Variations in the activity of fatty ACYL-CoA desaturases and electron-transport components in Tetrahymena microsomes with temperature and age of culture

1. 1.|Alterations in the fatty acid composition of microsomes were most marked in the exponential phase of both 39.5- or 15°C- grown Tetrahymena pyriformis NT-1.

2. 2.|Activities of palmitoyl-CoA and stearoyl-CoA desaturases were lower in 15°C cells than in 39.5°C cells, while the activity of oleoyl-CoA desaturase was higher in 15°C cells.

3. 3.|Activities of the terminal component of the desaturation system as well as all three desaturases (palmitoyl-CoA, stearoyl-CoA, oleoyl-CoA) were higher in the exponential phase than in the stationary phase for cells grown at both temperatures.

4. 4.|NAD(P)H-cytochrome c reductase activity and cytochrome b₅ content were reduced whereas NADH-ferricyanide reductase activity was increased in the stationary phase at both 39.5 and 15°C.

Hyperthermia-induced changes in the morphology of CHO-K1 and their refractile inclusions

1. 1.|Chinese hamster ovary cells (CHO-K1) were heated at temperatures of 42°C and above.

2. 2.|Cells were cultured in microcapillaries to eliminate handling stress, and morphological changes were observed by light microscopy.

3. 3.|Increased incidence of membrane blebbing was noted between 1 and 2 h and few cells were viable after 2 h at 43°C.

4. 4.|Morphological changes, including the appearance of potocytotic blebs, were recorded by cinemicroscopy of microcapillary cultures on a heated microscope stage.

5. 5.|Lipid-rich refractile cell inclusions changed shape before blebbing occurred.

6. 6.|Cell retraction and rearrangement of organelles seen at 1 h at 43°C are the reverse of those seen in spreading post-trypsinized cells and suggest a thermal effect on the cytoskeleton.

Thermal resistance of the ciliary activity in the gills of the fresh water mussel Anodonta anatina

1. 1.|The thermal resistance of the activity of frontal cilia in the median gills of the fresh water mussel Anodonta anatina was studied.

2. 2.|The resistance acclimation appeared in 2 days in the gills of intact animals, but not in the isolated gills kept at 4, 14 and 24°C, for between 1 to 3 days.

3. 3.|Warm-acclimation increased the ACh sensitivity of the gills of intact mussels.

4. 4.|Isolation of the gills enhanced the thermal resistance.

5. 5.|ACh, choline and tetramethylammonium enhanced the thermal resistance in the isolated gills. whereas atropine and physostigmine diminished it.

6. 6.|It is concluded that in A. anatina the control if the thermal resistance is probably neural.

Acute adjustment of thermal tolerance in vertebrate ectotherms following exposure to critical thermal maxima

1. 1.|Heat hardening in a transitory increase in heat tolerance following a sublethal exposure to lethal high temperatures.

2. 2.|Within 1–2 h of an initial exposure to the critical thermal maximum (CTM), the CTM of two species of amphibians and two species of fish had increased significantly above the initial level and then decreased to the initial level within 24 h.

3. 3.|Experiments with exposure to sub-CTM temperatures and multiple exposures to the CTM indicated that hardening requires exposure to the CTM and may be the maximum CTM attainable by the animal.

4. 4.|Diel and seasonal variation had significant effects on hardening ability.

5. 5.|Field evidence suggests that heat hardening is adaptive in that it provides an acute means of adjustment to extreme fluctuations in diurnal temperatures.

Thermal acclimation and hardening in tadpoles of the bullfrog, Rana catesbeiana

1. 1.|Critical thermal maxima (CTMax) and minima (CTMin) were measured to evalute thermal hardening in Rana catesbeiana.

2. 2.|Tadpoles show heat hardening and CTMax acclimation, and both responses are influenced by developmental stage.

3. 3.|The first evidence of cold hardening in vertebrates is reported here.

4. 4.|Heat hardening significantly reduces cold tolerance, but there is otherwise no evidence of a cross-hardening effect.

The effect of thermal stress on Ostrini00a nubilalis HBN (Lepidoptera, Pyralidae)—I. The orle of the neurosecretory cells in the survival of diapausing larvae

1. 1.|The effect of short-term exposure (up to 12 h) to upper limiting temperature (45°C) on A-type medial neurosecretory cells (MNSC) of the protocerebrum of Ostrinia nubilalis larvae was studied.

2. 2.|For evaluation of the activity of MNSC, cytological parameters were used: quantity of the neurosecretory material, size of the neurosecretory granules, shape of the nuclei and size of the nucleoli.

3. 3.|After 1 h exposure to 45°C the activity of the MNSC was increased, after 3 h it was decreased, while after 6 h exposure to the same temperature some renewed increase in the activity was observed.

4. 4.|The significance of oscillatory changes in the activity of A-type MNSC in survival of the thermal stress of O. nubilalis diapausing larvae is discussed.

Heat production of mammalian cells at different cell-cycle phases

1. 1.|Heat production of Reuber H35 rat hepatoma cells and murine C1300 neuroblastoma cells at different stages of the cell cycle were measured microcalorimetrically.

2. 2.|Reuber H35 monolayer cultures of G1-phase cells and cells in S-phase were trypsinized, reincubated in suspension culture and immediately used for microcalorimetric measurements. There was a remrkable difference in the heat evolution of H35-cells in suspension derived from a monolayer culture of G1-phase cells and that of cells in S-phase of the cell cycle. Heat production of G1-cells was relatively continuous during the experiment, in contrast to S-phase cells that showed a decrease in heat production after an initial maximum.

3. 3.|Neuroblastoma cells synchronized by mitotic shake-off and cultured in suspension progressed through their cell cycle. They showed maximal heat production shortly before and durign mitosis.

Tolerances of Nauplii of Balanus balanoides (L.) and Elminius modestus (Darwin) exposed to cyclical temperature fluctuations

1. 1.|Newly-released larvae of both E. modestus and B. balanoides show similar tolerances to steady temperature levels.

2. 2.|Both species show an ability to withstand a wider range of temperatures when subjected to cyclic thermal shocks.

3. 3.|E. modestus shows a greater tolerance to short-term thermal shocks than B. balanoides.

The effect of incubation temperatuer on oxygen consumption and organ growth in domestic-fowl embryos

1. 1.|Oxygen consumption and organ growth were measured in domestic-fowl embryos incubated at different temperatures (36, 38 and 40°C).

2. 2.|Embryonic oxygen consumption was highest at an incubation temperature of 40°C and lowest at 36°C. These differences were ascribed largely to variations in embryo size at different incubation temperatures.

3. 3.|At incubation temperatuers of 40 and 38°C, there was a plateau in oxygen consumption late in incubation, but this was not apparent at 36°C.

4. 4.|At 36°C, some tissues (e.g. eyeballs) were “spared” the repression of growth that characterized the embryo as a whole, while other tissues (e.g. stomach) incurred a much greater growth reduction. Similarly, at 40°C, stomach growth exceeded that of the embryo as a whole, while the eyeballs were largely spared the enhanced growth.

5. 5.|A simple index of tissue age revealed that, in general, there were consensual changes in tissue maturity and growth at different temperatures but that there were some disparities between growth and maturity in individual organs.

Thermohaemolysis of human erythrocytes in sucrose containing isotonic media

1. 1.|The thermohaemolysis of human erythrocytes in NaCl/sucrose isotonic media can be best accounted for in terms of the colloid-osmotic theory of haemolysis.

2. 2.|The thermohaemolysis in NaCl saline was preceded by leakage of K⁺ and cell swelling. If the inner oncotic osmoactivity was balanced with external sucrose the cells progressively shrinked losing K⁺, but the haemolysis was strongly reduced.

3. 3.|Time dependence of the shrinking of cells and one-step resealed ghosts suspended in isotonic 60 mOsm NaCl/sucrose media was studied between 50 and 58°C.

4. 4.|After a lag period for cells only, this shrinking proceeded with apparently constant rate for cells and ghosts.

5. 5.|The rate constant of shrinking for cells and ghosts obeys the Arrhenius relation, giving the value of 250 ± 15 kJ/mol for the activation energy of shrinking in both cases. This is also the case for the activation energy of the membrane ion permeability constant.

6. 6.|These results are consistent with the thermal inactivation of membrane associated protein(s) acting as a trigger for the ion permeability barrier disturbance.

7. 7.|The mid-point temperature for these membrane events was about 61°C.

Alteration of heat sensitivity by treatment with nonpermissive temperature or cycloheximide in temperature-sensitive CHO mutant tsH1 cell

1. 1.|The temperature-sensitive mutant CHO-tsH1 and wild type (CHO-SC) cells became thermal resistant when cells were treated for either 2 h at 39.5°C before heating at 43°C or 2 h with 10 μg/ml cycloheximide (CHM) before and during heating at 43°C.

2. 2.|There was a 2000-fold increase in survival after 2.5 h at 43°C by preincubation at 39.5°C in both cell types. There was also a 200- or 700-fold increase in survival after 2.5 h at 43°C by treatment with CHM in tsH1 or SC cell type respectively.

3. 3.|In contrast to the effects at 43°C, at 41.8°C these protective effects were not evident in tsH1 cells. In wild type, however, there was an 800- or 1800-fold increase in survival after 8 h at 41.8°C by preincubation at the temperature of 39.5°C or treatment with CHM, respectively.

4. 4.|Therefore, these results suggest that killing of tsH1 at low temperature hyperthermia (41.8°C) is probably due to denaturation of thermolabile leucyl-tRNA synthetase.

5. 5.|The denaturation of this enzyme may not be protected by inhibition of protein synthesis by preincubation at the nonpermissive temperature of 39.5°C or by CHM.

Effect of cycloheximide on nonpermissive temperature killing in tsH1 mutant cells

1. 1.|We investigated the mechanism of cycloheximide-induced heat protection. We proposed a hypothesis to account for the mechanism [Lee and Dewey (1986) Radiat. Res. 106, 98–110].

2. 2.|Cycloheximide protects cells from hyperthermic killing by means of protecting thermolabile proteins from denaturation.

3. 3.|For this study, we have employed temperature-sensitive mutant tsH1 which contains a thermolabile leucyl-tRNA synthetase.

4. 4.|By 15 h of incubation at the nonpermissive temperature of 39.5 or 40°C, 40 or 93% of mutant cells respectively, were killed. In contrast, wild type SC cells did not lose viability after this same incubation.

5. 5.|Although killing of tsH1 by incubation at the nonpermissive temperatures was mainly due to denaturation of a thermolabile leucyl-tRNA synthetase, cycloheximide did not protect mutant cells from killing. However, tsH1 and SC cells exhibited similar sensitivities to killing at 43°C and above. Furthermore, cycloheximide protected both cell types from hyperthermic killing.

6. 6.|There was a 200- or 700-fold increase in survival after 2.5 h at 43°C by treatment with cycloheximide in tsH1 or SC cell type, respectively. Thus, the cellular target(s) for hyperthermic killing at this temperature apparently are similar in both types of cells.

7. 7.|The data indicate that the mechanism behind cycloheximide-induced heat protection is probably not the prevention of protein denaturation.

Influence of temperature changes on the ability of gangliosides to complex with Ca

1. 1.|The influence of temperature changes on Ca²⁺-binding to brain ganglioside mixtures of different polarity, to single gangliosides (G_M1, G_D1a, G_T1b) and to their deceramide was investigated potentiometrically by means of ion-selective electrodes.

2. 2.|Following cooling (3.5°C/min.) from 37 to 13°C the Ca²⁺-binding to gangliosides, except G_M1, was increased (7–30%).

3. 3.|Subsequent rewarming from 13 to 37°C resulted in up to 100% release of previously-bound Ca²⁺.

4. 4.|When comparing the maximal absolute binding difference of Ca²⁺ to gangliosides during temperature changes a decrease of these differences could be stated which corresponds to an increase in the polarity of the gangliosides.

5. 5.|From these experiments it is concluded that a higher polarity of neuronal gangliosides is responsible for a lower thermal sensitivity of Ca²⁺-binding to these compounds. This may be involved in the process of thermal adaptation of ectothermic vertebrates.

Profiles of steroids during third trimester of pregnancy in women: Effect of hot, dry desert environment

1. 1.|Profile of steroids were determined in either serum or urine of nonparous desert dwelling women, in summer (June–September) and in winter (December–February), both during the latter part of pregnancy or the luteal phase of the menstrual cycle.

2. 2.|Progesterone concentration was consistently lower in serum of women having their third trimester of pregnancy during summer. No such changes were noted in the concentration of unconjugated oestriol.

3. 3.|The urinary excretion rate of glucocorticoids, mineralocorticoids and their metabolites by women in week 35 of pregnancy during summer differed from that excreted by women during winter.

4. 4.|None of the changes in the profiles of steroids attributable to the natural thermal load of the summer season was associated with abrogation of pregnancy and foetal distress.

Thermosensitivity during embryonic development of Lymnaea stagnalis (Mollusca)

1. 1.|The percentage of survival after 1 hr at 40.0°C is lowest at the larval trochophore stage and at hatching of the young snail.

2. 2.|Heat resistance depends on the stage of development.

3. 3.|From the early cleavage stage onwards a higher percentage of embryos can withstand high temperature after a previous heat treatment than without it.

4. 4.|The pattern of thermosensitivity is discussed in relation to the organizational level of the stage of development.

5. 5.|It is concluded that the developing Lymnaea is a suitable system to study heat resistance and thermotolerance at the level of cells, organs and organism.

Heat-transfer relations of avian nestlings

1. 1.|To determine the thermoregulatory prowess of altricial nestlings, we conducted both equilibrium and transient analyses of white-crowned sparrow nestings, a representative fringillid.

2. 2.|For an individual nestling at thermal equilibrium, feather development is the major factor reducing heat loss after 2 days of age; tissue- and boundary-layer resistances are of minor importance.

3. 3.|The nest substantially reduces wind speeds near the nestlings. Heat transfer through the nest material is of only moderate importance. Evaporation also appears to be a small proportion of total heat loss during hypothermia in natural environments.

4. 4.|Net long-wave radiant exchange is also minor, but short-wave radiation is potentially a major component of the nestling's energy budget, approaching the magnitude of maximal metabolic heat production.

5. 5.|When nestlings cool, their body mass and metabolic rate are also major importance in determining the rate of cooling, and (for metabolism) the equilibrium temperature as well.

6. 6.|The huddling together of nestlings is perhaps the single most important factor affecting heat transfer.

7. 7.|An older brood actually has more insulation than does an adult in the same microclimate.

Employment of a turbidimetric assay system to measure heat-induced protein aggregation

1. 1.|We developed a turbidimetric assay system for quantitation of heat-induced protein aggregation which is presumably caused by protein denaturation.

2. 2.|Rhodanese in 6 M guanidinium chloride was employed in the assay system, because this protein recognizes hydrophobic sites on denatured proteins and aggregates.

3. 3.|Turbidity caused by protein-rhodanase aggregation was recorded at 320 nm by using a u.v./VIS spectrophotometer.

4. 4.|When heated, alcohol dehydrogenase (ADH) aggregates with rhodanese. The increase of ADH-rhodanese aggregation was correlated with the loss of enzymatic activity.

5. 5.|These results indicated that the aggregation was proportional to the extent of ADH denaturation which assumingly caused the loss of ADH activity during heating at 45.5°C.

6. 6.|Similar results were observed when cytosolic proteins from CHO cells were heated at 45.5°C. Heated cytosolic proteins promoted aggregation by complex formation with rhodanese. The aggregation increased with increasing heat dose.

7. 7.|Therefore, the rhodanese assay system can be employed usefully to quantitate the protein aggregation after heat stress.

The effect of temperature changes on the spontaneous activity in the neural ganglia of the cockroach, Periplaneta americana

1. 1.|The effect of localized heating or protocerebrum (bp), the prothoracic (t₁), metathoracic (t₃) and the sixth abdominal (a₆) ganglion on the spontaneous neuronal firing rate in these ganglia was investigated in Periplaneta americana.

2. 2.|In almost every case heating the ganglion increased the firing rate. The most heat sensitive were bp, t₃ and t₁, a₆ was much less so.

3. 3.|In bp and t₁ the firing rate stayed on an increased level even 90 s after the thermal stimulus was over.

4. 4.|The differentiated thermosensitivity of the tested ganglia is discussed in terms of thermoregulatory behaviour of the cockroach.

Electrical skin resistance and rectal temperature changes in resting human subjects exposed to heat

1. 1.|Fourteen male volunteers were examined under passive heating.

2. 2.|Electrical skin resistance (ESR) and rectal temperature (T_re) were measured during the whole period of exposure.

3. 3.|It was found that:

• —|ESR decreases rapidly with increasing air temperature. Assuming an exponential curve yields a mean time constant of 14 min.

• —|There is a correlation between the individual ESR time constants and T_re increases (r = 0.695, P < 0.005).

• —|Additional changes of ESR were noted in 8 subjects at a constant air temperature of 42°C.

4. 4.|It is concluded that ESR may be a useful indicator of the sweating response of the human thermoregulatory system during exogenous heat load.

Acclimatory responses in enzymatic activity and metabolite concentrations in the tropical scorpion Heterometrus fulvipes

1. 1.|Changes in tissue metabolite concentrations and enzyme activities in the pedipalpal (PM) and heart (HM) muscles of the tropical scorpion Heterometrus fulvipes show that the metabolism in PM and HM is fundamentally reorganized following low (18°C) and high (38°C) temperature acclimation.

2. 2.|Changes in metabolite concentrations show that metabolite biosynthesis showed increases after cold acclimation but decreases after warm acclimation.

3. 3.|Similarly, changes in enzyme activities show a preponderance of glycolysis and HMP shunt activity after cold acclimation, while after warm acclimation glycogenolysis, oxidative metabolism and gluconeogenesis predominated.

4. 4.|Higher metabolite concentrations and enzyme activities both before and after thermal acclimation in HM reflect its greater compensatory abilities.