The role of ethylene in the prevention of chilling injury in nectarines

Woolliness is a chilling injury phenomenon occurring in nectarines held at low temperatures for extended periods. It is a disorder marked by altered cell wall metabolism during ripening leading to a dry, woolly texture in the fruit. Two treatments were found to alleviate this disorder. One was holding the fruits for 2 days at 20 °C before 0 °C storage (delayed storage) and the second was having ethylene present during cold storage (ethylene). Immediately stored fruit (control) had 88 percnt; woolliness while 7 percnt; of delayed storage and 15 percnt; of ethylene fruit showed woolliness. The severity of the injury in individual fruits was closely related to inhibition of ethylene evolution. Woolly fruit had higher levels of 1-aminocyclopropane-1-carboxylic acid (ACC) and less 1-aminocyclopropane-1-carboxylic acid oxidase (ACO, EC 1.4.3) activity than healthy fruit. It is suggested that ethylene is essential for promoting the proper sequence of cell wall hydrolysis necessary for normal fruit softening. This is in contrast to chilling injury in other fruits, whereby ethylene is often a sign of incipient damage. Respiration was also found to be associated with chilling injury, in that fruit with woolliness had a depressed respiration.     

Chilling Injury,Physicochemical Properties,and Antioxidant Enzyme Activities of Red Pitahaya (<i>Hylocereus polyrhizus</i>) Fruits under Cold Storage Stress

Low-temperature storage is extensively used to optimize the postharvest life of various fresh fruits. However, red pitahaya (Hylocereus polyrhizus) fruits are sensitive to chilling injury (CI), which leads to the limitation of low-temperature storage. In this study, red pitahaya fruits were stored at 2, 4, 6, 8, and 10°C, respectively, for 27 days to determine the appropriate storage temperature. During the storage of red pitahaya fruits, storage at 8°C was more effective in suppressing decay and maintaining quality than other low temperatures. Low-temperature (2, 4, and 6°C) storage decreased weight loss (WL) and maintained higher content of titratable acidity (TA), soluble sugars (SS), and total phenolics (TP) but different degrees of CI were detected. No CI was observed at 8°C and 10°C. Red pitahay as stored at 8 and 10°C were associated with better color evaluation, lower electrolyte leakage (EL), respiration rate, and lipoxygenase (LOX) activity, and higher fruit firmness, superoxide dismutase (SOD) activity, and catalase (CAT) activity. However, higher storage temperature (10°C) resulted in higher metabolic activity leading to lower quality and antioxidant capacities compared with 8°C. Therefore, our results demonstrated that red pitahaya stored at 8°C exhibited a protective effect on fruit quality and resisted CI development during storage.     

Superoxide production by mitochondria isolated from green bell pepper fruit

Evidence is increasing to suggest that a wide range of environmentally induced plant disorders, including chilling injury, is mediated by reactive oxygen species produced during stress or upon relief from stress. Mitochondria were isolated from pericarp tissue of chilling-sensitive bell pepper fruit and their respiratory activity and ability to produce superoxide when supplied with NADH, succinate or malate-pyruvate were determined. Oxygen uptake rates were greater and less sensitive to cyanide with succinate than with NADH; rates increased and sensitivity to cyanide and respiratory control ratios (RCRs) decreased in fruit stored at 2°C. Disrupting mitochondrial membranes led to increased oxygen consumption with NADH and decreased consumption with succinate. resulting in RCRs of approximately 1 with both substrates. Superoxide production was greater with NADH than with either succinate or malatepyruvate. Superoxide dismutase and cyanide inhibited superoxide production almost completely. Antimycin A did not inhibit superoxide production with NADH, but did partially with succinate, especially in mitochondria sensitive to cyanide. Disrupting mitochondrial membranes enhanced superoxide production with NADH. Superoxide production by mitochondria isolated from fruit stored at 2°C increased with NADH and decreased with succinate. Results provide evidence that mitochondria may be a major source of superoxide in chilling-sensitive plant tissues exposed to low temperatures.     

Effects of Chilling Temperatures on Ethylene Binding by Banana Fruit

Banana fruit are highly susceptible to chilling injury during low temperature storage. Experiments were conducted to compare ethylene binding during storage at chilling (3 and 8 °C) versus optimum (13 °C) temperatures. The skins of fruit stored at 3 and 8 °C gradually darkened as storage duration increased. This chilling effect was reflected in increasing membrane permeability as shown by increased relative electrolyte leakage from skin tissue. In contrast, banana fruit stored for 8 days at 13 °C showed no chilling injury symptoms. Exposure of banana fruit to the ethylene binding inhibitor 1-methylcyclopropene (1 l l^-1 1-MCP) prevented ripening. However, this treatment also enhanced the chilling injury accelerated the occurrence of chilling injury-associated increased membrane permeability. ¹⁴C-ethylene release assay showed that ethylene binding by banana fruit stored at low temperature decreased with reduced storage temperature and/or prolonged storage time. Fruit exposed to 1-MCP for 12 h and then stored at 3 or 8 °C exhibited lower ethylene binding than those stored at 13 °C. Thus, chilling injury of banana fruit stored at low temperature is associated with a decrease in ethylene binding. The ability of tissue to respond to ethylene is evidently reduced, thereby resulting in failure to ripen.     

Polyamine content and chilling susceptibility are affected by seasonal changes in temperature and by conditioning temperature in cold-stored 'Fortune' mandarin fruit

‘Fortune’ mandarins are prone to develop pitting and necrosis upon exposure to low temperatures. We have examined the effect of field temperature during fruit maturation and the effect of conditioning temperatures (from 2 to 37°C) prior to cold storage on the content of polyamines (PAs) and on chilling susceptibility in order to understand the role of PAs in maturation and chilling tolerance of this citrus cultivar. Chilling susceptibility and the content of PAs were more affected by seasonal changes in field temperature than by the stage of fruit maturity. The highest putrescine (Put) and spermidine (Spd) content was found in fruits exposed to the lowest field temperatures. These fruits were in turn more susceptible to develop chilling injury (CI) after storage at 2°C. Spermine (Spm), however, decreased in attached fruit with time of exposure to temperatures below 12°C. Temperature pretreatments for 3 days above 20°C of fruits detached from the tree reduced CI, the more so the higher the conditioning temperature. Put and Spd increased with temperature conditioning in detached fruits, differing from the response of fruits attached to the tree. No direct relationship between induced levels of these PAs and the tolerance to CI was found. Levels of Put and Spd increased at temperatures (22, 30 and 37°C) which increased the tolerance and also at temperatures (6 and 12°C) which accelerated the appearance of chilling symptoms. In contrast, a significant increase in Spm levels was only found after conditioning at 30 or 37°C. After cold storage a general decline in PA levels occurred in all temperature‐conditioned mandarins. In most cases no significant differences among fruit exposed to effective and non‐effective pretreatments were observed. PA content increased again after transferring cold‐stored fruits to 20°C, whereas the CI index was barely affected. In conclusion, PA changes in the flavedo of ‘Fortune’ mandarins appear to be related to variations in temperature rather than to stage of maturity or tolerance to chilling.     

Polyamine Changes and Chilling Injury in Cold-stored Loquat Fruits

Free polyamine levels (spermine (SPM), spermidine (SPD), and putrescine (PUT)) were determined using thin-layer chromatography and fluorometric method in loquat (Eriobotrya japonica Lindl. cv. Dahongpao) fruits stored at 1℃ and 12℃ and in postharvest SPM treated fruits stored at 1℃ respectively to investigate the relationship between changes in polyamines and chilling injury. In the loquat fruits stored at 1℃, SPM level decreased gradually in the first two weeks, then increased sharply and reached a peak value after three weeks, thereafter it decreased rapidly. SPD level decreased steadily during the first three weeks and increased significantly afterwards. PUT level evolved in a similar way as the SPM level did except that it increased slowly in the first two weeks. The fruit showed symptom of chilling injury manifested as flesh leatheriness after three weeks. However, no significant increase and decrease of these three polyamines was detected during storage at the nonchilling temperature (12℃). The SPM-treated fruits maintained high levels of SPM and SPD and remained low level of PUT during storage at 1℃, and no symptom of chilling injury was observed. These results suggested that the increase in SPM level in response to chilling exposure might serve as a defense mechanism against chilling injury while the accumulation of PUT could be a cause of the stress-induced injury and the increase in SPD level could be a consequence of this kind of stress.     

Effect of chilling temperatures on the activities of glyoxysomal and mitochondrial enzymes from castor bean seedlings

Castor bean (Ricinus communis L.) is included among the group of plants sensitive to chilling temperatures. Seedlings of this species were shown to exhibit visible symptoms of this injury, as well as impaired radicle growth and storage product mobilization. Respiration of intact seedlings and oxidation of succinate by isolated mitochondria displayed discontinuities in Arrhenius plots of their reaction velocities, characteristic of chilling species. However, gluconeogenic glyoxysomal enzymes do not display such discontinuities, indicating that there is probably no functional relationship between these enzymes and the glyoxysomal membrane.     

Effect of chilling and subsequent storage at 20°C on electrolyte leakage and phospholipid fatty acid composition of tomato pericarp

Mature green tomato fruit ( Lycopersicon esculentum cv. Caruso) were stored at 1°C or 20°C and analyzed on day 0, 18 and 22 for electrolyte leakage, ripening-associated changes in pigmentation and phospholipid fatty acid composition. Chilled fruit were also analyzed 4 days after they were returned to 20°C. Fruit did not ripen significantly during chilling and subsequent storage at 20°C, and showed visible chilling injury symptoms only at 20°C. Electrolyte leakage increased in control and chilled fruit, indicating enhanced membrane permeability during both ripening and chilling. Returning the fruit to ambient temperature gave an apparent decrease in electrolyte leakage. Phospholipid and linolenic acid content and double bond index decreased during ripening at 20°C. The small changes in phospholipid fatty acid composition during chilling cannot account for the enhanced membrane permeability. The significant decrease in percentage of linolenic acid and in double bond index in the total lipids, but not in the phospholipids, upon returning the fruit to 20°C suggests loss of galactolipid polyunsaturated fatty acids     

Effect of fruit type and storage treatments on the biodeterioration of African pear (Dacryodes edulis (G. Don.) H. J. Lam.)

The storage potential of the African pear (Dacryodes edulis (G. Don.) H. J. Lam.) (Burseraceae) was investigated using four fruit types. At ambient temperatures (28·5–30°C), those enclosed in either paper or polythene bags could be stored satisfactorily for 3–8 days, after which they deteriorated rapidly. Storage life was increased at lower temperatures, but injury due to chilling was observed at −5°C. At 15°C, fruits dipped in palm oil before being packaged were of better quality and retained their firmness longer than fruits dipped in a 500 ppm benlate solution. Storage in moist sawdust, wood-shavings or water were the least effective in extending shelf-life. Variation among fruit types in their response to storage treatment was observed.     

Methyl jasmonate treatments reduce chilling injury and activate the defense response of guava fruits

Tropical fruits cannot be stored at low temperatures due to the chilling injury phenomena. With the goal of reducing the chilling injury, we tested 10(-4) and 10(-5) M of methyl jasmonate (MJ) treatment before the storage of red and white cultivars of guava fruits at 5 degrees C for up to 15 days plus two days at 20 degrees C. Every five days, we evaluated chilling injury index, ion leakage percentage, vitamin C, sugars, total phenols, and the activity of the enzymes lipoxygenase (LOX) and phenylalanine-ammonia lyase (PAL). We found that methyl jasmonate treatments reduce the chilling injury index and the ion leakage percentage. Furthermore, MJ did not affect vitamin C, chlorophyll, and total phenols. MJ increased sugar content, PAL, and LOX activities. We concluded that MJ reduces chilling injury and activates the fruit defense response as indicated by the behavior of total phenols and the increase in sugar content, PAL, and LOX activities.     

Photosystem II quantum yield as a measure of radical scavengers in chilling injury in cucumber fruits and bell peppers

Cucumber fruits (Cucumis sativus L. cv. Jessica) and green bell peppers (Capsicum annuum L. cvs. Lokas and Medeo) were stored at different temperatures ranging from 2 to 12°C. After three different storage periods, fruits from each temperature were transferred to 20°C for 7 d to allow for the development of visual symptoms of chilling injury (CI). During storage, the photochemical quantum yield of photosystem II (PSII) in peel tissue adapted to darkness, was calculated from measurements of pulse-amplitude-modulated chlorophyll fluorescence. The decrease in PSII quantum yield during storage at low temperatures in darkness can be described as a temperature-dependent inhibition of an enzyme according to Arrhenius, assuming a negative activation energy. By comparison with the radical-scavenger measurements of Hariyadi and Parkin (1991, Postharvest Biol. Techn.1, 33–45) it is postulated that the time and temperature dependence of the quantum yield parallels the diminution of radical-scavenging activity at lower temperatures in cucumber and capsicum fruits. This is combined with an equation for the process of radical scavenging itself and an equation for the auto-catalytic radical-producing lipid peroxidation reaction. These three basic processes lead to both a static and a dynamic model for the occurrence of chilling injury in low-temperature-sensitive plant tissue. A statistical fit of the measured data using the static model leads to the estimates of the different activation energies and reaction rates with a high degree of accuracy. The estimated values are in accord with what one would expect on the basis of knowledge of the processes leading to chilling injury, and directly point to meaningful physico-chemical parameters.Abbreviations C Symmetry point of logistic curve - CI intensity of chilling injury - e base of the natural logarithm system - E activation energy - Eff efficiency - F_v/F_m quantum yield of PSII - k reaction rate constant - R amount of free radicals - S amount of substrate for chilling injury (double bonds in fatty-acid chains) - t time - T temperature - Z amount of radical-scavenging enzyme Indices - CI chilling injury - d denaturation - i any index - max maximum value - min minimum value - r radical scavenging - ref reference (temperature) - s substrate - 0 initial amount This study was conducted in the framework of a research program on fruits and vegetables, partly financed by the Dutch Commodity Board for Vegetables and Fruits.     

Differential effects of freezing on total hepatic peroxisomal fatty acid oxidation and on the rate-limiting enzyme, acyl-CoA oxidase

Fatty acid oxidation defects can be acutely fatal, leading to the collection of tissues which are frozen for future analysis. Since peroxisomes can also oxidize long-chain fatty acids, differentiation of the contributions from the peroxisome as opposed to the mitochondria is important. We studied the effects of freezing and storage of rat livers on peroxisomal and mitochondrial beta-oxidation as measured by cyanide sensitivity of the oxidation of [1-14C]oleoyl-CoA to 14CO2 and acid-soluble labeled products. In addition, we examined the effects of freezing and storage on the rate-limiting enzyme for peroxisomal beta-oxidation, acyl-CoA oxidase, by the H2O2 generation method. Marked reduction in the oxidation of [1-14C]oleoyl-CoA was found for both peroxisomal and mitochondrial systems upon freezing at -18 or -70 degrees C for 2 days which declined further on storage at these temperatures for 12 weeks. Loss of activity after freezing was greater for the mitochondrial than the peroxisomal beta-oxidation system. By contrast, acyl-CoA oxidase activity was resistant to these changes, maintaining prefrozen activities despite storage for 12 weeks. The contribution of the peroxisomal system to beta-oxidation was 32% of the total rate of oxidation of [1-14C]oleoyl-CoA in the rat liver. These findings indicate that the contributions of the peroxisomal system to total fatty acid oxidation may be considerable, that freezing of the liver results in drastic reduction in enzyme activities of both peroxisomal as well as mitochondrial beta-oxidation, but that the rate-limiting enzyme of the peroxisomal system, acyl-CoA oxidase, retains full activity despite freezing and storage.     

The effect of acclimation temperature on the activation energies of state III respiration and on the unsaturation of membrane lipids of goldfish mitochondria

The influence of the acclimation temperature on the thermotropic behaviour of mitochondrial respiration and on the degree of unsaturation of mitochondrial membrane lipids has been studied. The mitochondria were isolated from red muscle, white muscle and liver of goldfish acclimated to 5, 20 and 30°C. ADP-activated succinate oxidation was measured at different temperatures and resulted in non-linear Arrhenius-plots with breaks between 10 and 23°C. As for the break-temperatures, there was found a shift downwards in preparations of decreased acclimation temperatures. This could be caused by a changed composition of membrane lipids and a simultaneous shift of the membrane phase transition temperature. Therefore, the fatty acid composition of all membrane preparations was analyzed. However, no consistent change of the degree of unsaturation due to a changed acclimation temperature could be found.     

Sorting of an exocrine secretory protein to the regulated secretory pathway in endocrine cells

The effect of exogenous polyamines on electrolyte leakage, chilling index, polygalacturonase activity (PG), ethylene production, and firmness in zucchini squash fruits stored for 12 days at 2 degrees C or 10 degrees C, 85-90% RH was evaluated. Fruits were infiltrated with putrescine (PUT) spermidine (SPD) and spermine (SPM) at 0.1, 0.25, 0.5, 2.0, and 4.0 mM. All polyamines exerted a protective effect on cell and organelle membranes. The most effective was SPD, which reduced electrolyte leakage between 62% and 82%, compared to control fruits stored at 2 degrees C. At 10 degrees C they did not exhibit chilling injury (CI) symptoms, while at 2 degrees C SPM (0.5 mM) and SPD (0.5 mM) diminished them 92% and 100%, respectively; which extended storage life for 8-10 days at 2 degrees C. High concentrations of polyamines (>2.0 mM) caused the appearance of CI symptoms. PG activity diminished proportionally to the concentration of polyamine except for the concentration at 4.0 mM. No significant changes were observed in ethylene production.     

<Emphasis Type="Italic">PpVIN2</Emphasis>, an acid invertase gene family member,is sensitive to chilling temperature and affects sucrose metabolism in postharvest peach fruit

We previously reported that expression and activity of acid invertases (AI) are increased in peach fruit under chilling stress. In order to determine which AI genes respond to chilling stress, seven AI genes, two vacuolar invertases (VINs) and five cell wall-bound invertases (CWINs), were identified and cloned. The predicted amino acid sequences of the genes contain conserved sites characteristic of plant AIs such as NDPNG/A, the sucrose-binding site, and MWECV/P, a cysteine catalytic motif. Using gene-specific primers, the expression of each gene was measured in ‘Baifeng’ and ‘Yulu’ peach fruits stored at 0, 5, 10 and 20 °C. Of the seven genes, expression of PpVIN2 was the most affected by chilling stress; the largest increases were in fruit stored at 5 °C, up to 17-fold in ‘Baifeng’ fruit, and up to 280-fold in ‘Yulu’ fruit. Overall, VIN activity was much higher than CWIN activity in stored peach fruit. In both cultivars reducing sugar content increased significantly and sucrose content decreased gradually during storage at 5 °C relative to other temperatures, and was accompanied by severe chilling injury symptoms. Thus, PpVIN2 appears to be induced by chilling and may play an important role in sucrose metabolism in peach fruit subjected to cold storage.     

Fatty acid oxidation in isolated rat liver mitochondria. Developmental changes and their relation to hepatic levels of carnitine and glycogen and to carnitine acyltransferase activity

Prompted by an apparent relationship between ketosis and fatty acid utilization, we studied the capacities for fatty acid oxidation through β-oxidation and Krebs cycle in liver mitochondria isolated from fetal and suckling rats. Rates of state 3 oxidation, as measured by oxygen consumption, were low for both palmitylcarnitine and palmityl CoA plus carnitine at 2 days before term and at birth, but increased at least ninefold during the first 8 days of life and at least sixfold during the remaining suckling period. Despite these sharp increases, oxygen consumption in suckling rats did not exceed the value for fed adult rats. Also, the rates of state 3 oxidation of succinate were low in suckling rats. Respiratory control indices, determined with each of the three substrates, were lower in suckling rats than fed adults. By contrast, ratios of fatty acyl ester to succinate oxidation, a relative measure of the oxidation of palmitylcarnitine and palmityl CoA, were 21–66% and 27–77% higher in suckling than in fed adult rats. The increased ratios indicate that the capacity for fatty acid oxidation is higher during postnatal development than in the fetal stage or adulthood. The oxidation capacity was inversely related to glycogen content in the liver. Although hepatic carnitine concentration and carnitine palmityltransferase activity increased during suckling period, they are not rate limiting for fatty acid oxidation. Studies of the partitioning of fatty acids showed that about two-thirds of the fatty acid oxidized through β-oxidation did not enter Krebs cycle for further oxidation. These results support our working hypothesis that ketosis of suckling rats stems from rapid oxidation of fatty acids and increased partitioning of acetyl CoA into ketogenesis.     

Effects of paclobutrazol and chilling temperatures on lipids, antioxidants and ATPase activity of plasma membrane isolated from green bell pepper fruits

The effects of paclobutrazol treatment on plasma membrane lipid composition and ATPase activity of bell pepper fruit ( Capsicum annuum ) subjected to chilling temperatures were assessed. Application of the growth regulator paclobutrazol affected plant growth and fruit morphology. The plants were more compact and the fruits were less elongated than control fruits. There was about 60% more plasma membrane on a fresh weight basis from treated fruits. At harvest there was no difference in sterol to phospholipid ratio, or in phospholipid fatty acid composition of control compared with paclobutrazol treated fruit. However, plasma membrane ATPase acitivity of treated fruit was two times higher than that of control fruit. After storage at chilling temperature (2°C), the control fruit developed more chilling iniury, and had greater weight loss and a higher rate of K⁺ leakage than paclobulrazol treated fruit. Plasma membrane phospholipid content decreased and saturation of phospholipid fatty acids was higher than in control fruit. These two changes were largely absent in plasma membrane from treated fruit. At harvest antioxidant levels in the plasma membrane of paclobutrazol treated peppers were higher than in those of controls and changed little during storage, whereas levels in control fruit plasma membrane decreased 66%. ATPase activity increased and then decreased in control fruit held at low temperature, whereas in treated fruit activity was constant. The protective effect of paclobutrazol against chilling injury of pepper fruit may result from a combination of its effect on fruit morphology, and protection of the lipids against oxidative stress.     

The effect of acclimation temperature on the activation energies of state III respiration and on the unsaturation of membrane lipids of goldfish mitochondria.

The influence of the acclimation temperature on the thermotropic behaviour of mitochondrial respiration and on the degree of unsaturation of mitochondrial membrane lipids has been studied. The mitochondria were isolated from red muscle, white muscle and liver of goldfish acclimated to 5, 20 and 30 degrees C. ADP-activated succinate oxidation was measured at different temperatures and resulted in non-linear Arrhenius-plots with breaks between 10 and 23 degrees C. As for the break-temperatures, there was found a shift downwards in preparations of decreased acclimation temperatures. This could be caused by a changed composition of membrane lipids and a simultaneous shift of the membrane phase transition temperature. Therefore, the fatty acid composition of all membrane preparations was analyzed. However, no consistent change of the degree of unsaturation due to a changed acclimation temperature could be found.     

采后荔枝果实冷害过程中多胺含量的变化

以桂味荔枝果实为材料,研究冷害及外源亚精胺(Spd)处理对果实膜透性和内源多胺含量变化的影响。结果表明,在0℃下贮藏时果实发生冷害过程中,荔枝果皮中腐胺(Put)、Spd、精胺(Spm)含量在14d后明显增加,膜透性快速增大,21d时果皮出现明显的冷害褐变,Put进一步积累,而Spm含量下降,Spd保持较高水平。非冷害温度(3℃)下贮藏时,果皮多胺含量变化相对较小。0℃下果肉的多胺含量和变化幅度低于3℃果实,并延迟7d衰老。外源Spd处理明显提高果实内源多胺含量的同时,延缓了果皮相对膜透性增加,减轻了冷害。这表明果皮中Put的积累可能是荔枝果实冷害的结果,冷藏初期Spm含量的上升可能是果实对冷害的防卫反应。     

Propofol Prevents Oxidative Stress by Decreasing the Ischemic Accumulation of Succinate in Focal Cerebral Ischemia–Reperfusion Injury

Oxidative stress caused by mitochondrial dysfunction during reperfusion is a key pathogenic mechanism in cerebral ischemia–reperfusion (IR) injury. Propofol (2,6-diisopropylphenol) has been proven to attenuate mitochondrial dysfunction and reperfusion injury. The current study reveals that propofol decreases oxidative stress injury by preventing succinate accumulation in focal cerebral IR injury. We evaluated whether propofol could attenuate ischemic accumulation of succinate in transient middle cerebral artery occlusion in vivo. By isolating mitochondria from cortical tissue, we also examined the in vitro effects of propofol on succinate dehydrogenase (SDH) activity and various mitochondrial bioenergetic parameters related to oxidative stress injury, such as the production of reactive oxidative species, membrane potential, Ca²⁺-induced mitochondrial swelling, and morphology via electron microscopy. Propofol significantly decreased the ischemic accumulation of succinate by inhibiting SDH activity and inhibited the oxidation of succinate in mitochondria. Propofol can decrease membrane potential in normal mitochondria but not in ischemic mitochondria. Propofol prevents Ca²⁺-induced mitochondrial swelling and ultrastructural changes to mitochondria. The protective effect of propofol appears to act, at least in part, by limiting oxidative stress injury by preventing the ischemic accumulation of succinate.