiting greater plasticity in T_opt, resulting in better acclimation to elevated temperatures. The chamberless experiment, located at two sites in both open and understory conditions, continuously warmed plants and soils during three growing seasons. Results show a modest, but significant shift in T_opt of 1.1 ± 0.21 °C on average for plants subjected to a mean 2.9 ± 0.01 °C warming during midday hours in summer, and shifts with warming were unrelated to species native ranges. The 1.1 °C shift in T_opt with 2.9 °C warming might be interpreted as suggesting limited capacity to shift temperature response functions to better match changes in temperature. However, T_opt of warmed plants was as well‐matched with prior midday temperatures as T_opt of plants in the ambient treatment, and T_opt in both treatments was at a level where realized photosynthesis was within 90–95% of maximum. These results suggest that seedlings of all species were close to optimizing photosynthetic temperature responses, and equally so in both temperature treatments. Our study suggests that temperate and boreal species have considerable capacity to match their photosynthetic temperature response functions to prevailing growing season temperatures that occur today and to those that will likely occur in the coming decades under climate change.     

Evaluation of the binding constants,bound chemical shifts,and equilibrium stoichiometry for the association of lanthanide shift reagents with carbohydrate derivatives

The molecular associations of tris(dipivalomethanato)europium(III) (1) and of tris(2,2-dimethyl-6,6,7,7,8,8,8,-heptafluoro-3,5-octanedionato)europium(III) (2) with six carbohydrate derivatives have been studied. The interactions of 2 in deuterochloroform solution appear to be “normal”, i.e., the plot of [sugar concentration] versus [induced chemical-shift change]^?1 is linear and can be interpreted in terms of the equilibrium binding constant (K_B) and the bound chemical shifts (°_B). Similar results are found for 1 in carbon tetrachloride solutions. However, in chloroform solutions, non-linear plots are obtained that cannot be simply interpreted. Possible reasons for this behaviour are given.     

Conductance behavior of symmetrical tetraalkylammonium halides in aqueous sucrose solutions

The conductance behavior of some tetraalkylammonium halides (R₄NX) in saturated, aqueous solutions of sucrose has been investigated, and data on the conductance of these salts in water saturated by sucrose at 50° are reported at several temperatures within the range 25 to 70°. In these homogeneous, ternary systems, plots of —log K versus 1/T show a break at the saturation temperature, where two straight lines intersect one another. Divergence of the pair of straight lines has been found to decrease with increasing chain-length of the R₄N⁺ ions, in contrast to the structural behavior of common, alkali-metal ions. The results are interpreted in terms of the hydrophobic nature of the tetraalkylammonium halides, as well as the salting-in behavior of these salts towards sucrose molecules.