2-arachidonoylglycerol signaling impairs short-term fear extinction.

Impairments in fear extinction are thought to be central to the psychopathology of posttraumatic stress disorder, and endocannabinoid (eCB) signaling has been strongly implicated in extinction learning. Here we utilized the monoacylglycerol lipase inhibitor JZL184 to selectively augment brain 2-AG levels combined with an auditory cue fear-conditioning paradigm to test the hypothesis that 2-AG-mediated eCB signaling modulates short-term fear extinction learning in mice. We show that systemic JZL184 impairs short-term extinction learning in a CB1 receptor-dependent manner without affecting non-specific freezing behavior or the acquisition of conditioned fear. This effect was also observed in over-conditioned mice environmentally manipulated to re-acquire fear extinction. Cumulatively, the effects of JZL184 appear to be partly due to augmentation of 2-AG signaling in the basolateral nucleus of the amygdala (BLA), as direct microinfusion of JZL184 into the BLA produced similar results. Moreover, we elucidate a short ~3-day temporal window during which 2-AG augmentation impairs extinction behavior, suggesting a preferential role for 2-AG-mediated eCB signaling in the modulation of short-term behavioral sequelae to acute traumatic stress exposure.

Canine breed predispositions for marked hypocobalaminaemia or decreased folate concentration assessed by a laboratory survey.

OBJECTIVE: The aim of this study was to determine canine breed predispositions for decreased serum folate or markedly decreased cobalamin concentrations. METHODS: Retrospective analysis of samples from dogs that had serum folate and cobalamin concentrations measured, from 1990 to 2002 at the Comparative Gastroenterology Laboratory of Liverpool, were enrolled. A total of 13,069 samples were analysed. Those with trypsin-like immunoreactivity < 5·0 lg/L were excluded, and only breeds with at least 30 individuals tested were further analyzed. Breed predisposition was determined by calculating odds ratios and 95% confidence intervals for hypocobalaminaemia or decreased folate concentration. Significance was tested with a two-sided Fisher's exact test, and the level of statistical significance was set at P<0·05. RESULTS: A total of 9960 dogs fulfilled the inclusion criteria. Forty breeds contained at least 30 individuals. Predispositions for hypocobalaminaemia were identified in shar peis, Staffordshire bull terriers, German shepherd dogs and mixed breeds. Predispositions for decreased folate concentration were found in golden retrievers and boxers. CLINICAL SIGNIFICANCE: Predisposition for marked hypocobalaminaemia and decreased folate concentration differed between breeds. The shar peis had a markedly increased odds ratio for hypocobalaminaemia, as previously reported in North America, but other at-risk breeds were also identified.

The role of mu-opioid receptor signaling in the dorsolateral periaqueductal gray on conditional and unconditional responding to threatening and aversive stimuli.

Here we examined how mu-opioid receptor signaling in the periaqueductal gray (PAG) mediates conditional and unconditional responses to aversive stimuli. The mu-opioid agonist morphine (MOR) and/or the partially mu-selective antagonist naltrexone (NAL) were infused into dorsolateral PAG (dlPAG) during a fear conditioning task, in which rats were trained to fear an auditory conditional stimulus (CS) by pairing it with a unilateral eyelid shock unconditional stimulus (US). During drug-free test sessions, the CS elicited movement suppression responses (indicative of freezing) from trained rats that had not recently encountered the US. In trained rats that had recently encountered the US, the CS elicited flight behavior characterized by turning in the direction away from the eyelid where US delivery was anticipated. Infusions of MOR (30 nmol/side) into dlPAG prior to the test session did not impair CS-evoked movement suppression, but did impair CS-evoked turning behaviors. MOR infusions also reduced baseline motor movement, but US-evoked reflex movements remained largely intact. NAL was infused at two dosages, denoted 1x (26 nmol/side) and 10x (260 nmol/side). Infusions of NAL into dlPAG did not affect CS- or US-evoked behavioral responses at the 1x dosage, but impaired CS-evoked movement suppression at the 10x dosage, both in the presence and absence of MOR. When rats were co-infused with MOR and NAL, MOR-induced effects were not reversed by either dosage of NAL, and some measures of MOR-induced movement suppression were enhanced by NAL at the 1x dosage. Based on these findings, we conclude that mu-opioid receptors in dlPAG may selectively regulate descending supraspinal motor pathways that drive active movement behaviors, and that interactions between MOR and NAL in dlPAG may be more complex than simple competition for binding at the mu receptor.

The effect of silver impregnation of surgical scrub suits on surface bacterial contamination.

Silver-impregnated fabrics are widely used for their antibacterial and antifungal effects, including for clinical clothing such as surgical scrub suits (scrubs). This study investigated whether silver impregnation reduces surface bacterial contamination of surgical scrubs during use in a veterinary hospital. Using agar contact plates, abdominal and lumbar areas of silver-impregnated nylon or polyester/cotton scrubs were sampled for surface bacterial contamination before (0 h) and after 4 and 8h of use. The number of bacterial colonies on each contact plate was counted after 24 and 48 h incubation at 37°C. Standard basic descriptive statistics and mixed-effects linear regression were used to investigate the association of possible predictors of the level of bacterial contamination of the scrubs with surface bacterial counts. Silver-impregnated scrubs had significantly lowered bacterial colony counts (BCC) at 0 h compared with polyester/cotton scrubs. However, after 4 and 8h of wear, silver impregnation had no effect on BCC. Scrub tops with higher BCC at 0 h had significantly higher BCC at 4 and 8h, suggesting that contamination present at 0 h persisted during wear. Sampling from the lumbar area was associated with lower BCC at all three time points. Other factors (contamination of the scrub top with a medication/drug, restraint of patients, working in the anaesthesia recovery area) also affected BCC at some time points. Silver impregnation appeared to be ineffective in reducing bacterial contamination of scrubs during use in a veterinary hospital.

Conditioned turning behavior: a Pavlovian fear response expressed during the post-encounter period following aversive stimulation.

Rats were trained to fear an auditory conditioned stimulus (CS) by pairing it with a mild electric shock (the unconditioned stimulus, or US) delivered to one eyelid. After training, the CS elicited two different conditioned fear responses from rats: a passive freezing response, and an active turning response. The balance between these two modes of conditioned responding depended upon the rat's recent history of encounters with the US. If rats had not recently encountered the US, then they responded to the CS by freezing. But after recently encountering the US, rats exhibited CS-evoked turning responses that were always directed away from the trained eyelid, even if the US had recently been delivered to the opposite (untrained) eyelid. This post-encounter turning behavior was not observed in rats that had been trained with unpaired presentations of the CS and US, indicating that even though CS-evoked turning was selectively expressed after recent encounters with the US, it was nonetheless a conditioned Pavlovian fear response that depended upon a learned association between the CS and US. Further supporting this conclusion, pharmacological inactivation experiments showed that expression of both freezing and turning behaviors depended upon lateralized circuits in the amygdala and periaqueductal gray (PAG) that are known to support expression of Pavlovian fear responses. These findings indicate that even though the ability of a CS to elicit Pavlovian fear responses depend upon the long-term history of CS-US pairings, the mode of conditioned responding (freezing versus turning in the present experiments) can be modulated by short-term factors, such as the recent history of US encounters. We discuss neural mechanisms that might mediate such short-term transitions between different modes of defensive responding, and consider how dysregulation of such mechanisms might contribute to clinical anxiety disorders.

First-choice therapy for dogs presenting with diarrhoea in clinical practice.

Computerised referral histories were reviewed for dogs admitted to the University of Liverpool Small Animal Teaching Hospital between January 2000 and December 2008 with diarrhoea among the clinical signs. A total of 371 cases presenting to the referring veterinary surgeon were included in the study, and information was compiled regarding signalment, clinical signs and treatment given at the initial consultation. Various breeds, ages and sexes were represented. Antibacterials were used in 263 (71 per cent) cases, steroids in 71 (19 per cent) cases and miscellaneous antidiarrhoeal products (including probiotics, prebiotics, adsorbents and antimotility drugs) in 98 (26 per cent) cases. Other drugs used included antiemetics (48 of 371 [13 per cent] cases), gastric protectants (37 of 371 [10 per cent] cases) and sulfasalazine (26 of 371 [7 per cent] cases). Antibacterial administration was positively associated with hyperthermia (odds ratio [OR]=2.97, P=0.012) and anorexia (OR=2.17, P=0.0075), but negatively associated with both weight loss (OR=0.55, P=0.036) and tenesmus (OR=0.43, P=0.035). In contrast, use of antidiarrhoeal products was positively associated with the presence of faecal mucus (OR=1.77, P=0.043), and negatively associated with vomiting (OR=0.57, P=0.025) and weight loss (OR=0.52, P=0.033).

Bilateral phosphorylation of ERK in the lateral and centrolateral amygdala during unilateral storage of fear memories.

We previously showed that when rats were trained to fear an auditory conditioned stimulus (CS) by pairing it with a mild unilateral shock to the eyelid (the unconditioned stimulus, or US), conditioned freezing depended upon the amygdala contralateral but not ipsilateral from the US. It was proposed that convergent activation of amygdala neurons by the CS and US occurred mainly in the amygdala contralateral from US delivery, causing memories of the CS-US association to be stored primarily by that hemisphere. In the present study, we further tested this interpretation by administering unilateral infusions of U0126 (in 50% dimethyl sulfoxide (DMSO) vehicle) to block phosphorylation of extracellular signal-responsive kinase (ERK) in the amygdala prior to CS-US pairings. Conditioned freezing was impaired 24 h after training when U0126 was infused contralaterally-but not ipsilaterally-from the US, suggesting that fear memories were consolidated mainly by the contralateral amygdala. However, immunostaining experiments revealed that ERK phosphorylation was elevated in both hemispheres of the amygdale's lateral (LA) and centrolateral (CeL) nuclei after paired (but not unpaired (UNP)) presentations of the CS and US. Thus, fear acquisition induced ERK phosphorylation bilaterally in the amygdala, even though the ipsilateral hemisphere did not appear to participate in conditioned freezing. These findings suggest that associative plasticity may occur in both amygdala hemispheres even when only one hemisphere is involved in freezing behavior. Conditioning-induced ERK phosphorylation was identical in both hemispheres of LA, but was slightly greater in the contralateral than ipsilateral hemisphere of CeL. Hence, asymmetric induction of plasticity in CeL might help to explain why conditioned freezing depends preferentially upon the amygdala contralateral from the US in our fear conditioning paradigm.

Effects of dorsal striatal infusions of R(-)-propylnorapomorphine on kappa-opioid-mediated locomotor activity in the young rat: possible role of the indirect pathway.

Stimulation of kappa-opioid receptors in the substantia nigra pars reticulata (SNPR) increases the locomotor activity of young rats: an effect blocked by systemic administration of a D2-like receptor agonist. Based on these initial findings, we proposed that: (a) D2-like receptors in the dorsal striatum are responsible for attenuating kappa-opioid-induced locomotor activity, and (b) the effects of D2-like receptor stimulation are mediated by the indirect pathway, which extends from the dorsal striatum to the SNPR via the globus pallidus (GP) and subthalamic nucleus (STN). To test the first hypothesis, young rats were given a systemic injection (i.p.) of saline or the kappa-opioid receptor agonist (+/-)-trans-U50,488 methanesulfonate salt (U50,488) on postnatal day (PD) 18. Later in the testing session, rats received bilateral infusions of vehicle or the D2-like receptor agonist R(-)-propylnorapomorphine (NPA) into the dorsal striatum, and the ability of NPA to block U50,488-induced locomotor activity was determined. To test the second hypothesis, rats were given sham or bilateral electrolytic lesions of the GP or STN on PD 16. Two days later, saline- and U50,488-induced locomotor activity was measured after systemic (i.p.) administration of vehicle or NPA. As predicted, dorsal striatal infusions of NPA attenuated the U50,488-induced locomotor activity of young rats. Contrary to our expectations, bilateral lesions of the GP or STN did not impair NPA's ability to block U50,488-induced locomotor activity. When considered together, these results suggest that: (a) stimulation of D2-like receptors in the dorsal striatum is sufficient to attenuate the kappa-opioid-mediated locomotor activity of young rats; and (b) the indirect pathway does not mediate the effects of D2-like receptor stimulation in this behavioral model.