Adipokinetic hormone-induced enhancement of antioxidant capacity of Pyrrhocoris apterus hemolymph in response to oxidative stress

The in vivo effects of oxidative stress on adipokinetic hormone (AKH) titer in short-winged (brachypterous) males of the firebug Pyrrhocoris apterus were tested using paraquat (PQ), a bipyridilium herbicide. PQ undergoes a cyclic redox reaction with oxygen during microsomal and electron transfer reactions forming free radicals in the insect body. Oxidative insult (40 pmol PQ) resulted in enhanced protein carbonylation (a biomarker for oxidative stress) and a depletion of glutathione (GSH) pool in the hemolymph. Interestingly, AKH titer was significantly enhanced in hemolymph at 4 h post inoculation of PQ, while its content in CNS (brain with corpora cardiaca) showed non-specific changes in comparable period. Co-injection of AKH with PQ (40 pmol each) reversed these effects by decreasing protein carbonyl formation, increasing reduced GSH levels, and enhancing the total antioxidant capacity of cell free plasma. Our results indicate that there is a positive feedback regulation between an oxidative stressor action and the level of AKH in insect body, and that AKHs might be involved in the activation of antioxidant protection mechanism.     

Individual behaviour in firebugs (Pyrrhocoris apterus)

The concept of animal personalities has recently become of major interest as researchers began to wonder why animals within a given population show consistent behaviour across situations and contexts, what led to the evolution of such behavioural inflexibility and what mechanisms might underlie the phenomenon. A recent model explains individual differences in a population as the result of trade-off between present and future reproduction. We tested this model on the two wing morphs, i.e. short-winged (brachypterous) and long-winged (macropterous) specimens of the firebug (Pyrrhocoris apterus). Since it has been already demonstrated that the two wing morphs differ in their life-history strategies, this species is an ideal subject to test whether the specimens with different life-history strategies have different personalities as well. The results show that individuals behave consistently over time and across contexts, meaning observed bugs do have personalities. We also have found that in females, the two wing morphs have different personalities supporting the theoretical predictions, i.e. winged ones, which are supposed to have lower future reproductive value, are braver and more exploratory. We found no difference between the morphs in males. Differences in reproductive investment might explain this discrepancy between the sexes.     

Induction of adult diapause in Pyrrhocoris apterus L. by short cold exposure

Summary The ripening of ovaries in the females of Pyrrhocoris apterus can be inhibited not only by short-day photoperiod but also by cold shocks during the first week of the adult life.Ovaries in a part of the females are prevented from ripening by transferring the bugs from 25° C and long-day photoperiod to substantially lower temperatures for 2 (or 4) days. The oviposition is inhibited at least for 3 weeks after the return to original conditions favourable for reproduction. This effect is achieved when cold treatment is combined either with darkness or with long day. A difference between 2 and 4 days of chilling was not recorded. The effect of cold exposure (evaluated in our experiments by the proportion of mature and immature females) depends on the temperature of the cold exposure and on the age of females at the time of transfer to cold. The incidence of immature females was higher when lower temperatures (+2 to +6° C) were used and when females were transferred to cold early (0–2 days) after the adult moult.     

Termination of adult diapause in Pyrrhocoris apterus (Heteroptera: Pyrrhocoridae) in the field

The time when diapause ends in the Czech population of Pyrrhocoris apterus was studied by the transfer of the overwintering bugs to laboratory conditions of 25°C and long day (L 18, D 6) or short day (L 12, D 12).Both the incidence of oviposition after the transfer to short-day conditions and the duration of the pre-oviposition period after the transfer to long-day conditions indicate that the diapause ends in the period of December–January. Development is then inhibited environmentally, by low temperatures.Thus the diapause of P. apterus is terminated before the advent of hard frosts. The resistance to frost is not bound to the condition of the intrinsic inhibition of the maturation of gonads.

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Zusammenfassung Die Termination der Diapause kann nicht — wie vielfach angenommen wird — mit der Wiederaufnahme der Entwicklung gleichgesetzt werden, da in der Natur die Entwicklung nach dem Ende der Diapause meist durch niedrige Temperaturen noch weiterhin inhibiert bleibt (Quieszenz). Um das wahre Ende der Diapause bei Pyrrhocoris apterus zu ermitteln, wurden Wanzen im Verlaufe der Überwinterung (Hiberntion) (August–März) in abgestuften Intervallen aus dem Freien ins Laboratorium übertragen, wo 25° und 24-stündige Photoperioden mit Photophasen von 12 oder 18 std herrschten.Die Versuche wurden während drei Winterperioden durchgeführt (1964–65, 1966–67, 1967–68) und dabei Wanzen aus drei klimatisch unterschiedlichen Lokalitäten: Smolenice, Slaný und Jankovice (Seehöhe 240, 280 bzw. 430 m) verglichen, deren mittlere Jahres-temperatur 9,0, 8,2 bzw. 7.5° beträgt. Die Wanzen wurden paarweise bei Lindensamen und Wasser gehalten. In allen Versuchen wurde Eiablage, bei manchen auch Kopulation, Eianzahl und Schlüpfrate, täglich ermittelt.Im Langtag (L 18, D 6) begannen die Weibchen stets Eier zu legen, doch verkürzte sich im Verlaufe der Überwinterung schrittweise die Länge der Präovipositionsperiode. Zwischen August und Anfang Oktober begannen die Eiablagen in der Regel nach 26–34 Tagen. Nach Mitte Januar (manchmal auch schon ab Dezember) verkúrzte sich die Präovipositionsperiode auf nur 8–9 Tage, was dem Wert der Präovipositionszeit bei Nondiapause-Tieren entspricht.Im Kurztag (L 12, D 12) blieben die diapausierenden Wanzen unfruchtbar. Die Weibchen der August- und September-Proben legten überhaupt keine Eier (obwohl sie zwei über drei Monate lang gehalten wurden. Die Weibchen der im Oktober und November eingeholten Proben legten nur zu einem geringen Prozentsatz (14 bzw. 32%) und nach längerer Präovipositionsperiode ab. Bereits ab Mitte Dezember legten jedoch alle Weibchen nach einer Präovipositionsperiode von nur 8–11 Tagen ab.In der Natur endet also die imaginale Diapause bei tschechischen Populationen von P. apterus schon im Dezember, gegebenenfalls im Januar, d.h. noch vor Eintritt strengerer Fröste. Der Adaptationswert der Diapause beruht daher nicht auf einer direkten Erhöhung der Frostresistenz, sondern auf der Synchronisation des Lebenszyklus der Insekten mit der Jahreszeit.Die photoperiodische Termination der imaginalen Diapause bei P. apterus wird mit ähn-lichen Erscheinungen bei anderen Insektenarten verglichen.

     

Antioxidant S-allylcysteine prevents gentamicin-induced oxidative stress and renal damage

Acute renal failure (ARF) is a major complication of gentamicin (GM) treatment, which is effective against gram-negative infections. Since experimental evidence suggests a role of reactive oxygen species (ROS) in GM-induced ARF, in this work we studied the effect of a garlic-derived compound, S-allylcysteine (SAC), which is a free radical scavenger, on GM-induced nephrotoxicity. In rats treated with GM (70 mg/kg/12 h/4 days/s.c.), ARF was evident by the: (i) decrease in creatinine clearance and increase in blood urea nitrogen, (ii) decrease in blood glutathione peroxidase (GPx) activity and increase in urinary excretion of N-acetyl-beta-D-glucosaminidase and total protein, and (iii) necrosis of proximal tubular cells. These alterations were prevented by SAC treatment (250 mg/kg/i.p. 24 h before the first dose of GM and 125 mg/kg/12 h/4 days along GM-treatment). Furthermore, SAC prevented the GM-induced oxidative stress (protein carbonyl groups) and the decrease in manganese superoxide dismutase (Mn-SOD), GPx, and glutathione reductase (GR) activities in renal cortex. In conclusion, SAC ameliorates the GM-induced ARF by a mechanism related, at least in part, to its ability to decrease oxidative stress and to preserve antioxidant enzymes activity in renal cortex.     

Regulation of body size in a heteropteran bug, Pyrrhocoris apterus

The determination of body size (length or weight) of teneral adults of Pyrrhocoris apterus L. was investigated. About 50% of size variation was determined already in the egg stage (average egg weight adult length correlations were 0.67 in males and 0.71 in females). During the larval development, starvation, low temperature, and pollution by excreta combined with crowding reduced adult size and usually increased the development length. The effects were directly proportionate to the length of the stress period, and increased with larval age. Photoperiod or intrinsic variation of development time did not affect adult size. The effects of stress were largely compensated when it was followed by a period of compensation growth at optimum conditions. A dramatic reduction of size appeared when last inster larvae starved and moulted to adults without access to food. Average adult weight was then reduced by up to 48%. Threshold weight for adult moulting was 18 mg, being attained after 1–2 day feeding of the last instar.

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Zusammenfassung Die Körpergrösse bei den Imagines eines Wildstammes der Feuerwanze Pyrrhocoris apterus wurde teilweise durch die Veränderlichkeit der Eigrösse determiniert, teilweise auch durch Hungern der Larven, niedrige Temperatur und metabolischen Produkten beeinflusst. Der Effekt letzterer Faktoren stieg mit deren Dauer und war je grösser desto näher sich die Wirkungsperiode zur Imaginalhäutung befand. Unterschiedliche Photoperioden wirkten auf die Körperlänge nicht. Wurden die Larven zwischen der ungünstigen Periode und der Imaginalhäutung noch den optimalen Bedingungen ausgesetzt, kompensierten sich die negativen Einflüsse weitgehend. Zu einer drastischen Verkleinerung der Imagines kam es nur, wenn das letzte Larven-stadium mit Hunger beeinflusst wurde und bis zur Imaginalhäutung keinen Zutritt zum Futter hatte. Minimales Gewicht, wo die Metamorphose der Larven noch möglich war, wurde zu 18 mg festgestellt.

     

Function of the neuroendocrine complex in diapausing Pyrrhocoris apterus females

The role of the brain in inhibiting the action of corpora allata in diapausing short-day females was investigated by transplantation experiments. The function of the transplanted glands was evaluated by oviposition. Active glands from long-day females remained active for a long period of time after transplantation into short-day females, although in situ corpora allata were inhibited shortly after the transfer of females from long to short day. Moreover, inactive glands from short-day females became active after transplantation into other short-day females. In contrast, corpora allata remained inhibited when transplanted together with the brain in the neuroendocrine complex (brain-corpora cardiaca-corpus allatum) where the nervous connections between the brain and corpus allatum remained intact. It is therefore suggested that short-day conditions inhibit corpora allata via nervous connections with the brain.     

Enzymatic capacity for accumulation of polyol cryoprotectants changes during diapause development in the adult red firebug, Pyrrhocoris apterus

Abstract. Diapausing adults of the red firebug, Pyrrhocoris apterus, were maintained in the laboratory at constant conditions of 20 °C and short days (LD 12 : 12 h) for 5 months. Spontaneous termination of diapause is seen in 0.4% of adults. At different times of diapause development, groups of adults were exposed to an 8‐week gradual cold treatment ending at 0 °C. Ribitol and sorbitol contents remain very low at constant 20 °C and their rapid accumulation only occurrs at temperatures below 5 °C. The capacity to accumulate ribitol in response to low temperature stimulus remains relatively stable but the capacity to accumulate sorbitol decreases to zero during the 5 months of diapause development. Glycogen (whole‐body and fat‐body reserves) accumulates relatively rapidly during the early phase (up to 1–2 months) and slowly depletes during the late phase of diapause development under constant conditions. Upon cold treatment, part of the glycogen reserve was depleted. The activities of most enzymes involved in polyol metabolism (namely glycogen phosphorylase, glucose‐6‐P dehydrogenase, phosphofructokinase, aldose reductase, polyol dehydrogenase and ketose reductase) increase relatively rapidly during the early phase of diapause development and, in the late phase, they either become stable or slowly decrease. Cold treatment has either no effect or results in a moderate increase in activity when applied in the early phase of diapause development but results in a more or less obvious decrease of enzymatic activity when applied in the late phase.     

Locomotor activity in adult Pyrrhocoris apterus (Heteroptera) in relation to sex, physiological status and wing dimorphism

The pattern of locomotor (walking) activity was studied in adult males and females of short‐winged (brachypterous) and long‐winged (macropterous) morph of the flightless bug Pyrrhocoris apterus (Linnaeus) (Heteroptera: Pyrrhocoridae) under constant laboratory conditions. Walking activity was measured with a computerized video system and analysed with respect to sex, physiological status (reproduction, diapause and reproductive arrest of non‐diapause type) and wing dimorphism of the bugs. The largest duration was observed in the macropterous females with reproductive arrest of non‐diapause type (average 6 h per day) and the shortest duration in diapausing brachypterous females and males (average less than 2 h per day). This was reflected also in the overall time spent by walking during the first 14 days after imaginal ecdysis. The time spent by walking significantly increased in the macropterous morph as the bugs aged, whereas in diapausing brachypters the time spent by walking decreased with age. No linear relationship between walking activity and age was found in reproductive brachypterous morph. The bugs of all experimental groups moved mostly during the photophase and were almost inactive during the scotophase. Thus, walking activity in P. apterus is diurnal, irrespective of the wing morph, physiological status, sex and age. Contrary to the macropterous morph, where the locomotor activity of females during photophase was significantly higher than in males, no significant differences were found between the locomotor activities of brachypterous males and females. The observed differences in locomotor activity are discussed in relation to different roles of two wing morphs in the life history of this heteropteran.     

The effect of allatectomy and photoperiod on the supercooling point in Pyrrhocoris apterus adults

Abstract Adults of Pyrrhocoris apterus (L.) (Heteroptera: Pyrrhocoridae) reared at 26°C had a considerably lower supercooling point (SCP) in diapause-inducing (LD 12:12 h) than in diapause-inhibiting (LD 18:6 h) photoperiodic regimes. Exposure of the insects to diapause-inducing acclimation conditions (LD 8:16 h and a temperature of 20°C during photophase and 5°C during scotophase) for 2–3 weeks had very little effect on the SCP, irrespective of the rearing photoperiod. Allatectomized adults kept continuously at LD 18:6 h and 26°C had high SCP similar to intact or sham-operated insects. In contrast to non-allatectomized insects, the SCP of allatectomized insects decreased after an exposure of from 2 to 3 weeks to diapause-inducing acclimation conditions (see above) almost to the level found in the intact diapausing insects. The relationship between the decrease of SCP and ‘diapause syndrome’ (de Wilde, 1970) is discussed.     

Selection for an unstable micropterism in Pyrrhocoris apterus (Heteroptera: Pyrrhocoridae)

Twenty-eight generations of mass selection for an unstable micropterism (short wings) in the red firebug, Pyrrhocoris apterus (L.), resulted in an unstable micropterous ( ump ) strain with a high frequency of micropterous individuals. The strain, segregating individuals with enormously reduced wings, is polymorphic for mosaic pigmentation in the epidermis. Micropterous individuals, both symmetric and asymmetric, appear predominantly among pigmentation mosaic ( ump _mo) forms. Individuals with a normal, nonmosaic epidermis ( ump _wt) are rarely micropterous. The reproductive capacity of micropterous ump _mo females is strongly reduced. It is suggested that pigmentation mosaicism and unstable micropterism may have a common or linked genetic basis. A new mode of wing length regulation is suggested, engaged in conspicuous destabilization of ump wings and consisting presumably of transpositions of mobile genetic elements.     

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.     

Stevioside prevents oxidative stress in wheat seedlings

This is the first study on the effect of stevioside, a diterpene glycoside that is a new promising plant growth regulator, on the antioxidant and photosynthetic systems of seedlings of the winter wheat cultivar Kazanskaya 560. Stevioside has been demonstrated to cause a decrease in the malondialdehyde formation rate, an increase in the activities of antioxidant enzymes (peroxidase and ascorbate peroxidase), and the accumulation of proline and carotenoids. Apparently, this integrated effect of stevioside can prevent oxidative stress caused by adverse environmental factors in plants.     

Behavioral regulation of developmental cycle in Pyrrhocoris apterus L. (Heteroptera: Pyrrhocoridae)

Summary During the warm season the larvae of Pyrrhocoris apterus select in their natural habitats places with temperatures which do not exceed 24°C on average even at the hours of maximum sunshine. At the same time other places of their habitats have temperatures up to 50°C. This active behavioral selection of the microclimate regulates the speed of P. apterus development and entertains the monovoltine cycle in climatic, conditions of Central Europe. A less rigorous choice of the microclimate, even when higher temperatures were tolerated and the development was not disturbed by them, would cause the disturbance of the monovoltine cycle and would probably endanger the survival of the population.     

Carboxyfullerene prevents iron-induced oxidative stress in rat brain

Carboxyfullerene, a water-soluble carboxylic acid derivative of a fullerene, was investigated as a protective agent against iron-induced oxidative stress in the nigrostriatal dopaminergic system of anesthetized rats. Intranigral infusion of exclusive carboxyfullerene did not increase lipid peroxidation in substantia nigra or deplete dopamine content in striatum. Infusion of ferrous citrate (iron II) induced degeneration of the nigrostriatal dopaminergic system. An increase in lipid peroxidation in substantia nigra as well as decreases in K+-evoked dopamine overflow and dopamine content in striatum were observed 7 days after the infusion. Co-infusion of carboxyfullerene prevented iron-induced oxidative injury. Furthermore, tyrosine hydroxylase-immunoreactive staining showed that carboxyfullerene inhibited the iron-induced loss of the dopaminergic nerve terminals in striatum. The antioxidative action of carboxyfullerene was verified by in vitro studies. Incubation of brain homogenates increased the formation of the Schiff base fluorescent products of malonaldehyde, an indicator of lipid peroxidation. Both autooxidation (without exogenous iron) and iron-induced elevation of lipid peroxidation of brain homogenates were suppressed by carboxyfullerene in a dose-dependent manner. Our results suggest that intranigral infusion of carboxyfullerene appears to be nontoxic to the nigrostriatal dopaminergic system. Furthermore, the potent antioxidative action of carboxyfullerene protects the nigrostriatal dopaminergic system from iron-induced oxidative injury.     

Vitamin C prevents cigarette smoke-induced oxidative damage in vivo

Our recent in vitro results [4] indicate that cigarette smoke induces oxidation of human plasma proteins and extensive oxidative degradation of the guinea pig lung, heart, and liver microsomal proteins, which is almost completely prevented by ascorbic acid. In this paper, we substantiate the in vitro results with in vivo observations. We demonstrate that exposure of subclinical or marginal vitamin C-deficient guinea pigs to cigarette smoke causes oxidation of plasma proteins as well as extensive oxidative degradation of the lung microsomal proteins. Cigarette smoke exposure also results in some discernible damage of the heart microsomal proteins. The oxidative damage has been manifested by SDS-PAGE, accumulation of carbonyl and bityrosine, as well as loss of tryptophan and protein thiols. Cigarette smoke exposure also induces peroxidation of microsomal lipids as evidenced by the formation of conjugated dienes, malondialdehyde, and fluorescent pigment. Cigarette smoke-induced oxidative damage of proteins and peroxidation of lipids are accompanied by marked drop in the tissue ascorbate levels. Protein damage and lipid peroxidation are also observed in cigarette smoke-exposed pair-fed guinea pigs receiving 5 mg vitamin C/animal/day. However, complete protection against protein damage and lipid peroxidation occurs when the guinea pigs are fed 15 mg vitamin C/animal/day. Also, the cigarette smoke-induced oxidative damage of proteins and lipid is reversed after discontinuation of cigarette smoke exposure accompanied by ascorbate therapy. The results, if extrapolated to humans, indicate that comparatively large doses of vitamin C may protect the smokers from cigarette smoke-induced oxidative damage and associated degenerative diseases.     

Garlic supplementation prevents oxidative DNA damage in essential hypertension

Oxygen-free radicals and other oxygen/nitrogen species are constantly generated in the human body. Most are intercepted by antioxidant defences and perform useful metabolic roles, whereas others escape to damage biomolecules like DNA, lipids and proteins. Garlic has been shown to contain antioxidant phytochemicals that prevent oxidative damage. These include unique water-soluble organosulphur compounds, lipid-soluble organosulphur compounds and flavonoids. Therefore, in the present study, we have tried to explore the antioxidant effect of garlic supplementation on oxidative stress-induced DNA damage, nitric oxide (NO) and superoxide generation and on the total antioxidant status (TAS) in patients of essential hypertension (EH). Twenty patients of EH as diagnosed by JNC VI criteria (Group I) and 20 age and sex-matched normotensive controls (Group II) were enrolled in the study. Both groups were given garlic pearls (GP) in a dose of 250 mg per day for 2 months. Baseline samples were taken at the start of the study, i.e. 0 day, and thereafter 2 months follow-up. 8-Hydroxy-2′-deoxyguanosine (8-OHdG), lipids, lipid peroxidation (MDA), NO and antioxidant vitamins A, E and C were determined. A moderate decline in blood pressure (BP) and a significant reduction in 8-OHdG, NO levels and lipid peroxidation were observed in Group I subjects with GP supplementation. Further, a significant increase in vitamin levels and TAS was also observed in this group as compared to the control subjects. These findings point out the beneficial effects of garlic supplementation in reducing blood pressure and counteracting oxidative stress, and thereby, offering cardioprotection in essential hypertensives.     

Melatonin administration prevents lipopolysaccharide-induced oxidative damage in phenobarbital-treated animals

The protective effect of melatonin on lipopolysaccharide (LPS)-induced oxidative damage in phenobarbital-treated rats was measured using the following parameters: changes in total glutathione (tGSH) concentration, levels of oxidized glutathione (GSSG), the activity of the antioxidant enzyme glutathione peroxidase (GSH-PX) in both brain and liver, and the content of cytochrome P450 reductase in liver. Melatonin was injected intraperitoneally (ip, 4mg/kg BW) every hour for 4 h after LPS administration; control animals received 4 injections of diluent. LPS was given (ip, 4 mg/kg) 6 h before the animals were killed. Prior to the LPS injection, animals were pretreated with phenobarbital (PB), a stimulator of cytochrome P450 reductase, at a dose 80 mg/kg BW ip for 3 consecutive days. One group of animals received LPS together with N^w-nitro-L-arginine methyl ester (L-NAME), a blocker of nitric oxide synthase (NOS) (for 4 days given in drinking water at a concentration of 50 mM). In liver, PB, in all groups, increased significantly both the concentration of tGSH and the activity of GSH-PX. When the animals were injected with LPS the levels of tGSH and GSSG were significantly higher compared with other groups while melatonin and L-NAME significantly enhanced tGSH when compared with that in the LPS-treated rats. Melatonin alone reduced GSSG levels and enhanced the activity of GSH-PX in LPS-treated animals. Additionally, LPS diminished the content of cytochrome P450 reductase with this effect being largely prevented by L-NAME administration. Melatonin did not change the content of P450 either in PB- or LPS-treated animals. In brain, melatonin and L-NAME increased both tGSH levels and the activity of GSH-PX in LPS-treated animals. The results suggest that melatonin protects against LPS-induced oxidative toxicity in PB-treated animals in both liver and brain, and the findings are consistent with previously published observations related to the antioxidant activity of the pineal hormone.