Tamoxifen and pregnancy: an absolute contraindication?

PURPOSE: Breast cancer is the most common malignancy among young women of reproductive age. Adjuvant treatment with tamoxifen reduces the risk of recurrence in hormone-sensitive breast cancer. However, the use of tamoxifen is considered contraindicated during pregnancy, because of a limited number of case reports demonstrating potential adverse effects on the fetus. The objective of this report is to give a more broad overview of the available data on the effect of tamoxifen exposure during pregnancy. METHODS: A literature review was performed using PubMed and the databases of the Netherlands Pharmacovigilance Centre Lareb and of the International Network on Cancer, Infertility, and Pregnancy. RESULTS: A total of 238 cases of tamoxifen use during pregnancy were found. Of the 167 pregnancies with known outcome, 21 were complicated by an abnormal fetal development. The malformations described were non-specific and the majority of cases concerned healthy infants despite exposure to tamoxifen. CONCLUSION: There seems to be an increased risk of fetal abnormalities when taking tamoxifen during pregnancy (12.6% in contrast to 3.9% in the general population), but the evidence is limited and no causal relationship could be established. The possible disadvantage of postponing or discontinuing tamoxifen for the maternal prognosis is unclear. Patients should be counseled about the use of tamoxifen during pregnancy instead of presenting it as being absolutely contraindicated.

Emergence of Escherichia coli encoding Shiga toxin 2f in human Shiga toxin-producing E. coli (STEC) infections in the Netherlands, January 2008 to December 2011.

The Shiga toxins of Shiga toxin-producing Escherichia coli (STEC) can be divided into Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2) with several sub-variants. Variant Stx2f is one of the latest described, but has been rarely associated with symptomatic human infections. In the enhanced STEC surveillance in the Netherlands, 198 STEC O157 cases and 351 STEC non-O157 cases, including 87 stx2f STEC isolates, were reported between 2008 and 2011. Most stx2f strains belonged to the serogroups O63:H6 (n=47, 54%), O113:H6 (n=12, 14%) and O125:H6 (n=12, 14%). Of the 87 stx2f isolates, 84 (97%) harboured the E. coli attaching and effacing (eae) gene, but not the enterohaemorrhagic E. coli haemolysin (hly) gene. stx2f STEC infections show milder symptoms and a less severe clinical course than STEC O157 infections. Almost all infections with stx2f (n=83, 95%) occurred between June and December, compared to 170/198 (86%) of STEC O157 and 173/264 (66%) of other STEC non-O157. stx2f STEC infections in the Netherlands are more common than anticipated, and form a distinct group within STEC with regard to virulence genes and the relatively mild disease.

Recurrent long-lasting tethering reduces BDNF protein levels in the dorsal hippocampus and frontal cortex in pigs.

Brain-derived neurotrophic factor (BDNF) signaling has been implicated in the onset of depression and in antidepressant efficacy, although the exact role of this neurotrophin in the pathophysiology of depression remains to be elucidated. Also, the interaction between chronic stress, which may precede depression, corticosteroids and BDNF is not fully understood. The present study aimed at investigating whether long-lasting, recurrent tethering of sows during a period of 1.5 or 4.5 years leads to enduring effects on measures that may be indicative of chronic stress, compared with animals kept in a group housing system ('loose' sows). Immediately after slaughter, the frontal cortex, dorsal and ventral hippocampus were dissected and protein levels of BDNF and its receptors were analyzed and compared with plasma cortisol levels and adrenal weights. Results indicate that tethering stress reduced BDNF protein levels in the dorsal hippocampus and the frontal cortex, but not in the ventral hippocampus. In addition, levels of TrkB, the high affinity receptor for BDNF, were increased in the dorsal hippocampus. Plasma cortisol levels and adrenal weight were increased after tethering. These stress effects on BDNF levels were more pronounced after 4.5 years of recurrent tethering and negatively correlated in particular in the frontal cortex with cortisol levels and adrenal weight. This suggests that the stress effect of tethered housing on neurotrophin levels may be mediated via cortisol. Taken together, these data indicate that recurrent tethering stress in sows over 4.5 years results in a loss of neurotrophic support by BDNF, mediated by an overactive neuroendocrine system.

Systematic lymphadenectomy in patients with clinical stage II endometrial carcinoma: a case report and review of the literature.

OBJECTIVE: The aim of this case report and review of the literature was to evaluate the effect of adding pelvic and/or para-aortic lymphadenectomy to hysterectomy and bilateral salpingo-oophorectomy (BSO) on the five year recurrence-free survival in patients with clinical Stage II endometrial carcinoma. MATERIALS AND METHODS: A Pubmed, Embase, and Cochrane library search was performed to identify relevant articles. After screening, using predetermined exclusion and inclusion criteria, and critical appraisal, a final of four articles remained. RESULTS: This search only revealed studies with a retrospective design. Two articles showed a significant disease-specific survival benefit in patients undergoing systematic lymphadenectomy for Stage II endometrial carcinoma. In multivariate analyses, conducted in both studies, this improvement in survival was also evident (HR 0.75, 95% CI 0.69 - 0.81, p < 0.001 and HR 0.74, 95% CI 0.58 - 0.93, p = 0.0096). The remaining studies revealed a non-significant ten-year recurrence-free survival (77% vs 65%) and five-year overall survival (72% vs 70%) in favour of patients undergoing systematic lymphadenectomy. CONCLUSION: The practise of performing a systematic lymphadenectomy in patients with clinical Stage II endometrial carcinoma as advocated in guidelines, is not based on evidence from randomised clinical trials. However, lymph node dissection seems to improve the five-year disease-specific survival in retrospective studies.

Prevalence, characterisation and clinical profiles of Shiga toxin-producing Escherichia coli in The Netherlands.

Detection of Shiga toxin-producing Escherichia coli (STEC) in The Netherlands is traditionally limited to serogroup O157. To assess the relative importance of STEC, including non-O157 serogroups, stool samples submitted nationwide for investigation of enteric pathogens or diarrhoea were screened with real-time PCR for the presence of the Shiga toxin genes. Patients were selected if their stool contained blood upon macroscopic examination, if they had a history of bloody diarrhoea, were diagnosed with haemolytic uraemic syndrome, or were aged <6 years (irrespective of the bloody aspect of the stool). PCR-positive stools were forwarded to a central laboratory for STEC isolation and typing. In total, 4069 stools were examined, with 68 (1.7%) positive PCR results. The highest prevalence was for stools containing macroscopic blood (3.5%), followed by stools from patients with a history of bloody diarrhoea (2.4%). Among young children, the prevalence (1.0%) was not significantly higher than among random, non-bloody, stool samples from diarrhoeal patients (1.4%). STEC strains were isolated from 25 (38%) PCR-positive stools. Eleven O-serogroups were detected, including five STEC O157 strains. As serogroup O157 represented only 20% of the STEC isolates, laboratories should be encouraged to use techniques enabling them to detect non-O157 serogroups, in parallel with culture, for isolation and subsequent characterisation of STEC strains for public health surveillance and detection of outbreaks.

Repeated social defeat in female pigs does not induce neuroendocrine symptoms of depression, but behavioral adaptation.

The aim of this study was to develop an animal model of major depression. Since two thirds of depressive patients are women, it is important to develop specific female animal models of depression. We therefore determined the consequences of chronic social defeat in individually housed prepubertal female pigs confronted with a dominant, older pig. Repeated defeat increased the salivary cortisol level, measured immediately after the confrontations, but this effect diminished after repeated confrontations. Neither organ weights nor the number of glucocorticoid (GR) and mineralocorticoid (MR) receptors in the ventral hippocampus were affected by repeated defeat. Serotonin turnover in the dorsal hippocampus was also unaffected. Behavioral analysis revealed that across confrontations, the pigs reduced the time spent actively attacking the dominant pigs, whereas the time increased in which the pigs passively underwent aggression and/or actively avoided aggression. Therefore, we conclude that the repeated social defeat paradigm does not induce long-lasting depression-like neuroendocrine effects as a consequence of behavioral adaptations (changes in the fighting strategy) in the young female pigs.

Comparison of microscopy, real-time PCR and a rapid immunoassay for the detection of Giardia lamblia in human stool specimens.

Giardia lamblia is one of the most common intestinal parasites worldwide, with microscopy being the diagnostic reference standard for use with human stools. However, microscopy is time-consuming, labour-intensive and lacks sensitivity when single stools are examined. In the present study, microscopy, real-time PCR and a rapid immunoassay were compared for the detection of G. lamblia in human stools. All three methods were highly sensitive, with values of 99%, 100% and 98%, respectively. Specificity and positive and negative predictive values were >or=97%, except when using real-time PCR, for which the specificity and positive predictive value were 92% and 93%, respectively. The lower specificity of real-time PCR was associated mostly with failure to detect specimens regarded as true positives for G. lamblia DNA, although cross-contamination was suspected in a minority of cases because of the large amount of G. lamblia DNA present in most positive specimens. It was concluded that microscopy should remain the primary diagnostic tool for identifying G. lamblia in human stools, mainly because of its ability to simultaneously detect other gastrointestinal parasites. However, the simple and rapid immunoassay is a valuable tool to decrease turn-around time. Real-time PCR provides additional sensitivity, although there is a risk of cross-contamination. Based on this observation, and the need for other real-time assays to be developed to detect other intestinal parasites, real-time PCR is currently useful only as an additional test supplementary to microscopy.

Detection of diarrhoea-causing protozoa in general practice patients in The Netherlands by multiplex real-time PCR.

The diagnostic value of a multiplex real-time PCR for the detection of Entamoeba histolytica, Giardia lamblia and Cryptosporidium parvum/Cryptosporidium hominis was evaluated by comparing the PCR results obtained with those of routinely performed microscopy of faecal samples from patients consulting their general practitioner (GP) because of gastrointestinal complaints. Analysis of 722 faecal DNA samples revealed that the prevalence of G. lamblia was 9.3% according to PCR, as compared to 5.7% by microscopy. The number of infections detected was more than double in children of school age. Furthermore, G. lamblia infection was detected in 15 (6.6%) of 228 faecal samples submitted to the laboratory for bacterial culture only. C. parvum/C. hominis infections were not diagnosed by routine procedures, but DNA from these organisms was detected in 4.3% of 950 DNA samples. A strong association with age was noted, with Cryptosporidium being detected in 21.8% of 110 children aged <5 years. C. hominis was the most prevalent species. E. histolytica was not detected in this study population. Analysis of microscopy data revealed that the number of additional parasites missed by PCR was small. Overall, the study demonstrated that a multiplex real-time PCR approach is a feasible diagnostic alternative in the clinical laboratory for the detection of parasitic infections in patients consulting GPs because of gastrointestinal symptoms.

Long-term vitamin D deficiency in older adult C57BL/6 mice does not affect bone structure, remodeling and mineralization.

Animal models show that vitamin D deficiency may have severe consequences for skeletal health. However, most studies have been performed in young rodents for a relatively short period, while in older adult rodents the effects of long-term vitamin D deficiency on skeletal health have not been extensively studied. Therefore, the first aim of this study was to determine the effects of long-term vitamin D deficiency on bone structure, remodeling and mineralization in bones from older adult mice. The second aim was to determine the effects of long-term vitamin D deficiency on mRNA levels of genes involved in vitamin D metabolism in bones from older adult mice. Ten months old male C57BL/6 mice were fed a diet containing 0.5% calcium, 0.2% phosphate and 0 (n=8) or 1 (n=9) IU vitamin D3/gram for 14 months. At an age of 24 months, mice were sacrificed for histomorphometric and micro-computed tomography (micro-CT) analysis of humeri as well as analysis of CYP27B1, CYP24 and VDR mRNA levels in tibiae and kidneys using RT-qPCR. Plasma samples, obtained at 17 and 24 months of age, were used for measurements of 25-hydroxyvitamin D (25(OH)D) (all samples), phosphate and parathyroid hormone (PTH) (terminal samples) concentrations. At the age of 17 and 24 months, mean plasma 25(OH)D concentrations were below the detection limit (<4nmol/L) in mice receiving vitamin D deficient diets. Plasma phosphate and PTH concentrations did not differ between both groups. Micro-CT and histomorphometric analysis of bone mineral density, structure and remodeling did not reveal differences between control and vitamin D deficient mice. Long-term vitamin D deficiency did also not affect CYP27B1 mRNA levels in tibiae, while CYP24 mRNA levels in tibiae were below the detection threshold in both groups. VDR mRNA levels in tibiae from vitamin D deficient mice were 0.7 fold lower than those in control mice. In conclusion, long-term vitamin D deficiency in older adult C57BL/6 mice, accompanied by normal plasma PTH and phosphate concentrations, does not affect bone structure, remodeling and mineralization. In bone, expression levels of CYP27B1 are also not affected by long-term vitamin D deficiency in older adult C57BL/6 mice. Our results suggest that mice at old age have a low or absent response to vitamin D deficiency probably due to factors such as a decreased bone formation rate or a reduced response of bone cells to 25(OH)D and 1,25(OH)2D. Older adult mice may therefore be less useful for the study of the effects of vitamin D deficiency on bone health in older people.

Cholecystokinin regulates satiation independently of the abdominal vagal nerve in a pig model of total subdiaphragmatic vagotomy.

The vagal nerve and gut hormones CCK and GLP-1 play important roles in the control of food intake. However, it is not clear to what extent CCK and GLP-1 increase satiation by stimulating receptors located on abdominal vagal nerve endings or via receptors located elsewhere. This study aimed to further explore the relative contribution of the abdominal vagal nerve in mediating the satiating effects of endogenous CCK and GLP-1. Total subdiaphragmatic vagotomy or sham operation was combined with administration of CCK1 and GLP-1 receptor antagonists devazepide and exendin (9-39) in 12 pigs, applying an unbalanced Latin Square within-subject design. Furthermore, effects of vagotomy on preprandial and postprandial acetaminophen absorption, glucose, insulin, GLP-1 and CCK plasma concentrations were investigated. Ad libitum liquid meal intake (mean±SEM) was similar in sham and vagotomized pigs (4180±435 and 3760±810 g/meal). Intake increased by about 20% after blockade of CCK1 receptors, independently of the abdominal vagal nerve. Food intake did not increase after blockade of GLP-1 receptors. Blockade of CCK1 and GLP-1 receptors increased circulating CCK and GLP-1 concentrations in sham pigs only, suggesting the existence of a vagal reflex mechanism in the regulation of plasma CCK1 and GLP-1 concentrations. Vagotomy decreased acetaminophen absorption and changed glucose, insulin, CCK and GLP-1 concentrations indicating a delay in gastric emptying. Our data show that at liquid feeding, satiation is decreased effectively by pharmacological blockade of CCK1 receptors. We conclude that regulation of liquid meal intake appears to be primarily regulated by CCK1 receptors not located on abdominal vagal nerve endings.

Microvascular changes in aged rat forebrain. Effects of chronic nimodipine treatment.

In the present study the effects of long-term treatment with the 1,4-dihydropyridine calcium antagonist nimodipine on ultrastructural alterations of the microvascular morphology were examined in the frontoparietal cortex, entorhinal cortex and CA1 of the hippocampus in the aged rat. Qualitative observations of cerebral microvasculature of aged (30 months) Wistar rats revealed the presence of microvascular fibrosis, membranous inclusions within the basement membrane and basement membrane thickenings. In several cortical regions the percentage of aberrant microvessels was significantly reduced in the nimodipine-treated rats. The observed microvascular anomalies were classified into five distinct categories of which microvascular fibrosis type II, defined as collagen deposits up to 1 micron within the microvascular basement membrane, showed the strongest reduction in the nimodipine-treated cases. The decrement of the percentage of aberrant microvessels and the relative occurrence of several classes of microvascular deviations showed some variation in the various brain regions examined and was most pronounced in frontoparietal cortex layer III. These results may provide a morphological basis for the improved motor and cognitive performance in aged rats after long-term oral nimodipine administration.

Beneficial effect of nimodipine on peripheral nerve function in aged rats.

In aged rats neuromuscular function and motor coordination is gradually impaired. Major motor deficits were seen in rats of more than 2 years of age; with increasing age, the incidence of abnormal footprints increased sharply. Oral nimodipine, a Ca2(+)-entry blocker of the dihydropyridine type, treatment suppressed and/or delayed the appearance of these abnormal footprints. In aged rats that already displayed a considerable amount of abnormal footprints in the free walking pattern, oral nimodipine treatment was similarly effective. Nimodipine not only delays the onset of age-related motor deficits, but also may counteract these deficits once already present. In aged rats the nerve conduction velocities were severely diminished. Nimodipine treatment resulted in an enhancement of the sciatic and caudal nerve conduction velocities. Histological analysis revealed a lower fiber density in aged rats compared to aged nimodipine-treated rats. Whether nimodipine acts directly on the peripheral nervous system is currently unclear. Nevertheless, the present study lends further support for the beneficial effects of nimodipine in age-related motor deficits in the rat.

Protein kinase C activity, translocation, and conventional isoforms in aging rat brain.

Protein kinase C was studied in various brain areas in aging Wistar rats. Histone-directed kinase activity from the cortex, hippocampus and cerebellum did not change with aging. Using purified protein B-50 as a substrate, between 3 and 8 months a decrease in in vitro phosphorylation was detected in the membrane fraction of the cortex but after this age values remained stable. In hippocampal membranes, B-50 phosphorylation was increased in aged rats. PKC translocation was impaired in aged rats in both the cortex and the hippocampus. PKC alpha and beta mRNA decreased in the cortex between 3 and 8 months with no further decline in aged animals. Hippocampal mRNA for calcium-dependent PKC isoforms was not modified during aging, as assessed by Northern and in situ hybridization. Western blot analysis revealed a change in PKC gamma protein only, which was increased in hippocampal membranes from aged rats. The data indicate that the key PKC function that is impaired in aged rats is enzyme translocation irrespective of the brain area investigated.

Impaired Learning and Abnormal Open-field Behaviours of Rats After Early Postnatal Anoxia and the Beneficial Effect of the Calcium Antagonist Nimodipine.

Perinatal anoxia/hypoxia is considered a serious risk factor for normal brain development. Anoxia induced by repeated asphyxia at 2 and 4 days after birth resulted in a transient hyperactivity in the small open-field, and a behavioural depression in adult open-field activity of male Wistar rats. The same treatment impaired adult learning behaviour in pole-jumping conditioned avoidance and appetitively motivated hole-board test situations. The calcium entry blocker nimodipine (in doses of 3 and 10 mg/kg) prevented the anoxia-induced changes in orientation motility in the open-field tests and almost fully antagonized the learning deficit in the hole-board test. The behavioural deficit seen during acquisition of the pole-jumping conditioned avoidance response was ameliorated to a lesser degree. The results indicate that the maintenance of calcium homeostasis during the early postnatal phase of brain development is crucial to prevent anoxia-induced behavioural abnormalities.

Neuropeptide substance P improves water maze performance in aged rats.

The effects of the peripherally administered neuropeptide substance P (SP) on spatial learning capacities were investigated in 27-month-old rats using a water-maze task. Old rats were injected intraperitoneally once daily for 6 days with 50 or 250 micrograms/kg SP or vehicle 30 min prior to acquisition trials. Improvement in maze performance was observed following injections of 250 micrograms/kg SP only. Furthermore, vehicle-treated old rats showed significantly poorer acquisition rates than vehicle-treated 12-week-old rats. Thus, the improvement in performance after the 250 micrograms/kg dose of SP can be interpreted in terms of a compensation of performance deficit in the old rats.

Cerebrovascular, neuronal, and behavioral effects of long-term Ca2+ channel blockade in aging normotensive and hypertensive rat strains.

The pathogenesis of essential hypertension is not fully understood, but most of the cardiovascular, metabolic, neurogenic, and humoral abnormalities are explained by dysfunctions in the control of intracellular Ca2+ concentrations in the cells of the vascular wall. Most theories of disturbed calcium regulation focus on the calcium concentration within vascular smooth muscle cells. The implications of hypertension for the increased calcium content of aging arteries seem to be clear, but were only studied in the peripheral circulation; hypertension prominently augments the aging-related accumulation of calcium in the vessel wall. Although the contribution of calcium overload in hypertensive cerebrovascular damage is well documented, it is not clear yet if hypertension per se is the main cause of hypertension-associated calcium-dependent cerebral damage. Thus far, the hypotensive effects of most calcium antagonists were extensively described, and their efficacy in stroke prevention was proven. Earlier studies indicated that chronic administration of nimodipine revealed a protective effect in the occurrence of strokes in SHR-SP rats, yielding a decreased mortality rate. Because nimodipine did not lower the extremely high blood pressure of these animals, the mechanisms behind such nimodipine-induced stroke prevention may be attributed to a direct cerebrovascular and/or neuronal action of nimodipine. Hypertension is generally considered a vascular pathologic condition, and most research has been directed towards the influences of hypertension on large peripheral arteries such as the aorta and coronary artery. The influence of the CNS on the regulation of cardiovascular system and blood pressure regulation was described in detail, and the role of the CNS in hypertension also was the subject of study. The increased risk of stroke in hypertensive subjects generated numerous studies on the precise nature of compromised cerebrovascular functioning under hypertensive conditions. Few data are available on Ca2+ alterations in cerebral neurons during hypertension. Honda et al. demonstrated that voltage-dependent Ca2+ uptake was higher in cortical synaptosomes from SHR than form normotensive animals and suggested that an important alteration in Ca2+ channel characteristics may occur in SHR brain synaptosomes. Although the density of L-type calcium channels was shown to be higher in the hippocampus of SHR rats, others reported that the number of L-type calcium channels was significantly lower in the brain of SHR rats than WKY normotensive controls. The latter data suggest that hypertension may be associated with similar alterations in neuronal calcium homeostasis as demonstrated for aging in normotensive subjects.(ABSTRACT TRUNCATED AT 400 WORDS)

Protective effect of the calcium antagonist nimodipine on discrimination learning deficits and impaired retention behavior caused by prenatal nitrite exposure in rats.

Discrimination learning behavior and retention of a passive avoidance response were studied in male adult offspring of gestating rats exposed to drinking water containing 2 g/l sodium nitrite, throughout the second half of pregnancy. Both in an auditory and visual discrimination learning paradigm NaNO2-exposed rats were inferior to controls. The long-term retention of a passive avoidance response was also impaired. The acquisition of simple learning tasks was not significantly disturbed. The concomitant prenatal daily treatment with the calcium antagonist nimodipine in a dose of 10 mg/kg p.o. interfered with the nitrite neurotoxicity and prevented the development of adult behavioral deficits. The results support the hypothesis that Ca2+ homeostasis of neurons is an important factor for normal development of brain and behavior.

Synaptic plasticity in the dentate gyrus of aged rats is altered after chronic nimodipine application.

We examined ultrastructural correlates of synaptic plasticity in the hippocampus of young (3 months) vs aged (30 months) Wistar rats and established the effects of the calcium antagonist nimodipine in animals chronically treated from 24 to 30 months. The effects of nimodipine was studied since this compound improves hippocampal neuronal physiology and enhances cognitive function during aging. In the supragranular layer of the dentate gyrus we found a 24% decrease in synaptic density (Nv) in aged animals, while synaptic size (S) was not significantly altered. After nimodipine treatment Nv in aged rats was not significantly different from young adults, thus being significantly increased compared to age-matched controls. The size of synapses was not significantly altered after nimodipine administration. Total synaptic surface area (Sv) in nimodipine-treated animals was significantly increased compared to aged controls, however, Sv remained significantly lower than in young adults. These data indicate that chronic administration of nimodipine enables granular cells in the dentate gyrus to maintain its number of synaptic contacts during the aging process. Furthermore, the presented influence of nimodipine on synaptic plasticity processes may underlie previously reported improved cognitive functioning of aged animals treated similarly with nimodipine.

Nembutal treatment of the VMH (rat): effects on feeding and sexual behaviour.

In male rats two brain cannulae were implanted bilaterally and directed to an area just dorsal of the ventromedial hypothalamic nucleus. The behaviour of these animals was observed before and after the injection through these cannulae of either 1 micronl saline or 1 micronl Nembutal. Injections were performed under three behavioural conditions: (1) animal alone in cage with food, (2) animal in the presence of an oestrous female and (3) animal in cage with food and oestrous female. Following the Nembutal injection, sniffing and feeding are disinhibited temporarily in condition 1, whereas in condition 3 only sniffing, but not feeding, is disinhibited. In conditions 2 and 3 male sexual behaviour is not influenced by the Nembutal except for an increase of sniffing at the female. It is concluded that a temporary elimination of the VMH leads to a disinhibition of feeding only if the external conditions are favourable for feeding. The same elimination of the VMH does not influence the occurrence of male sexual behaviour under the conditions used in these experiments.

Anxiolytics and stress-induced behavioural and cardiac responses: a study of diazepam and ipsapirone (TVX Q 7821).

The present study has been designed to investigate the effects of the 5-HT1A receptor agonist, ipsapirone (TVX Q 7821), a representative of a novel class of anxiolytics, and the classical benzodiazepine anxiolytic, diazepam, on cardiac and behavioural responses in an emotional stress situation. The emotional stress of fear of punishment, induced by training male Wistar rats in an inhibitory avoidance situation, was followed by a bradycardiac response relative to similarly trained, but non-punished, freely moving rats. The behavioural response of stressed rats was immobility in the dark compartment in which an electric footshock (0.6 mA a.c. for 3 s) had been administered as punishment a day earlier. Diazepam administered i.p. in doses of 2.5 mg/kg and 7.5 mg/kg caused a decrease in the interbeat interval (IBI) in shocked and non-shocked rats whereas ipsapirone administered i.p. in doses of 2.5 and 12.5 mg/kg decreased the IBI in shocked rats only. Ipsapirone diminished the duration of immobility in both shocked and non-shocked animals whereas diazepam decreased immobility in shocked rats only. These results suggest a differential effect of the two anxiolytics on the behavioural and cardiac responses to an emotional stress situation. It is suggested that ipsapirone has an anxiolytic-like effect and 'anti-stress' action that is clearly reflected in the cardiac physiology in an anxiety-inducing situation.