Strain and gender specific effects in the forced swim test: effects of previous stress exposure

The chronic mild stress (CMS) procedure was developed in rodents to target anhedonia, the core symptom of depressive melancholia. Stress exposure has been shown to induce a variety of physiological, biochemical and behavioral alterations associated with depression, although its anhedonic consequences as indexed by either sucrose intake and preference or thresholds for brain stimulation reward are less reliably observed. In the present study, we assessed the effects of six weeks of CMS on the latter measure in two strains of male and female rats subsequently challenged with an acute psychophysical stressor, forced swimming; their behavior in the swimming cylinder was evaluated on two consecutive days. While brain stimulation reward thresholds and response rates were unchanged by CMS exposure, significant differences in forced swim behaviors were observed between male control and CMS groups. In particular, male Long Evans rats with a history of CMS showed the largest decrease in the duration of active behaviors on the second test day, a pattern less evident in the Sprague-Dawley strain of rats, or in any of the female groups. The results suggest that the effects of depressogenic manipulations are strain and gender dependent, with male Long Evans rats most susceptible, as demonstrated by the selective reduction of struggling behaviors. Inclusion of multiple measures, including the forced swim test, would provide a better understanding of the psychopathological profile engendered by chronic exposure to mild stressors and its genetic specificity.     

Permanent and transient effects of repeated preweaning stress on social and sexual behaviors of rats

We have reported that exposure of preweaning male and female rats to a model of unpredictable mild physical stressors (Neo-A) can decrease the behavioral and hormonal responses to acute and chronic stress as adults. In this paper we have analyzed the effect of Neo-A on development of social behaviors, including aggressiveness, social dominance, and sexual behavior in adulthood. The subjects were divided into two groups: Neo-A (daily exposed to unpredictable mild stresses- from day 2 up to day 15 of suckling; n = 30 litters) and controls (C) (undisturbed rats, except for testing, during the same period of life; n = 26 litters). When day 6 pups were submitted to a social clustering test the Neo-A group showed a higher rate of litter-mate clustering than C. The 35 days Neo-A males and Neo-A females submitted to a social behavior test after 24 h of social isolation also showed higher scores of time spent in active social interaction than controls, as well as a higher ratio of animals showing aggressive playing. A second social behavior test performed after 48 h of social isolation at days 75-80 of age revealed that only Neo-A females displayed increased social behavior and aggressive behaviors, whereas controls did not. A water competition test performed at 24 and 48 h after water deprivation showed that Neo-A adult males spent more time in possession of the drinking device and drank more frequently than C. When adult proestrous females were exposed to a sexual behavior test, the Neo-A group showed shorter latency and higher scores of lordosis quotient. These results support the view that exposure to this model of repeated mild stress early in life stimulates the development of social behavior, dominance and sexual behavior.     

Chronic stress, but not hypercaloric diet, impairs vascular function in rats

The aim of this study was to evaluate vascular and metabolic effects of chronic mild unpredictable stress (CMS) and hypercaloric diet (HD) without carbohydrate supplementation in rats. Male Sprague-Dawley rats were randomly assigned to four groups: Control, HD, CMS, and HD plus CMS. CMS consisted of the application of different stressors for 3 weeks. The rats were killed 15 days after CMS exposure. The HD group presented higher plasma lipid concentrations, without changes in fasting glucose concentration, glucose tolerance test, and vascular function and morphology, in comparison with the control group. Stressed rats presented higher fasting blood concentration of insulin, higher homeostasis model assessment index values and area under the curve in an oral glucose tolerance test, in comparison with non-stressed rats. CMS increased the plasma concentrations of corticosterone and lipids, and the atherogenic index values, without change in high-density lipoprotein level. CMS increased intima-media thickness and induced endothelium-dependent supersensitivity to phenylephrine, and lowered the relaxation response to acetylcholine in the thoracic aorta isolated from rats fed with control or HD, in comparison with non-stressed groups. CMS effects were independent of diet. In non-stressed rats, the HD induced dyslipidemia, but did not change glucose metabolism, vascular function, or morphology. The data from this study indicate that CMS promotes a set of events which together can contribute to impair function of the thoracic aorta.     

Circadian activity of the hypothalamic-pituitary-adrenal axis is differentially affected in the rat chronic mild stress model of depression

The altered activity of the hypothalamic-pituitary-adrenal (HPA) axis is often observed in stress-related disorders. According to the literature, about 60% of patients with major depressive disorder elicit high levels of cortisol. It is still unclear why high cortisol levels are not observed in all patients. In this study, we used the chronic mild stress (CMS) rat model of depression, which is based on continuous exposure to unpredictable stressors, to track longitudinal changes in HPA function using fecal corticosterone metabolites (FCM) as a read out. The dexamethasone suppression test was used to assess negative feedback inhibition of the HPA axis. Our results show (1) a disturbance in diurnal corticosterone rhythm measured as fluctuations of the diurnal FCM peak, (2) differences in corticosterone levels between stress-susceptible and stress-resilient animals, (3) recovery of diurnal corticosterone rhythm after 8 weeks of CMS, and (4) alterations in sensitivity to dexamethasone in negative feedback regulation of corticosterone secretion during the time course of CMS. Thus, a disruption of HPA axis circadian rhythmicity coincides with the initial state in the development of depression-like behavior. This chronobiological abnormality, as well as the hypersecretion of corticosterone, is state, rather than trait, dependent.     

Sexual experience and preference for males or females in the female rat

The influence of sexual experience on preference behavior was investigated in adult female rats. In the first experiment, preference behavior for sexually active males versus estrous females was investigated in female rats who were experienced with mounting behavior. Preference of these mount experienced females was compared to preference of females who were naive in this respect. Mount experience with estrous females induced a female-directed preference in ovariectomized female rats, irrespective of whether the females were treated with OIL or with testosterone-propionate (500 micrograms TP, injected once 48 h prior to testing). Sexually naive, OIL-treated females did not show a preference for males or females, but TP induced a preference for a male. Individual differences in mount frequency were not correlated with preference behavior. In the second experiment, the influence of experience with feminine sexual responses on preference behavior was investigated. OIL-treated and TP-treated females oriented equally toward males and females when they had been given the opportunity to copulate with males prior to the preference tests. The results strongly suggest that prior sexual experience is an important determinant of preference behavior. The female's behavior during sexual interactions was, however, not predictive of later preference.     

Predictors of the first onset of a major depressive episode and changes in depressive symptoms across adolescence: stress and negative cognitions

This 6-year longitudinal study examined stressors (e.g., interpersonal, achievement), negative cognitions (self-worth, attributions), and their interactions in the prediction of (a) the first onset of a major depressive episode (MDE), and (b) changes in depressive symptoms in adolescents who varied in risk for depression. The sample included 240 adolescents who were first evaluated in Grade 6 (M = 11.86 years old; SD = 0.57; 54.2% female) and then again annually through Grade 12. Stressful life events and depressive diagnoses were assessed with interviews; negative cognitions and depressive symptoms were assessed with self-report questionnaires. Discrete time hazard modeling revealed a significant interaction between interpersonal stressors and negative cognitions, indicating that first onset of an MDE was predicted by high negative cognitions in the context of low interpersonal stress, and by high levels of interpersonal stressors at both high and low levels of negative cognitions. Analyses of achievement stressors indicated significant main effects of stress, negative cognitions, and risk in the prediction of an MDE, but no interactions. With regard to the prediction of depressive symptoms, multilevel modeling revealed a significant interaction between interpersonal stressors and negative cognitions such that among adolescents with more negative cognitions, higher levels of interpersonal stress predicted higher levels of depressive symptoms, whereas at low levels of negative cognitions, the relation between interpersonal stressors and depression was not significant. Risk (i.e., maternal depression history) and sex did not further moderate these interactions. Implications for intervention are discussed.     

Enduring deficits in contextual and auditory fear conditioning after adolescent, not adult, social instability stress in male rats

Adolescence is a time of developmental changes and reorganization in the brain and stress systems, thus, adolescents may be more vulnerable than adults to the effects of chronic mild stressors. Most studies, however, have not directly compared stress experienced in adolescence to the same stress experience in adulthood. In the present study, adolescent (n=46) and adult (n=48) male rats underwent 16 days of social instability stress (daily 1h isolation and change of cage partners) or were non-stress controls. Rats were then tested on the strength of acquired contextual and cued fear conditioning, as well as extinction learning, beginning either the day after the stress procedure or 3 weeks later. No difference was found among the groups during the Training Phase of conditioning. Irrespective of the time between the social stress experience and fear conditioning, rats stressed in adolescence had decreased context and cue memory, and cue generalization compared to control rats, as measured by the percentage of time spent freezing in tests. Social instability stress in adulthood had no effect on any measure of fear conditioning. The results support the hypothesis that adolescence is a time of heightened vulnerability to stressors.     

应激性抑郁样行为发生中海马5-羟色胺1A受体的作用及其对NMDA受体和AMPA受体的调节

为探讨慢性不可预见性温和应激(chronic unpredictable mild stress, CUMS)诱发抑郁样行为发生中海马5-羟色胺1A受体(5-hydroxytryptamine receptor 1A, 5-HT1AR)表达与作用, 及其对谷氨酸N-甲基-D-天冬氨酸(N-methyl-D-aspartic acid, NMDA)受体和α-氨基羟甲基异恶唑丙酸(α-amino-3-hydroxy-5- methylisoxazole-4-propionic acid, AMPA)受体的影响。通过建立CUMS动物模型, 给应激抑郁模型大鼠海马微量注射5-HT1A受体激动剂、给正常大鼠海马微量注射5-HT1A受体拮抗剂, 测量大鼠体重变化率, 并采用糖水偏爱测试、旷场实验和悬尾实验等方法对大鼠进行行为学检测, 运用Western blot和ELISA方法检测大鼠海马组织中5-HT1AR和NMDAR和AMPAR的关键亚基的表达以及磷酸化水平。结果显示, 与对照组相比, CUMS组大鼠表现出抑郁样行为, 海马5-HT1AR、AMPA受体的GluR2/3亚基表达及磷酸化明显降低, NMDA受体的NR1和NR2B亚基表达及磷酸化显著增加; 正常大鼠海马微量注射5-HT1A受体拮抗剂WAY100635, 动物行为学表现及AMPA受体、NMDA受体表达及磷酸化水平均与CUMS组相同; 注射5-HT1A受体激动剂8-OH-DPAT能逆转应激诱导的上述改变。以上结果表明, CUMS诱发抑郁样行为与海马5-HT1AR表达下降, AMPAR表达量及磷酸化水平降低, NMDAR表达量及磷酸化水平升高有关。5-HT通过5-HT1AR产生抗抑郁作用。5-HT1AR激动剂抗抑郁作用与降低NMDAR表达量及磷酸化水平, 提高AMPAR表达量及磷酸化水平密切相关。     

Persistence of flavor neophobia as an indicator of state-dependent retention induced by pentobarbital, stress, and estrus

As fear conditioning paradigms have failed to find state-dependent retention (SDR) in rats dissociated by the estrous state, an attempt was made to establish a paradigm involving the attenuation of flavor neophobia as a more sensitive indicator of SDR. Experiment 1 established that the paradigm did indicate SDR in male rats dissociated by pentobarbital (10 mg/kg). Experiment 2 generalized the utility of the technique by employing an endogenous state, shock-induced stress, as the dissociative state. Having proved to be an indicator of SDR in the first two experiments, the technique was used in Experiment 3 to investigate the possible dissociative characteristics of the estrous state in female rats. Estrus was investigated because, due to the hormonal condition during estrus, it may provide a useful tool in investigating the dissociative aspects of human perimenstrual periodic psychosis. In Experiment 3, using ovariectomized female rats brought into estrus by injection of estrogen and progesterone in a sequence and amount comparable to the natural cycle, estrus was not demonstrated to be dissociative, i.e., did not produce SDR. The implications of these results in relation to perimenstrual periodic psychosis were discussed.     

Effects of stress on defensive aggression and dominance in a water competition test

Water-deprived rats in a pair competing for a single source of water quickly establish a firm relationship during which one rat drinks consistently more (dominant) than the other (submissive) animal. This relationship is formed during the first competition and is very stable during subsequent tests. Exposure of dominant rats to a severe (18 hrs immobilization), but not a mild (2 hrs immobilization), stressor reduced markedly aggressive behavior and inverted transiently the dominant submissive relationship of the pairs. Exposure of submissive rats to the severe stressor resulted in only minor reductions of aggressive behavior in these animals. Prestress anxiety predicted stress effects in the dominant animals in that high-anxious animals lost more dominant behavior and weight during stress as compared with low-anxious rats. Thus, severe stress can transiently reduce dominant but not submissive behavior during water competition and high-anxious rats are more prone to lose their aggressive behavior.     

Gender differences in acute and chronic stress in Wistar Kyoto (WKY) rats.

While females are considered more susceptible to depressive behavior, this assertion is not strongly supported by the experimental literature. Since stress contributes to depressive behavior, male and female Wistar Kyoto (WKY) rats were exposed to either one session (acute stress) or 5 sessions (chronic stress) of restraint plus cold in order to study depressive behavior in male and female rats. After their respective treatment exposure, rats were tested in the open field test (OFT) and for retention of a passive-avoidance (P-A) task. One stress session resulted in significant immobility in the OFT for males, whereas 5 sessions were required to produce similar immobility in female rats. Acute stress interfered with the retention of the P-A response for males, while both acute and chronic stress produced poor P-A responses in female rats. Food consumption decreased progressively, as a function of stress sessions, in female rats, whereas feeding in males returned to control levels after five stress days. Both acute and chronic stress exacerbated the stress ulcer response in male rats, but not in female rats. Chronic, but not acute, stress resulted in an increase in serotonin transporter mRNA levels in the dorsal raphe nucleus of both male and female rats. The general consensus from these data suggested that female rats were more vulnerable to chronic stress and consequently supported the notion that females may be more susceptible to stress-induced behavioral depression. Key Words: WKY rats, acute and chronic stress, gender, passive avoidance, open field behavior, stress-ulcer, adrenal weight, serotonin, dorsal raphe nucleus     

The Effectiveness of Strategies for Increasing Social Interaction with a Physically Disabled Person1

The effectiveness of strategies a physically disabled person can use to increase a nondisabled person's willingness to engage in social interaction were investigated. In the strategies, the disabled person engages in behavior demonstrating that he/she is not preoccupied with the disability. Female college students chose the amount of time to be spent listening to music versus social interaction before and after learning that the other was a disabled male. The subject witnessed via “closed circuit” TV (actually a videotape) a brief interaction between the male and another female in the waiting room from which the subject had just come. In the Interest in others condition, the male encouraged the female to talk about herself. In the Typical activities condition, the male mentioned he had to pick up tickets, organize a party, and study for an exam. In the Athletic activities condition, the male mentioned he went swimming and bowling. In the Request condition, the male asked the female to pick up a paper he had dropped. In the Control condition, there was limited interaction. In the Inappropriate communication condition, the male mentioned the same activities as in the Typical activities condition but talked continuously without waiting for polite interest from the female. Changes in preference for social interaction were significantly more positive in the Attentive to others, Typical activities and Athletic activities conditions than in the Control conditions. It was concluded that demonstrating interest in others, typical activities and athletic activities are effective strategies.     

Changes in Behaviour and Body Weight Following a Single or Double Social Defeat in Rats

In a series of experiments, the consequences of a single and double social conflict on various behaviours and body weight in rats were studied. Animals were subjected to social defeat by placing them into the territory of an aggressive male conspecific for one hour, either once, or twice at the same time on two consecutive days. To assess the consequences of social defeat, three experiments were performed with independent groups of rats. In the first experiment, an open field test was performed two days after the last conflict. Locomotor activity was strongly reduced after social defeat. There were no differences between the single and double defeat group. To assess the effects of social defeat on subsequent social behaviour, a second experiment was performed in which experimental animals were confronted with an unfamiliar non-aggressive rat two days after a single or double conflict. Social defeat resulted in a reduction of social contact with the unfamiliar conspecific. There was no difference between the single and double conflict group. In the third experiment, the effects of social conflict on food intake, body weight and saccharine preference were measured. Food intake was not affected after a single conflict, but in the double conflict group food intake was decreased for several days. Body weight gain was decreased after both single and double social defeat. The decrease was stronger in the double conflict group. Water intake and saccharine preference were not significantly affected. This study revealed that social defeat in rats causes pronounced changes in various behaviours and body weight. Different aspects of behaviour are differentially affected by defeat with respect to the magnitude and time course of the changes induced. Moreover, different behavioural parameters are differentially sensitive to repetition of the stressor.     

Influences of stress and situation-specific mastery beliefs and satisfaction with social support on well-being and academic performance.

The present study examined the effects of stress and cumulative situation-specific mastery beliefs and satisfaction with social support on somatic and psychological symptomatology and academic performance of 146 college men. Stress was directly related to increased symptomatology and decreased grade point average, whereas mastery beliefs were directly associated with reduced depression and anxiety. Satisfaction with social support was related to anxiety through an interaction with mastery. Neither mastery nor satisfaction with support appeared to act as a stress-buffer through a statistical interaction with stress to moderate effects of high stress. However, earlier analysis of some of the same data showed that total stress, the best predictor of each outcome in this study, was reduced by the combination of strong situation-specific mastery beliefs and high satisfaction with support for subjects who faced many stressors. The combined findings show that situation-specific mastery beliefs reduced appraisal of stress and were associated with better psychological adjustment.     

The role of anxiety sensitivity in subjective and physiological responses to social and physical stressors

This study examined the specificity of the relationship between anxiety sensitivity (AS), a measure of catastrophizing about arousal-related sensations, and pain responses, by examining the effect of AS on responses to stressors of a physical and social nature. Healthy men and women (n = 129) between the ages of 18 and 25 years were recruited from the community to participate in a study examining subjective, cognitive and behavioural responses to different types of stressors. Participants were randomly assigned to one of 3 groups: (i) a neutral condition in which they sat quietly and read a popular magazine; (ii) a social stress condition in which they anticipated having to give a self-disclosing speech; and (iii) a physical stress condition in which they were presented with 3 countdown to shock trials where a mild electrical shock was administered on the non-dominant arm. Subjective ratings and physiological responses were recorded in anticipation of the stressor and immediately after stress exposure. Results indicated that AS was indirectly related to pain ratings via its effect on anticipatory anxiety ratings. AS was associated with anticipatory anxiety ratings, regardless of whether the stressor was of a physical or social nature. Furthermore, AS was not shown to be directly associated with exaggerated subjective or physiological reactions to the physical stressor. These results indicate that the role of AS in pain responses may be mediated through a global effect on anxiety, and limited to the anticipatory stage of the pain experience. If future studies yield similar findings in pain patients, then they would suggest that interventions for helping individuals high in AS should focus on catastrophic thinking in anticipation of stressors in general, rather than on pain-specific stressors.     

Long-term effects of footshock and social defeat on anxiety-like behaviours in rats: Relationships to pre-stressor plasma corticosterone concentration

We compared the consequences of two stressors, 'unnatural' inescapable footshocks (IFSs) and 'natural' social defeat (SD), on behaviours typically sensitive to stress [sucrose preference, open field (OF), elevated plus maze (EPM) and acoustic startle responses (ASRs)] and the association with pre-stressor plasma corticosterone concentration. After initial blood sampling, rats (n?=?20 per group) were exposed to either 10 IFSs (1?mA intensity, 5?s duration each) or to 1?h SD (defeat by an aggressive resident male rat and further exposure but separated in a small cage) or to control procedures (handling). Rats were tested once for ASR (day 19), while the other behavioural tests were applied once weekly for 3 weeks. Both stress groups showed short-lasting lowered sucrose preference, and in the EPM they showed shorter total distance moved, shorter distance moved on open arms and less time on open arms compared to controls. In the OF test, IFS rats showed shorter total distance moved up to 2 weeks after stress. The SD group showed shorter total distance moved in the OF, which was only significant 2 weeks after stress. Low pre-stressor plasma corticosterone concentration was only associated with defecation (IFS rats) and latency to enter open arms in the EPM (all low corticosterone subgroups, n?=?10 per subgroup). SD rats with high initial plasma corticosterone concentration showed enhanced ASR compared to the other subgroups with high initial plasma corticosterone concentration (n?=?9 per subgroup). The results indicate that footshock and SD, while generally leading to an increase in anxiety behaviours, represent qualitatively different stressors.     

Responsiveness to estrous chemostimuli in male rats (Rattus norvegicus) of different ages

Previous findings indicate that Norway rats (Rattus norvegicus) respond differentially to estrous chemostimuli in early infancy and that adult male sexual responsiveness to such chemostimuli depends on infantile experience with suckling odors (Fillion & Blass, in press). The development of behavioral responsiveness to estrous chemostimuli in male rats from late infancy to adulthood was examined in this study. Preweanling, adolescent, and adult males were observed for 20 min in the presence of an anesthetized estrous or an anesthetized diestrous female. This was, for adolescent and adult subjects, the first heterosexual encounter since weaning. Rats in each age-group responded differentially according to female state, but in distinct, age-related ways. Preweanling spent more time investigating the female's body if she was estrous. Adults expressed the same affinity but by more focused investigation directed perivaginally. Adolescents showed no female-directed bias but groomed themselves more if the female was estrous. These data identify developmental continuities and discontinuities in male rat behavior toward sexually receptive females.     

The Effect of Selective Serotonin Releasing Agents in the Chronic Mild Stress Model of Depression in Rats

Chronic exposure to mild unpredictable stress has been found to depress the consumption of, and preference for, highly palatable sucrose solution in rats. Stress-induced behavioral deficits may be maintained for a long time, however chronic administration of clinically effective antidepressants can restore normal behavior. This is the first report showing that Sprague-Dawley rats can be used in this model. A preference deficit in this strain of rats took at least 7 weeks to develop; about twice the time required when hooded Lister or Wistar rats are used in this model. Water consumption was not effected by chronic exposure to the mild stress regime and/or by chronic administration of the selective serotonin (5-HT) releasing agent MMAI (5-methoxy-6-methyl-2-aminoindan). The stress-induced deficit in sucrose intake was completely reversed by chronic treatment with MMAI (5 mg/kg, 2 x day) over 3 weeks in the two-bottle tests. In single-bottle tests, chronic treatment with the selective 5-HT releasers, MMAI (5 mg/kg, 2 x day) or MTA (p-methylthioamphetamine; 5 mg/kg, 2 x day), reversed the deficit in rewarded behavior (anhedonia) measured as a decrease in the consumption of 1% sucrose solution in the chronic mild stress model of depression in rats. With the experimental procedure employed, and at a dose of 10 mg/kg/day of 5-HT releasers, the magnitude and onset of this effect were greater than observed following similar administration of the selective 5-HT reuptake inhibitor (SSRI) sertraline (10 mg/kg/day), used as a standard anti-depressant drug.     

Intrasexual interactions among female golden hamsters (Mesocricetus auratus) over the estrous cycle

Social interactions of cycling female golden hamsters paired with ovariectomized animals in large enclosures were primarily agonistic over the 4-day estrous cycle. These aggressive interactions were intense as indicated by frequent occurrences of chase and flight behavior. Dominant and subordinate social ranks were established in the majority of pairs (96%) tested, and even females in sexual heat were capable of attacking and dominating their rivals. Furthermore, cycling females exhibited significantly more aggressive acts than ovariectomized opponents 1 day prior to sexual receptivity. No differences in fighting patterns were found between animals on the other 3 days of the estrous cycle. Additional analyses revealed clear differences in agonistic elements, including flank marking, between dominant and subordinate females regardless of whether they were gonadectomized or intact. These analyses also showed that dominant individuals frequently chased and bit opponents during encounters within their nesting area. The data are examined for implications on the adaptive organization of female hamster agonistic behavior and the neuroendocrine regulation of species-typical behavior over the estrous cycle.     

Burnout‐depression overlap: Nomological network examination and factor‐analytic approach

Burnout has been defined as a condition in which individuals are left exhausted by a long‐term confrontation with unmanageable job stressors. The question of whether burnout reflects anything other than depressive responses to unresolvable stress remains an object of debate. In this 911‐participant study (83% female; mean age: 42.36), we further addressed the issue of burnout‐depression overlap. Burnout was assessed with the exhaustion subscale of the Maslach Burnout Inventory‐General Survey (MBI ‐GS ) and depression with the PHQ ‐8. The relationships of burnout and depression with three job‐related variables – illegitimate work tasks, work‐nonwork interference, and job satisfaction – and three “context‐free” variables – social support, general health status, and trait anxiety – were examined. Burnout and depression were found to be strongly correlated, to cluster together, and to exhibit overlapping nomological networks. Remarkably, the average correlations of burnout and depression with job‐related variables were almost identical. A principal component analysis and a principal axis factor analysis both showed that the items of the MBI ‐GS and of the PHQ ‐8 loaded on a single dimension. All in all, our findings are consistent with the view that burnout is a depressive condition. The distinction between burnout and depression may be an instance of the jangle fallacy.