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.

Transperineal ultrasound in the assessment of haemorrhoids and haemorrhoidectomy: a pilot study.

BACKGROUND: The purpose of the study was the measurement of the anal cushion area using static transperineal ultrasound in a group of patients with symptomatic grade III and IV haemorrhoids about to undergo haemorrhoidectomy and compare them with a group of age-matched normals and the measured area following haemorrhoidectomy. METHODS: Transperineal sonography was performed using a linear transducer measuring the anal cushion area by subtracting the measured luminal diameter of the undisturbed anal canal from the inner border of the internal anal sphincter. Measures were made 6 weeks following haemorrhoidectomy. RESULTS: Comparisons were made between 22 normals and 36 patients with haemorrhoids (31 evaluable post-operatively). The median area of normals was 0.78 cm², that of pre-operative patients 2.25 cm² and that of post-operative cases 1.20 cm². There was a significant difference between pre- and post-operative cases with cushion areas of normal patients being significantly lower than post-operative cases. Variance of measurement in all 3 groups was negligible. CONCLUSION: Static transperineal sonography measuring the anal cushion area is reproducible and shows marked differences between normals and patients with symptomatic haemorrhoids. There is a marked effect on measured area resultant from haemorrhoidectomy.

Analysis of collagen fibre shape changes in breast cancer.

Second harmonic generation microscopy was performed on both normal and diseased breast tissue. Differences in the collagen fibre shape between normal, benign and malignant breast tissue were compared and quantified using elliptical Fourier analysis. Principal shape analysis of these coefficients provided an understanding of the key differences in collagen fibre shape between the three tissue types. A Gaussian model was also used to associate the shape of the fibre with the probability that it had been sampled from malignant breast tissue. These results provide quantitative evidence for the alteration of collagen fibre shape in both benign and malignant breast tissue.

Myelin structure is a key difference in the x-ray scattering signature between meningioma, schwannoma and glioblastoma multiforme.

Small angle x-ray scattering (SAXS) patterns of benign and malignant brain tumour tissue were examined. Independent component analysis was used to find a feature set representing the images collected. A set of coefficients was then used to describe each image, which allowed the use of the statistical technique of flexible discriminant analysis to discover a hidden order in the data set. The key difference was found to be in the intensity and spectral content of the second and fourth order myelin scattering peaks. This has clearly demonstrated that significant differences in the structure of myelin exist in the highly malignant glioblastoma multiforme as opposed to the benign: meningioma and schwannoma.

Wavelet-based feature extraction applied to small-angle x-ray scattering patterns from breast tissue: a tool for differentiating between tissue types.

This paper reports on the application of wavelet decomposition to small-angle x-ray scattering (SAXS) patterns from human breast tissue produced by a synchrotron source. The pixel intensities of SAXS patterns of normal, benign and malignant tissue types were transformed into wavelet coefficients. Statistical analysis found significant differences between the wavelet coefficients describing the patterns produced by different tissue types. These differences were then correlated with position in the image and have been linked to the supra-molecular structural changes that occur in breast tissue in the presence of disease. Specifically, results indicate that there are significant differences between healthy and diseased tissues in the wavelet coefficients that describe the peaks produced by the axial d-spacing of collagen. These differences suggest that a useful classification tool could be based upon the spectral information within the axial peaks.

Implication of NOTCH1 gene in susceptibility to anxiety and depression among sexual abuse victims.

Sexual abuse contributes to the development of multiple forms of psychopathology, including anxiety and depression, but the extent to which genetics contributes to these disorders among sexual abuse victims remains unclear. In this translational study, we first examined gene expression in the brains of rodents exposed to different early-life conditions (long, brief or no maternal separation). Hypothesizing that genes revealing changes in expression may have relevance for psychiatric symptoms later in life, we examined possible association of those genes with symptoms of anxiety and depression in a human sample of sexual abuse victims. Changes in rodent brain gene expression were evaluated by means of correspondence and significance analyses of microarrays by comparing brains of rodents exposed to different early-life conditions. Tag single-nucleotide polymorphisms (SNPs) of resulting candidate genes were genotyped and tested for their association with symptoms of anxiety and depression (Hospital Anxiety and Depression Scale) in a sample of 361 sexual abuse victims, using multinomial logistic regression. False discovery rate was applied to account for multiple testing in the genetic association study, with q-value of 0.05 accepted as significant. We identified four genes showing differential expression among animals subjected to different early-life conditions as well as having potential relevance to neural development or disorders: Notch1, Gabrr1, Plk5 and Zfp644. In the human sample, significant associations were observed for two NOTCH1 tag SNPs: rs11145770 (OR=2.21, q=0.043) and rs3013302 (OR=2.15, q=0.043). Our overall findings provide preliminary evidence that NOTCH1 may be implicated in the susceptibility to anxiety and depression among sexual abuse victims. The study also underscores the potential importance of animal models for future studies on the health consequences of early-life stress and the mechanisms underlying increased risk for psychiatric disorders.

Stress ulceration in rats: impact of prior stress experience.

The role of prior stress experience on the response of the organism to a later stressor has been studied in terms of the extent of gastric ulceration induced by the second stressor. These studies have focused largely on effects of prior stress (shock, restraint, activity) on the ulceration developing under later restraint and activity stress. The studies indicate that prior exposure to restraint stress provides some protection against later restraint ulcer development. Using shock as the prior stressor, the effects on later restraint stress are determined by the particular characteristics of the shock (controllability and predictability), and by contextual factors. Studies are lacking on the significance of these characteristics of shock for later activity stress ulcers. Further studies are required therefore on the significance of psychological characteristics of the prestress, and on the effects of these prestressors at different stages of the life cycle. The identification of these factors, and a clearer picture of the protective and exacerbating effect of prior stress will allow us to explore the physiological (central and peripheral) mechanisms underlying ulcer development and ulcer susceptibility.

Restraint stress in biomedical research: an update.

Since the publication of our initial review of restraint stress in 1986, much work has continued with this technique, either as a tool for the investigation of other pharmacological, physiological, or pathologic phenomena or with restraint stress itself serving as the object of the study. As we noted in 1986, the major use of restraint has been for the induction of stress responses in animals and, more specifically, for the investigation of drug effects, particularly as they affect typical stress-related pathology--gastrointestinal, neuroendocrine, and immunological agents have been extensively studied. In compiling this update on restraint stress and its effects, we noted an increasing emphasis on central nervous system mechanisms in peripheral disease, especially gastrointestinal disease. In particular, many CNS-active agents have been tested for their effects on gastric and duodenal lesion formation and gastric secretion, including antidepressants, antipsychotics, anxiolytics, noradrenergic, serotonergic, dopaminergic, and peptidergic compounds. Some of these agents are especially active in the gastrointestinal tract even when administered centrally, further solidifying the concept of a brain-gut axis. The present update includes studies of: methods and procedures, pre-restraint manipulations, post-restraint/healing effects, and drug effects. In addition, a current bibliography of reports that have employed restraint is included.

Hypothalamo-pituitary-adrenal manipulations and stress ulcerations in rats.

Both gastric ulceration and activation of the hypothalamo-pituitary adrenal axis are considered integral to the stress response, and a causal relationship between the two has been suggested. In the present study, corticosterone secretion in rats was either stimulated with CRF or lowered with metyrapone during a known ulcerogenic stress. Reduction of circulating corticosterone during the stress had no effect on ulceration severity compared to saline-treated stressed control rats. Treatment with CRF in stressed animals reduced ulceration severity. The mechanism of this protective effect remains unclear. The findings do not support a simple causal relationship between adrenocortical activity and gastric ulceration.

Poststress effects of danger and safety signals on gastric ulceration in rats.

Gastric ulceration of rats stressed by restraint in 19 degrees C water for 75 min was markedly increased by allowing a 75-min postrestraint room-temperature rest period during which the rat was exposed to cues that had previously been associated with the delivery of 80 5-s uncontrollable electric shocks distributed over four sessions. This effect obtained equally without regard to whether the "danger cues" were punctate signals or constant contextual cues or whether contextual ones were interrupted by punctate safety signals. The experimental treatments used were unusual in that they equated the groups on their total conditioning history and thus allowed a more pure look at the poststress effect than heretofore. Other groups provided controls for prior shocks, rest, and their interaction as well as handling. Analyses of corticosterone after the stress or stress-rest cycle revealed only a general decline in corticosterone levels with rest undifferentiated across groups.

Juvenile and adult footshock stress modulate later adult gastric pathophysiological reactions to restraint stresses in rats.

Rats were stressed with (a) signaled footshocks, (b) unsignaled footshocks, or (c) handled without footshocks as a prepubertal juvenile (28-36 days old), as an adult (96 days old), or both. This yielded nine treatment groups (3 x 3). Two days after the adult treatment, all animals were challenged by restraint and partial immersion in water (19 degrees C) to assess their relative susceptibility to gastric erosions ("ulcers"). We found that any prior exposure to footshock stress increased the amount of ulcers; juvenile and adult experiences each produced equal increases but the combination of the two was less ulcerogenic than either alone. The predictability of the footshocks did not modulate ulcerogenicity. Adult corticosterone responses to (a) adult stress and (b) ulcer induction were not related to the observed ulcer severity; however, juvenile footshock stress appeared to reduce the corticoid response to the ulcerogenic challenge but not to the adult footshock stress.

Quality of poststressor rest influences the ulcerative process.

Gastric ulceration in rats is exacerbated by allowing a so-called recovery period after exposure to an ulcerogenic stressor. One hypothesis, which has support from pharmacological studies, argues that this effect is brought about by a rebound of parasympathetic activation. We tested this parasympathetic rebound hypothesis by presenting animals with a fear-inducing (sympathetic-activating) conditioned stimulus (CS) after 2 hr of water-restraint stress. Contrary to the hypothesis, presentation of such a CS increased severity of ulceration compared with those animals that did not receive the CS after restraint stress and control animals. These ulceration data favor instead a sustained activation hypothesis for ulceration, whereby presentation of the CS effectively prolonged the length of time during which animals were under stress, thus enhancing the degree of ulceration. Measurement of plasma corticosterone however indicated a negative correlation between adrenocortical activity and degree of gastric ulceration, contrary to that expected by a sustained activation hypothesis. It is suggested that this inconsistency may be because of the activating of a pituitary-endorphinogenic mediated stress analgesia.

Amino Acid Effects on Post-Stress Ulcers: Relationship to Brain Serotonin, 5-HIAA and Norepinephrine.

To replicate and extend results of earlier studies on amino acid effects on post-stress ulcers, rats were subjected to i.p. injections of (a) saline, (b) tryptophan, (c) tyrosine + valine or(d) tryptophan + tyrosine + valine, either 30 minutes before or immediately after one hour of water restraint stress. Gastric lesions, brain norepinephrine, serotonin and 5-hydroxyindoleacetic acid (5-HIAA) were examined after one hour of poststress rest. We hypothesised that post-stress lesions could be aggravated by central noradrenergic hypoactivity and serotonergic hyperactivity during the post-stress period. Other studies have indicated that tyrosine + valine reduces central serotonergic activity, while additional tryptophan blocks this effect. We therefore expected post-stress lesions to be reduced in tyrosine + valine but not in tryptophan + tyrosine + valine treated animals. Although these expectations were met tentatively in animals injected prior to stress, thus replicating tyrosine + valine effects we had observed earlier, opposite results were found in animals treated post-stress. The brain analyses indicate that the data cannot be explained by a norepinephrine/serotonin imbalance hypothesis. The time dependency of the effects underlines the need for caution in clinical applications of these amino acid treatments.

Parallelism among stress effects on ulcer, immunosuppression and analgesia: commonality of mechanisms?

Some general principles and mechanisms have been discovered that govern proactive effects of one stress on animals' later stress or challenge induced behavior, nociception, immune function, and stomach ulcerations. As described below, these principles demonstrate a considerable degree of parallelism across the domains represented by measures of learning deficits and fear-related behaviors, hypoalgesia, immunological status and stress gastric ulcerations. At a minimum, these parallels suggest that operational factors found important in one domain are likely factors of importance in the other domains. Beyond this, these parallels are permissive of an inference of commonality in underlying processes. Nonetheless, results reviewed below suggest that the specific response of the organism to stress is highly dependent on specific characteristics of the stressor.

Intracerebroventricular neuropeptide Y suppresses open field and home cage activity in the rat.

Effects of intracerebroventricular administration of neuropeptide Y on open field behaviour, behavioural habituation and corticosterone response to open field testing, and on home cage activity have been investigated in the rat. In the open field, NPY reduced activity in a dose-dependent manner. Behavioural habituation was not influenced. After 5 days of recovery, NPY-treated animals did not differ from non-treated in any of the measured parameters. Peripheral corticosterone levels were not significantly affected, although there was a strong tendency towards an increase. Injection of 2 nmol NPY did not produce any gross neurological deficits. At this dose, NPY greatly suppressed home cage activity. The effect lasted throughout the recording period of 22 h, abolishing the normal circadian variation in activity. After 5 days of recovery, the effect was no longer present. Our interpretation of these findings is, that NPY is a highly potent endogenous agent capable of producing certain important aspects of behavioural sedation in a reversible manner. Since NPY did not decrease the corticosterone response to a novel stimulus, its pattern of actions seems to differ from synthetic sedative drugs.

Anxiety and helplessness in the face of stress predisposes, precipitates, and sustains gastric ulceration.

When an organism is subjected to stress, gastric ulcers or ulcerations commonly develop but the vulnerability to and amount of pathology induced varies considerably between individuals. The role of psychological factors in determining the occurrence and severity of these ulcerations is amply demonstrated in the studies reviewed here. The present paper (a) gives a brief history of the search for the causes of gastric ulcer, (b) provides a review of our own research which reveals that vulnerability to gastric ulceration is modulated by psychologically meaningful experiences, and (c) offers a multifactorial perspective on the causes of gastric ulceration and the future of research on it.

The effects of lesions of the superior colliculus on locomotor orientation and the orienting reflex in the rat.

The effects of bilateral removal of the superior colliculus or visual cortex on visually guided locomotor movements in rats performing a brightness discrimination task were investigated directly with the use of cine film. Rats with collicular lesions showed patterns of locomotion comparable to or more efficient than those of normal animals when approaching one of 5 small doors located at one end of a large open area. In contrast, animals with large but incomplete lesions of visual cortex were distinctly impaired in their visual control of approach responses to the same stimuli. On the other hand, rats with collicular damage showed no orienting reflex or evidence of distraction in the same task when novel visual or auditory stimuli were presented. However, both normal and visual-decorticate rats showed various components of the orienting reflex and disturbance in task performance when the same novel stimuli were presented. These results suggest that although the superior colliculus does not appear to be essential to the visual control of locomotor orientation, this midbrain structure might participate in the mediation of shifts in visual fixation and attention. Visual cortex, while contributing to visuospatial guidance of locomotor movements, might not play a significant role in the control and integration of the orienting reflex.

Effect of central noradrenaline depletion on corticosterone levels and gastric ulcerations in rats.

Effects of central noradrenergic depletion on the stress responses of rats were explored using the new selective neurotoxin (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4)). Noradrenergic depletion using DSP-4 was followed by a reduction in basal corticosterone levels after 7 days. Three weeks after DSP-4 treatment, animals exhibited less severe and fewer gastric ulcerations than control animals following 23 h immobilization stress, but stress levels of corticosterone were similar for the two groups. No differences could be found in the peripheral gastric levels of noradrenaline between experimental and control animals, while central noradrenaline was reduced to approximately 30% of control levels. The data support previous findings using other methods that central noradrenaline is an important factor in stress-induced gastric ulceration. The peripheral mechanisms for the protective effects of DSP-4 remain to be elucidated, and studies of these may cast light on the efferent pathways between the central nervous system and gastric mucosa which are involved in stress-induced gastric pathology.

Intracerebroventricular neuropeptide Y protects against stress-induced gastric erosion in the rat.

The effects of intracerebroventricular administration of neuropeptide Y (NPY) on stress-induced gastric erosion in the rat were investigated. Animals were exposed to water immersion stress after a single injection of 2 nmol NPY. Stress-induced erosion was reduced by approximately 50%. The plasma corticosterone levels were not affected. We interpret the protective effect of NPY as a manifestation of its sedative properties.

The effect of sleep deprivation and recovery sleep on plasma corticosterone in the rat.

The effect of 24-h sleep deprivation by forced locomotion on plasma corticosterone was investigated in the rat. Corticosterone was slightly elevated after 21.5 h sleep deprivation, but did not differ from controls after a 2.5-h recovery period. An acute 20-min forced locomotion period caused a marked rise in plasma corticosterone. It is concluded that stress is not a major factor contributing to the massive effects of sleep deprivation on sleep parameters.