Oxygen limitation of thermal tolerance in cod, Gadus morhua L., studied by magnetic resonance imaging and on-line venous oxygen monitoring

The hypothesis of an oxygen-limited thermal tolerance due to restrictions in cardiovascular performance at extreme temperatures was tested in Atlantic cod, Gadus morhua (North Sea). Heart rate, changes in arterial and venous blood flow, and venous oxygen tensions were determined during an acute temperature change to define pejus ("getting worse") temperatures that border the thermal optimum range. An exponential increase in heart rate occurred between 2 and 16 degrees C (Q(10) = 2.38 +/- 0.35). Thermal sensitivity was reduced beyond 16 degrees C when cardiac arrhythmia became visible. Flow-weighted magnetic resonance imaging (MRI) measurements of temperature-dependent blood flow revealed no exponential but a hyperbolic increase of blood flow with a moderate linear increase at temperatures >4 degrees C. Therefore, temperature-dependent heart rate increments are not mirrored by similar increments in blood flow. Venous Po(2) (Pv(O(2))), which reflects the quality of oxygen supply to the heart of cod (no coronary circulation present), followed an inverse U-shaped curve with highest Pv(O(2)) levels at 5.0 +/- 0.2 degrees C. Thermal limitation of circulatory performance in cod set in below 2 degrees C and beyond 7 degrees C, respectively, characterized by decreased Pv(O(2)). Further warming led to a sharp drop in Pv(O(2)) beyond 16.1 +/- 1.2 degrees C in accordance with the onset of cardiac arrhythmia and, likely, the critical temperature. In conclusion, progressive cooling or warming brings cod from a temperature range of optimum cardiac performance into a pejus range, when aerobic scope falls before critical temperatures are reached. These patterns might cause a shift in the geographical distribution of cod with global warming.     

Effects of seasonal and latitudinal cold on oxidative stress parameters and activation of hypoxia inducible factor (HIF-1) in zoarcid fish

Acute, short term cooling of North Sea eelpout Zoarces viviparus is associated with a reduction of tissue redox state and activation of hypoxia inducible factor (HIF-1) in the liver. The present study explores the response of HIF-1 to seasonal cold in Zoarces viviparus, and to latitudinal cold by comparing the eurythermal North Sea fish to stenothermal Antarctic eelpout (Pachycara brachycephalum). Hypoxic signalling (HIF-1 DNA binding activity) was studied in liver of summer and winter North Sea eelpout as well as of Antarctic eelpout at habitat temperature of 0°C and after long-term warming to 5°C. Biochemical parameters like tissue iron content, glutathione redox ratio, and oxidative stress indicators were analyzed to see whether the cellular redox state or reactive oxygen species formation and HIF activation in the fish correlate. HIF-1 DNA binding activity was significantly higher at cold temperature, both in the interspecific comparison, polar vs. temperate species, and when comparing winter and summer North Sea eelpout. Compared at the low acclimation temperatures (0°C for the polar and 6°C for the temperate eelpout) the polar fish showed lower levels of lipid peroxidation although the liver microsomal fraction turned out to be more susceptible to lipid radical formation. The level of radical scavenger, glutathione, was twofold higher in polar than in North Sea eelpout and also oxidised to over 50%. Under both conditions of cold exposure, latitudinal cold in the Antarctic and seasonal cold in the North Sea eelpout, the glutathione redox ratio was more oxidised when compared to the warmer condition. However, oxidative damage parameters (protein carbonyls and thiobarbituric acid reactive substances (TBARS) were elevated only during seasonal cold exposure in Z. viviparus. Obviously, Antarctic eelpout are keeping oxidative defence mechanisms high enough to avoid accumulation of oxidative damage products at low habitat temperature. The paper discusses how HIF could be instrumental in cold adaptation in fish.     

Effects of acute heat stress and subsequent stress removal on function of hepatic mitochondrial respiration,ROS production and lipid peroxidation in broiler chickens

In order to investigate the effects of acute heat stress and subsequent stress removal on function of hepatic mitochondrial respiration, production of reactive oxygen species (ROS) and lipid peroxidation in broiler chickens, 128 six-week-old broiler chickens were kept in a controlled-environment chamber. The broiler chickens were initially kept at 25 °C (relative humidity, RH, 70 ± 5%) for 6 d and subsequently exposed to 35 °C (RH, 70 ± 5%) for 3 h, then the heat stress was removed and the temperature returned to 25 °C (RH, 70 ± 5%). Blood and liver samples were obtained before heat exposure and at 0 (at the end of the three-hour heating episode, this group is also abbreviated as the HT group), 1, 2, 4, 8, 12 h after the stress was removed. The results showed that acute heat stress induced a significant production of ROS, function of the mitochondrial respiratory chain, antioxidative enzymes [superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px)] activity, and formation of malondialdehybe (MDA). Within the first 12 h after removal of the heat stress, the acute modification of the above parameters induced by heat stress gradually approached to pre-heat levels. The results of the present study suggest that acute exposure to high temperatures may depress the activity of the mitochondrial respiratory chain. This leads to over-production of ROS, which ultimately results in lipid peroxidation and oxidative stress. When the high temperature was removed, the production of ROS, mitochondrial respiratory function and oxidative injury that were induced by acute heat exposure gradually approached the levels observed before heating, in a time-dependent manner.     

Metabolic demand,oxygen supply,and critical temperatures in the Antarctic bivalve Laternula elliptica

Oxygen consumption (Mo(2)), heartbeat rate and form, and circulating hemolymph oxygen content were measured in relation to temperature in the large Antarctic infaunal bivalve Laternula elliptica. After elevations in temperature from 0 degrees to 3 degrees, 6 degrees, and then 9 degrees C, Mo(2) and heartbeat rate rose to new levels, whereas maximum circulating hemolymph oxygen content fell. At 0 degrees C, Mo(2) was 19.6 micromol O(2) h(-1) for a standard animal of 2-g tissue ash-free dry mass, which equates to a 8.95-g tissue dry-mass or 58.4-g tissue wet-mass animal. Elevation of metabolism following temperature change had acute Q(10) values between 4.1 and 5, whereas acclimated figures declined from 3.4 (between 0 degrees and 3 degrees C) to 2.2 (3 degrees -6 degrees C) and 1.9 (6 degrees -9 degrees C). Heartbeat rate showed no acclimation following temperature elevations, with Q(10) values of 3.9, 3.2, and 4.3, respectively. Circulating hemolymph oxygen content declined from 0 degrees to 3 degrees and 6 degrees C but stayed at a constant Po(2) (73-78 mmHg) and constant proportion ( approximately 50%) of the oxygen content of the ambient water. At 9 degrees C, Mo(2) and heartbeat rate both peaked at values 3.3 times those measured at 0 degrees C, which may indicate aerobic scope in this species. After these peaks, both measures declined rapidly over the ensuing 5 d to the lowest measured in the study, and the bivalves began to die. Hemolymph oxygen content fell dramatically at 9 degrees C to values between 2% and 12% of ambient water O(2) content and had a maximum Po(2) of around 20 mmHg. These data indicate an experimental upper lethal temperature of 9 degrees C and a critical temperature, where a long-term switch to anaerobic metabolism probably occurs, of around 6 degrees C for L. elliptica. Concurrent measures of mitochondrial function in the same species had indicated strong thermal sensitivity in proton leakage costs, and our data support the hypothesis that as temperature rises, mitochondrial maintenance costs rapidly outstrip oxygen supply mechanisms in cold stenothermal marine species.     

Cold acclimation allows Drosophila flies to maintain mitochondrial functioning under cold stress

Environmental stress generally disturbs cellular homeostasis. Researchers have hypothesized that chilling injury is linked to a shortage of ATP. However, previous studies conducted on insects exposed to nonfreezing low temperatures presented conflicting results. In this study, we investigated the mitochondrial bioenergetics of Drosophila melanogaster flies exposed to chronic cold stress (4 °C). We assessed mitochondrial oxygen consumption while monitoring the rate of ATP synthesis at various times (0, 1, 2, and 3 days) during prolonged cold stress and at two assay temperatures (25 and 4 °C). We compared organelle responses between cold-susceptible and cold-acclimated phenotypes. Continuous exposure to low temperature provoked temporal declines in the rates of mitochondrial respiration and ATP synthesis. Respiratory control ratios (RCRs) suggested that mitochondria were not critically uncoupled. Nevertheless, after 3 days of continuous cold stress, a sharp decline in the mitochondrial ATP synthesis rate was observed in control flies when they were assayed at low temperature. This change was associated with reduced survival capacity in control flies. In contrast, cold-acclimated flies exhibited high survival and maintained higher rates of mitochondrial ATP synthesis and coupling (i.e., higher RCRs). Adaptive changes due to cold acclimation observed in the whole organism were thus manifested in isolated mitochondria. Our observations suggest that cold tolerance is linked to the ability to maintain bioenergetics capacity under cold stress.     

Effects of temperature on modulation of oxidative stress and antioxidant defenses in testes of tropical tasar silkworm Antheraea mylitta

High temperatures are known to cause physiological stress in organisms. This is often associated with enhanced generation of reactive oxygen species (ROS) leading to oxidative damage. The commercially important tropical tasar silkworm Antheraea mylitta has to endure high summer temperature before egg production on the onset of monsoon. In this study the status of pro-oxidants and antioxidants was studied in the testes of male pupae of tasar silkworm A. mylitta under thermal stress condition. Further, to find out the impact of temperature on physiological activity, oxygen consumption rate was measured. The result indicated higher level of thiobarbituric acid reactive substances (TBARS, as an index of lipid peroxidation) and total hydroperoxides in the male pupae exposed to high temperature (40±1 °C). Similarly, it was found that increased levels of antioxidants such as superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), ascorbic acid (ASA) and low molecular thiols (L-SH) in testes are more prominent in high temperature rather than in moderate temperature (35±1 ^oC) suggesting the activation of physiological mechanism to scavenge the ROS produced during stress. Further more, the order of higher level of oxygen consumption rate was observed as high temperature (40±1°C) > moderate temperature (35±1°C) > control groups (28±1°C). Oxygen consumption rate was positively correlated with oxidative stress and antioxidant defence indices. We infer from these findings that the testes of A. mylitta pupae modulate testicular antioxidant responses to thermal stress.     

Oxygen-limited thermal tolerance in Antarctic fish investigated by MRI and (31)P-MRS

The hypothesis of an oxygen-limited thermal tolerance was tested in the Antarctic teleost Pachycara brachycephalum. With the use of flow-through respirometry, in vivo (31)P-NMR spectroscopy, and MRI, we studied energy metabolism, intracellular pH (pH(i)), blood flow, and oxygenation between 0 and 13 degrees C under normoxia (PO(2): 20.3 to 21.3 kPa) and hyperoxia (PO(2): 45 kPa). Hyperoxia reduced the metabolic increment and the rise in arterial blood flow observed under normoxia. The normoxic increase of blood flow leveled off beyond 7 degrees C, indicating a cardiovascular capacity limitation. Ventilatory effort displayed an exponential rise in both groups. In the liver, blood oxygenation increased, whereas in white muscle it remained unaltered (normoxia) or declined (hyperoxia). In both groups, the slope of pH(i) changes followed the alpha-stat pattern below 6 degrees C, whereas it decreased above. In conclusion, aerobic scope declines around 6 degrees C under normoxia, marking the pejus temperature. By reducing circulatory costs, hyperoxia improves aerobic scope but is unable to shift the breakpoint in pH regulation or lethal limits. Hyperoxia appears beneficial at sublethal temperatures, but no longer beyond when cellular or molecular functions become disturbed.     

Effects of salinity and cyclic temperature on survival of two sympatric species of grass shrimp (Palaemonetes), and their relationship to natural distributions

Survival and respiration of the grass shrimp Palaemonetes pugio Holthuis and P. vulgaris (Say) from the Newport River estuary were measured after exposure to cyclic and constant winter temperatures, to rapid decreases in temperature, and to various temperature-salinity combinations. Both species were subjected to nine temperature-salinity combinations including temperature regimes of cyclic 7–13°C, constant 7° and 10°C, and salinities of 5, 20, and 35%.. Based on the laboratory and field results, the differences in physiological tolerance to winter temperatures and salinities were examined in relation to habitat partitioning by these sympatric species.Survival after continuous exposure to cyclic temperature regimes at medium to high salinities was similar to that observed for comparable constant temperatures; however, at low salinities mortality was significantly lower under the cyclic regime than under either constant regime. This suggests that cyclic temperatures may be detrimental in combination with some other stress. A rapid, transient decrease in temperature from either 7° or 10°C to 2°C had no measurable effect on survival or rate of oxygen consumption at any temperature-salinity acclimation. Neither salinity (except in areas intermittently subjected to salinities below 3 %.) nor winter temperatures appear to affect habitat partitioning in grass shrimp.     

Oxygen limitation of thermal tolerance defined by cardiac and ventilatory performance in spider crab, Maja squinado

Geographic distribution limits of ectothermal animals appear to be correlated with thermal tolerance thresholds previously identified from the onset of anaerobic metabolism. Transition to these critical temperatures was investigated in the spider crab (Maja squinado) with the goal of identifying the physiological processes limiting thermal tolerance. Heart and ventilation rates as well as PO(2) in the hemolymph were recorded on-line during progressive temperature change between 12 and 0 degrees C (1 degrees C/h) and between 12 and 40 degrees C (2 degrees C/h). Lactate and succinate were measured in tissues and hemolymph after intermediate or final temperatures were reached. High levels of hemolymph oxygenation suggest that an optimum range of aerobic performance exists between 8 and 17 degrees C. Thermal limitation may already set in at the transition from optimum to pejus (pejus = turning worse, progressively deleterious) range, characterized by the onset of a decrease in arterial PO(2) due to reduced ventilatory and cardiac performance. Hemolymph PO(2) values fell progressively toward both low and high temperature extremes until critical temperatures were reached at approximately 1 and 30 degrees C, as indicated by low PO(2) and the onset of anaerobic energy production by mitochondria. In conclusion, the limited capacity of ventilation and circulation at extreme temperatures causes insufficient O(2) supply, thereby limiting aerobic scope and, finally, thermal tolerance.     

Effect of thermoperiod on diapause intensity in Pyrrhocoris apterus (Heteroptera Pyrrhocoridae)

The intensity of adult diapause in Pyrrhocoris apterus was measured in two series of experiments as the duration of pre-oviposition period at a constant temperature of 25 degrees C after transfer from short (12L:12D) to long day conditions (18L:6D). Higher diapause intensity was induced with a thermoperiod than at constant temperatures. After the induction throughout larval instars 3-5 and during 4 weeks of adult life at short days and a thermoperiod of 25/15 degrees C the pre-oviposition period was 30+/-4 and 26+/-3 days. After induction at constant 25 degrees C the pre-oviposition period was 22+/-3 and 23+/-4 days, while after induction at constant 20 degrees C it was 17+/-4 and 19+/-4 days. Induction at a lower constant temperature of 20 degrees C was thus followed by a less intense diapause than the induction at a higher constant temperature of 25 degrees C. These counterintuitive results are discussed. The oxygen consumption rate measured at experimental temperatures prior to transfer from short to long days was higher at thermoperiodic conditions than at constant temperatures and it was similar at constant 20 and 25 degrees C. Thus, the oxygen consumption rate measured prior to the transfer was highest (indication of the least intense diapause) in the insects that showed later, after the transfer to long days, the longest pre-oviposition period (indication of the most intense diapause). Within the first two days after transfer to constant 25 degrees C, oxygen consumption rate measured at 25 degrees C decreased in the thermoperiodic insects, while it transiently increased in insects from constant 20 degrees C. Two days and later after the transfer, oxygen consumption rate was similar in all groups. Cold hardiness was not correlated with diapause intensity. The low lethal temperature in diapausing insects was correlated with the night temperature during diapause induction.     

The impact of the thermal sensitivity of cytochrome c oxidase on the respiration rate of Arctic charr red muscle mitochondria

To assess if cytochrome c oxidase could determine the response of mitochondrial respiration to changes in environmental temperature in ectotherms, we performed KCN titration of the respiration rate and cytochrome c oxidase activity in mitochondria from Arctic charr (Salvelinusfontinalis) muscle at four different temperatures (1 degrees C, 6 degrees C, 12 degrees C, and 18 degrees C). Our data showed an excess of cytochrome c oxidase activity over the mitochondrial state 3 respiration rate. Mitochondrial oxygen consumption rates reached approximately 12% of the cytochrome c oxidase maximal capacity at every temperature. Also, following titration, the mitochondrial respiration rate significantly decreased when KCN reached concentrations that inhibit almost 90% of the cytochrome c oxidase activity. This strongly supports the idea that the thermal sensitivity of the maximal mitochondrial respiration rate cannot be dictated by the effect of temperature on cytochrome c oxidase catalytic capacity. Furthermore, the strong similarity of the Q10s of mitochondrial respiration and cytochrome c oxidase activity suggests a functional or structural link between the two. The functional link could be coevolution of parts of the mitochondrial system to maintain optimal functions in most of the temperature range encountered by organisms.     

Can phenotypic plasticity facilitate the geographic expansion of the tilapia Oreochromis mossambicus?

Climate influences the distribution of organisms because of the thermal sensitivity of biochemical processes. Animals may compensate for the effects of variable temperatures, and plastic responses may facilitate radiation into different climates. The tropical fish Oreochromis mossambicus has radiated into climates that were thought to be thermally unsuitable. Here, we test the hypothesis that thermal acclimation will extend the locomotory and metabolic performance range of O. mossambicus. Juvenile fish were acclimated to 14 degrees, 17 degrees, and 22 degrees C. We measured responses to acclimation at three levels of organization: whole-animal performance (sustained swimming and resting and recovery rates of oxygen consumption), mitochondrial oxygen consumption in caudal muscle, and metabolic enzyme activities in muscle and liver at 12 degrees, 14 degrees, 17 degrees, 22 degrees, and 26 degrees C. Thermal optima of sustained swimming performance (U(crit)) changed significantly with acclimation, but acclimation had no effect on either resting or recovery oxygen consumption. Fish compensated for cold temperatures by upregulating state 3 mitochondrial oxygen consumption and increasing activity of lactate dehydrogenase in the liver. The capacity for phenotypic plasticity in O. mossambicus means that the fish would not be limited by its locomotor performance or metabolic physiology to expand its range into cooler thermal environments from its current distribution.     

Intracellular pH and energy metabolism in the highly stenothermal Antarctic bivalve Limopsis marionensis as a function of ambient temperature

Changes in oxygen consumption, ammonia excretion and in the acid-base and energy status of various tissues were investigated in the cold stenothermal Antarctic bivalve, Limopsis marionensis, and compared to similar data in the limpet, Nacella concinna, for an assessment of thermal sensitivity. Oxygen consumption of L. marionensis varied between −1.5 and 2°C with a Q ₁₀ of 2.2. Ammonia excretion could only be detected in animals exposed to elevated temperature for periods in excess of 45 days and close to death and it is interpreted as the onset of protein and amino acid catabolism with starvation under temperature stress. In L. marionensis any change in temperature as well as starvation stress at constant temperature induced a decrease in phospho-l-arginine and ATP levels. However, only temperature stress resulted in a drop in the Gibb's free energy change of ATP hydrolysis. Intracellular pH rose in all tissues during upward or downward temperature changes of only 1.5 or 2°C for 24 h with a concomitant trend to accumulate succinate and acetate in the tissues. These changes are seen to reflect disturbances of the tissue acid-base and energy status with any under- or overshoot in aerobic metabolic rate during a temperature decrease or increase. Elevated temperature at 2°C during 2 weeks of incubation resulted in continued net ATP depletion, at low levels of ATP free energy. This indicates long-term stress, which was also mirrored in the inability to establish a new steady-state mean rate of oxygen consumption. Incubation at even higher temperatures of 4 and 7°C led to an aggravation of energetic stress and transition to an intracellular acidosis, as well as a fall in oxygen consumption. In N. concinna a drop in energy levels was also visible at 2°C but was compensated for during long-term incubation. In conclusion, L. marionensis will be able to compensate for a temperature change only in a very narrow range whereas the thermal tolerance window is much wider in N. concinna. The inability of the metabolic rate to rise continually and the concomitant transition to anaerobic metabolism and long-term energetic stress characterize the upper critical temperature. Stenothermality is discussed, not only as reflecting the permanent and very stable low temperature in the natural environment, but also regarding dif- ferences in the level of activity and aerobic scope. Accepted: 21 December 1998     

Longevity of animals under reactive oxygen species stress and disease susceptibility due to global warming

The world is projected to experience an approximate doubling of atmospheric CO_2 concentration in the next decades. Rise in atmospheric CO_2 level as one of the most important reasons is expected to contribute to raise the mean global temperature 1.4 ℃-5.8 ℃ by that time. A survey from 128 countries speculates that global warming is primarily due to increase in atmospheric CO_2 level that is produced mainly by anthropogenic activities. Exposure of animals to high environmental temperatures is mostly accompanied by unwanted acceleration of certain biochemical pathways in their cells. One of such examples is augmentation in generation of reactive oxygen species(ROS) and subsequent increase in oxidation of lipids, proteins and nucleic acids by ROS. Increase in oxidation of biomolecules leads to a state called as oxidative stress(OS). Finally, the increase in OS condition induces abnormality in physiology of animals under elevated temperature. Exposure of animals to rise in habitat temperature is found to boost the metabolism of animals and a very strong and positive correlation exists between metabolism and levels of ROS and OS. Continuous induction of OS is negatively correlated with survivability and longevity and positively correlated with ageing in animals. Thus, it can be predicted that continuous exposure of animals to acute or gradual rise in habitat temperature due to global warming may induce OS, reduced survivability and longevity in animals in general and poikilotherms in particular. A positive correlation between metabolism and temperature in general and altered O_2 consumption at elevated temperature in particular could also increase the risk of experiencing OS in homeotherms. Effects of global warming on longevity of animals through increased risk of protein misfolding and disease susceptibility due to OS as the cause or effects or both also cannot be ignored. Therefore, understanding the physiological impacts of global warming in relation to longevity of animals will become very crucial challenge to biologists of the present millennium.     

Silver Nanoparticle Exposure Induced Mitochondrial Stress,Caspase-3 Activation and Cell Death: Amelioration by Sodium Selenite

Silver nanoparticles (AgNP), one of the most commonly used engineered nanomaterial for biomedical and industrial applications, has shown a toxic potential to our ecosystems and humans. In this study, murine hippocampal neuronal HT22 cells were used to delineate subcellular responses and mechanisms to AgNP by assessing the response levels of caspase-3, mitochondrial oxygen consumption, reactive oxygen species (ROS), and mitochondrial membrane potential in addition to cell viability testing. Selenium, an essential trace element that has been known to carry protecting property from heavy metals, was tested for its ameliorating potential in the cells exposed to AgNP. Results showed that AgNP reduced cell viability. The toxicity was associated with mitochondrial membrane depolarization, increased accumulation of ROS, elevated mitochondrial oxygen consumption, and caspase-3 activation. Treatment with sodium selenite reduced cell death, stabilized mitochondrial membrane potential and oxygen consumption rate, and prevented accumulation of ROS and activation of caspase-3. It is concluded that AgNP induces mitochondrial stress and treatment with selenite is capable of preventing the adverse effects of AgNP on the mitochondria.     

Temperature-dependent energy allocation to growth in Antarctic and boreal eelpout (Zoarcidae)

Antarctic fishes display slower annual growth rates than congeneric species from temperate zones. For an analysis of growth in relation to energy turnover, body composition was analysed in two benthic fish species to establish a whole animal energy budget. The Antarctic eelpout, Pachycara brachycephalum, was maintained at 0, 2, 4 and 6°C and the boreal eelpout, Zoarces viviparus at 4, 6, 12 and 18°C. At maximum food supply the weight gain was highest for P. brachycephalum at 4°C. Routine metabolic rate in acclimated Antarctic eelpouts did not differ between temperatures, whereas in Z. viviparus maximized growth benefited from a reduction of metabolic energy demands at 12°C. The lipid content of liver declined with increasing temperature in both species. The thermal window for growth is based on food conversion efficiency and the level of metabolic energy demand and is limited according to the level of aerobic scope available between pejus temperatures.     

Mitochondrial metabolic suppression in fasting and daily torpor: consequences for reactive oxygen species production

Abstract Daily torpor results in an ～70% decrease in metabolic rate (MR) and a 20%-70% decrease in state 3 (phosphorylating) respiration rate of isolated liver mitochondria in both dwarf Siberian hamsters and mice even when measured at 37°C. This study investigated whether mitochondrial metabolic suppression also occurs in these species during euthermic fasting, when MR decreases significantly but torpor is not observed. State 3 respiration rate measured at 37°C was 20%-30% lower in euthermic fasted animals when glutamate but not succinate was used as a substrate. This suggests that electron transport chain complex I is inhibited during fasting. We also investigated whether mitochondrial metabolic suppression alters mitochondrial reactive oxygen species (ROS) production. In both torpor and euthermic fasting, ROS production (measured as H(2)O(2) release rate) was lower with glutamate in the presence (but not absence) of rotenone when measured at 37°C, likely reflecting inhibition at or upstream of the complex I ROS-producing site. ROS production with succinate (with rotenone) increased in torpor but not euthermic fasting, reflecting complex II inhibition during torpor only. Finally, mitochondrial ROS production was twofold more temperature sensitive than mitochondrial respiration (as reflected by Q(10) values). These data suggest that electron leak from the mitochondrial electron transport chain, which leads to ROS production, is avoided more efficiently at the lower body temperatures experienced during torpor.     

Impacts of fluctuating temperature on the development and infectivity of entomopathogenic nematode Steinernema carpocapsae A10

Three different laboratory conditions were used to examine the impacts of fluctuating temperature on the development and infectivity of entomopathogenic nematode (EPN) Steinernema carpocaposae A10. Set I experiments focused on the impact of cold stress early in the development cycle. In these studies Galleria mellonella hosts were infected and incubated for 2 days at the control temperature of 23 degrees C and then subjected to lower temperatures of -10, 4, 10 or 14 degrees C, respectively, from days 3 to 36 post-infection (PI). Dissections of infected cadavers indicated arrested development at the adult stage at all lower temperatures tested. Set II experiments examined the impacts of cold stress early in the development followed by a return to 23 degrees C. Hosts were infected and incubated as in Set I and subjected to the same temperatures as above for 7 days, followed by incubation at 23 degrees C until 23 days PI. A limited number of EPN populations were able to complete development at 10 and 14 degrees C though emergent population numbers were significantly lower than those of control infections incubated continuously at 23 degrees C. In Set III experiments, infected hosts were subjected to cold stress later during development starting at day 4 post-infection followed by incubation at the control temperature. Population survival past first and second stage juveniles was reduced by at least 95% or more at the lower temperatures compared with controls. Emergent populations from the Set III cold-stressed hosts were not infectious. These studies may provide insights as to how EPN survive seasonal temperature fluctuations under natural environmental conditions.     

Synaptosomal response to oxidative stress: Effect of vinpocetine

It has been suggested that reactive oxygen species (ROS) play a role in the neuronal damage occurring in ischemic injury and neurodegenerative disorders and that their neutralization by antioxidant drugs may delay or minimize neurodegeneration. In the present study we examine whether vinpocetine can act as an antioxidant and prevent the formation of ROS and lipid peroxidation in rat brain synaptosomes. After ascorbate/Fe²⁺ treatment a significant increase in oxygen consumption (about 5-fold) and thiobarbituric acid reactive substances (TBARS) formation (about 7-fold) occurred as compared to control conditions. Vinpocetine inhibited the ascorbate/Fe²⁺ stimulated consumption of oxygen and TBARS accumulation, an indicator of lipid peroxidation, in a concentration-dependent manner. The ROS formation was also prevented by vinpocetine. Oxidative stress increased significantly the fluorescence of the probes 2′,7′-dichlorodihydrofluorescein (DCFH₂-DA) (about 6-fold) and dihydrorhodamine (DHR) 123 (about 10-fold), which is indicative of intrasynaptosomal ROS generation. Vinpocetine at 100 μM concentration decreased the fluorescence of DCFH₂-DA and DHR 123 by about 50% and 83%, respectively. We conclude that the antioxidant effect of vinpocetine might contribute to the protective role exerted by the drug in reducing neuronal damage in pathological situations.