11 Beta-hydroxysteroid dehydrogenase type 1 regulates synovitis, joint destruction, and systemic bone loss in chronic polyarthritis.

OBJECTIVE: In rheumatoid arthritis, the enzyme 11 beta-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) is highly expressed at sites of inflammation, where it converts inactive glucocorticoids (GC) to their active counterparts. In conditions of GC excess it has been shown to be a critical regulator of muscle wasting and bone loss. Here we examine the contribution of 11ß-HSD1 to the pathology of persistent chronic inflammatory disease. METHODS: To determine the contribution of 11ß-HSD1 to joint inflammation, destruction and systemic bone loss associated with persistent inflammatory arthritis, we generated mice with global and mesenchymal specific 11ß-HSD1 deletions in the TNF-transgenic (TNF-tg) model of chronic polyarthritis. Disease severity was determined by clinical scoring. Histology was assessed in formalin fixed sections and fluorescence-activated cell sorting (FACS) analysis of synovial tissue was performed. Local and systemic bone loss were measured by micro computed tomography (micro-CT). Measures of inflammation and bone metabolism were assessed in serum and in tibia mRNA. RESULTS: Global deletion of 11ß-HSD1 drove an enhanced inflammatory phenotype, characterised by florid synovitis, joint destruction and systemic bone loss. This was associated with increased pannus invasion into subchondral bone, a marked polarisation towards pro-inflammatory M1 macrophages at sites of inflammation and increased osteoclast numbers. Targeted mesenchymal deletion of 11ß-HSD1 failed to recapitulate this phenotype suggesting that 11ß-HSD1 within leukocytes mediate its protective actions in vivo. CONCLUSIONS: We demonstrate a fundamental role for 11ß-HSD1 in the suppression of synovitis, joint destruction, and systemic bone loss. Whilst a role for 11ß-HSD1 inhibitors has been proposed for metabolic complications in inflammatory diseases, our study suggests that this approach would greatly exacerbate disease severity.

Cranial ultrasound for the diagnosis of giant cell arteritis. A retrospective cohort study.

BACKGROUND: Establishing a diagnosis of giant cell arteritis, or indeed ruling it out, may be difficult. We describe an evaluation of temporal artery colour duplex ultrasound as first line investigation in patients with suspected giant cell arteritis. METHODS: A retrospective cohort study of all patients undergoing colour duplex ultrasound for suspected giant cell arteritis between January 2005 and January 2014 was undertaken at a teaching hospital. A minimum clinical follow-up of three months was required. Patients were classified on the basis of ultrasound reports, using described features such as a halo sign or arterial wall thickening and clinical diagnosis of giant cell arteritis after at least 3 months follow-up, determined by the treating physician. The relationship of colour duplex ultrasound to a final clinical diagnosis of giant cell arteritis was analysed. RESULTS: A total of 87 patients underwent colour duplex ultrasound: 36 (41%) had clinically confirmed giant cell arteritis at 3-month follow-up. The positive predictive value of colour duplex ultrasound for a clinical diagnosis at 3 months was 97% (95% confidence interval (CI) 93 to 99%) and negative predictive value 88% (95% CI 76 to 95%). Sensitivity was 81% (95% CI 64 to 92%) and specificity 98% (95% CI 90 to 100%). CONCLUSIONS: A high positive and negative predictive value of arteritis on colour duplex ultrasound indicates that temporal artery biopsy may be unnecessary in suspected giant cell arteritis, particularly where clinical suspicion of giant cell arteritis is high or low.

Paradoxical coronary artery embolism causing acute myocardial infarction in a young woman with factor V Leiden thrombophillia.

Paradoxical coronary artery embolism is a rare but under-diagnosed cause of acute myocardial infarction (AMI) and requires a high level of clinical suspicion to make an early diagnosis. We describe the case of a young woman who presented with a severe cough and chest pain who was subsequently found to have a paradoxical embolus in the right coronary artery. Echocardiography showed a patent foramen ovale (PFO) and an atrial septal aneurysm (ASA). The patient was found to be a heterozygous carrier of the factor V Leiden mutation that increases the risk for venous-thromboembolism. The association between a PFO and an ASA is a risk factor for systemic embolisation. This is the first reported case of paradoxical coronary artery embolus causing AMI in a non-pregnant patient with factor Leiden thrombophilia. Identification of this clinical phenotype is vital as the risk of future embolic events can be reduced by anticoagulation and closure of anatomical cardiac defects.

Nimodipine prior to alcohol withdrawal prevents memory deficits during the abstinence phase.

Effects of the dihydropyridine, nimodipine, an antagonist at L-type calcium channels, on the memory loss in rats caused by long term alcohol consumption were examined. Either a single dose of nimodipine or 2 weeks of repeated administration was given prior to withdrawal from 8 months of alcohol consumption. Memory was measured by the object recognition test and the T maze. Both nimodipine treatments prevented the memory deficits when these were measured between 1 and 2 months after alcohol withdrawal. At the end of the memory testing, 2 months after cessation of chronic alcohol consumption, glucocorticoid concentrations were increased in specific regions of rat brain without changes in plasma concentrations. Both nimodipine treatment schedules substantially reduced these rises in brain glucocorticoid. The data indicate that blockade of L-type calcium channels prior to alcohol withdrawal protects against the memory deficits caused by prolonged alcohol intake. This shows that specific drug treatments, such as nimodipine, given over the acute withdrawal phase, can prevented the neuronal changes responsible for subsequent adverse effects of long term consumption of alcohol. The results also suggest the possibility that regional brain glucocorticoid increases may be involved in the adverse effects of long term alcohol intake on memory. Such local changes in brain glucocorticoid levels would have major effects on neuronal function. The studies indicate that L-type calcium channels and brain glucocorticoid levels could form new targets for the treatment of cognitive deficits in alcoholics.

Selective increases in regional brain glucocorticoid: a novel effect of chronic alcohol.

The hypothalamo-pituitary-adrenal axis shows functional changes in alcoholics, with raised glucocorticoid release during alcohol intake and during the initial phase of alcohol withdrawal. Raised glucocorticoid concentrations are known to cause neuronal damage after withdrawal from chronic alcohol consumption and in other conditions. The hypothesis for these studies was that chronic alcohol treatment would have differential effects on corticosterone concentrations in plasma and in brain regions. Effects of chronic alcohol and withdrawal on regional brain corticosterone concentrations were examined using a range of standard chronic alcohol treatments in two strains of mice and in rats. Corticosterone was measured by radioimmunoassay and the identity of the corticosterone extracted from brain was verified by high performance liquid chromatography and mass spectrometry. Withdrawal from long term (3 weeks to 8 months) alcohol consumption induced prolonged increases in glucocorticoid concentrations in specific regions of rodent brain, while plasma concentrations remained unchanged. This effect was seen after alcohol administration via drinking fluid or by liquid diet, in both mice and rats and in both genders. Shorter alcohol treatments did not show the selective effect on brain glucocorticoid levels. During the alcohol consumption the regional brain corticosterone concentrations paralleled the plasma concentrations. Type II glucocorticoid receptor availability in prefrontal cortex was decreased after withdrawal from chronic alcohol consumption and nuclear localization of glucocorticoid receptors was increased, a pattern that would be predicted from enhanced glucocorticoid type II receptor activation. This novel observation of prolonged selective increases in brain glucocorticoid activity could explain important consequences of long term alcohol consumption, including memory loss, dependence and lack of hypothalamo-pituitary responsiveness. Local changes in brain glucocorticoid levels may also need to be considered in the genesis of other mental disorders and could form a potential new therapeutic target.

Social defeat increases alcohol preference of C57BL/10 strain mice; effect prevented by a CCKB antagonist.

RATIONALE: In humans, social stress over long and short term can increase alcohol consumption, but the mechanisms involved are not understood. OBJECTIVES: This study was conducted to examine the effects of social defeat, using the resident/intruder paradigm, on the alcohol preference of "low alcohol drinking" individuals in a colony of C57BL/10 strain mice and the effects of two anxiolytic drugs. METHODS: Alcohol preference, in a two-bottle choice (8% v/v alcohol or water), was measured, in separate experiments, after either a single experience of social defeat by a resident male mouse, five consecutive daily defeat experiences or one experience per week for 4 weeks. Comparison was made with effects of repeated social defeat on the preference for dilute sucrose. In addition, the actions of the CCKB receptor antagonist, CAM1028, and of diazepam were examined on the effects of repeated defeat experiences. RESULTS: Five consecutive daily defeat experiences had a slow onset effect in increasing alcohol preference and consumption, compared with five daily exposures to a novel environment. A single defeat, or one defeat per week, did not significantly alter alcohol preference or intake. There were no effects of five daily defeat experiences on sucrose preference or consumption. The effect of repeated defeats on alcohol preference was significantly decreased by administration of the CCKB receptor antagonist, CAM1028, prior to each experience, but not by corresponding administration of diazepam. CONCLUSION: The results show that social stress increases alcohol intake in low alcohol preference C57BL/10 mice and suggest that CCK transmission may be involved in this effect.

Generation of neuroprogenitor-like cells from adult mammalian bone marrow stromal cells in vitro.

Recently, it has been proposed that bone marrow stromal cells (BMSCs) have a broader capacity for differentiation than previously contemplated. In vitro studies have indicated that BMSCs may have the capacity to differentiate into neuroectodermal-like cells in response to various growth conditions, including those commonly used to maintain and differentiate cultures of primary neural stem cells (NSCs). Interpreting the wealth of data on this subject has been difficult because of variation in the starting cell population and the differences between the methods used to induce their differentiation. Here we evaluate how cultures of expanded BMSCs with a consistent immunophenotype respond to a variety of growth conditions and induction agents and review their ability to form neural-like derivatives. In addition, we report on some modifications to previously published techniques for the generation of neural-like cells from BMSCs in vitro.

Effects of corticosterone on place conditioning to ethanol.

RATIONALE AND OBJECTIVE: Effects of corticosterone on place conditioning to ethanol were investigated in mice using two conditioning schedules; the conventional method and a rapid conditioning schedule in which exposure to the CS+ followed immediately on exposure to the CS-. METHODS: Effects of administration of corticosterone, 10 mg/kg, on the acquisition of place conditioning produced by ethanol, 1-2.5 g/kg, were investigated using the conventional method of conditioning, with exposure to the CS+ and the CS- on alternate days, and also using the rapid conditioning method. Total and free blood corticosterone concentrations were measured after administration of ethanol and corticosterone. RESULTS: In the conventional, alternate day, conditioning schedule, ethanol produced significant place preference at 2 and at 2.5 g/kg, but when these alcohol doses were given with corticosterone 10 mg/kg, significant place conditioning was not seen. In contrast, in the rapid, same day, conditioning schedule corticosterone significantly decreased the dose at which ethanol produced an apparent place preference, with significant place conditioning being seen with ethanol at 1 and 1.5 g/kg in combination with corticosterone, 10 mg/kg. Total and free corticosterone concentrations were increased after ethanol, 1.5 g/kg, compared with controls, and administration of corticosterone, 10 mg/kg, caused a significantly greater increase. There were no significant differences in spontaneous locomotor activity or brain alcohol concentrations between any of the treatment groups. CONCLUSIONS: The effects of corticosterone on ethanol-induced place conditioning are substantially affected by the conditioning schedule used.

Low alcohol preference among the "high alcohol preference" C57/BL10 mice; factors affecting such preference.

The effects of age, ethanol concentration and minor stress on the variation in alcohol preference of C57 strain mice were determined. In two bottle choice tests, an older population of mice contained slightly more low-preference mice than a younger population. A wide range of ethanol preference was consistently seen in young mice for 8% and 6% ethanol, but the previously reported biphasic pattern of distribution was revealed only with 8% ethanol. Very few animals showed high preference for concentrations of 10% or 12% ethanol. Moving low alcohol preference mice to a new location (but not repeated cage changing or ultrasonic noise) significantly increased the alcohol preference. Exploratory locomotor activity did not correlate with the subsequent alcohol consumption. Blood and brain alcohol concentrations showed that the differences in alcohol preference were not due to differences in metabolism of ethanol. The C57 strain mice with low preference for alcohol provides a valuable model for the study of the effects of minor stress on alcohol consumption.

Effects of intraperitoneal injections of saline on the alcohol and sucrose consumption of C57/BL10 mice.

RATIONALE: To determine the effects of multiple saline injections on alcohol drinking by male and female C57/BL10 mice with low preference for alcohol. OBJECTIVE: An investigation of the effects of multiple saline injections on alcohol consumption, with a comparison of corresponding effects on sucrose consumption. METHODS: The effects of a range of injection schedules on preference for 8% alcohol, or 1% sucrose, compared with tap water, were measured in two-bottle choice tests. RESULTS: The multiple saline injection schedule significantly increased the alcohol preference, even when no alcohol was available during the injection period. The actual administration of fluid was not necessary for the increase in alcohol preference, since sham injections without fluid administration also increased alcohol preference. A single injection of saline did not alter the alcohol preference 3 weeks later. Daily saline injections for 3 weeks did not alter the consumption of the dilute sucrose solution. In the population of mice used, the preference for sucrose over water was found to follow a biphasic distribution, similar to that reported earlier in these mice for alcohol preference, but there was no correlation between alcohol preference and sucrose preference. CONCLUSIONS: The results suggest that lasting changes in the areas of the brain that specifically control alcohol intake are produced by repetition of a routine laboratory procedure.

Alterations in mesolimbic dopamine function during the abstinence period following chronic ethanol consumption.

Previous work demonstrated that the locomotor stimulant actions of amphetamine, cocaine and nicotine were increased when these drugs were given during the abstinence phase after chronic ethanol consumption. These changes were seen at 6 days and at 2 months after cessation of alcohol. The present study examined neuronal alterations which might be related to these changes in behaviour. Markedly reduced spontaneous firing rates of dopaminergic cells in the ventral tegmental area (VTA) in midbrain slices were seen 6 days into the abstinence period after cessation of chronic ethanol consumption, but by 2 months the firing rates had returned to control values. Increased affinity of striatal receptors for the D1-like receptor ligand 3H-SCH23390, but no change in the receptor density, was found both at the 6 day and the 2 month intervals. The binding properties of striatal D2-like receptors, of D1-like and D2-like receptors in the frontal cerebral cortex, and the release of tritiated dopamine from slices of striatum or frontal cerebral cortex, were unchanged at 6 days and 2 months. It is suggested that the decreased neuronal firing leads to a persistent increase in sensitivity of D1-like receptors and that these changes could explain the increased effects of the other drugs of abuse.