The effect of infrared beak treatment on the welfare of turkeys reared to 12 weeks of age.

This study aimed to determine the effects of infrared beak treatment on the behavior and welfare of male and female turkeys reared to 12 wk of age. To do this, poults (236 males and 324 females) were assigned to one of 2 beak treatments: infrared beak treated on day of hatch (IR) or sham untreated control (C). Data collected included heterophil/lymphocyte (H/L) ratio, pecking force, feather cover, behavioral expression, and beak histology. Data were analyzed as a 2 × 2 factorial of beak treatment and gender, in a completely randomized design and analyzed using PROC MIXED (SAS 9.4). H/L ratio (indicative of a stress response) did not differ between treated and control poults during early life, except at 20 d of age when H/L ratio was higher for C poults than IR poults. Pecking force, measured as a method of monitoring pain, was different only at 1 wk of age, when IR poults pecked with more force than C poults. Feather cover was better in IR poults at 12 wk of age. Differences in behavior between treatments were minor over the 12-wk period. Overall, infrared beak treatment of commercial turkeys had minimal negative impacts on behavior and welfare. The results suggest that stress may be reduced in flocks that are beak treated and that the procedure itself does not cause a pain response.

Evaluation of the effect of vincristine on platelet count in dogs with lymphoma.

OBJECTIVES: To determine the effect of vincristine administration on the platelet count, platelet morphology and incidence of thrombocytopenia in dogs diagnosed with lymphoma. MATERIAL AND METHODS: Medical records of 59 dogs with lymphoma receiving vincristine sulphate were reviewed retrospectively. RESULTS: Following vincristine administration the platelet count was higher and the number of thrombocytopenic patients was lower. No difference was found in the number of dogs with enlarged and elliptical platelets following vincristine treatment. CLINICAL SIGNIFICANCE: Vincristine administration increases platelet counts in dogs with lymphoma. It is not contra-indicated to administer vincristine to dogs with lymphoma that are thrombocytopenic.

What has been the impact of the Traditional Herbal Registration (THR) scheme in the UK on information provided with herbal products bought over the counter?

BACKGROUND: In 2011 there was a strengthening of European Union (EU) legislation on the licencing of herbal products which, in the UK, resulted in the introduction of the Traditional Herbal Registration (THR) scheme. This scheme sets out standards for the safety and quality of herbal medicines and includes the provision of information to the customer on the safe use of the product. The aim of this study is to replicate a survey undertaken in 2011, prior to the implementation of the THR scheme, and evaluate the impact of this scheme on the information provided with herbal products bought over-the-counter. METHODS: We undertook a survey on 5 herbal products commonly available over-the-counter (St John's wort, echinacea, Ginkgo biloba, Asian ginseng, garlic). The information was searched for key safety messages identified by the National Center for Complementary and Integrative Health (NCCIH). We also explored the presence of risk of harm information. RESULTS: We recorded a rise in the number of products registered with the THR scheme (37% in 2016 compared to 7% in 2011). We also identified a reduction in the number of products that did not contain key safety information (75% in 2011 compared to 20% of products obtained in 2016). Risk of harm information was only communicated in products containing a PIL. We identified more products containing frequency of risk of harm information but this was not statistically significant. CONCLUSION: The introduction of the THR scheme appears to be associated with an increase in the provision of information about key safety messages on the safe use of herbal products. However, it is important to note that at least half of the products on the market that are not included in the THR scheme do not contain any information about their safe use; this includes information about precautions, interactions and side effects. The use of NCCIH herbal monographs replicated the methods used in the previous study; we recognise that the use of a different resource might effect the appraisal of the information provided. We also acknowledge that surveying presence of information does not assure that the latter is effectively communicated to patients, for which a close textual analysis would be required. While it is promising that more information is available after the introduction of the THR scheme, the public needs to be informed about ways to optimise safe use of all herbal products.

Characterization of cytokine gene expression in uterine cytobrush samples of non-endometritic versus endometritic postpartum dairy cows.

To better understand uterine inflammation in postpartum dairy cows we collected sequential cytobrush samples at 29-35 and at 49-55 d in milk (DIM). Based on the uterine cytology, cows were classified as Non-endometritic (n = 23; <18% neutrophils) or Endometritic (n = 12; ≥18% neutrophils) at 29-35 DIM and Non-endometritic (n = 17; <10% neutrophils) or Endometritic (n = 9; ≥10% neutrophils) at 49-55 DIM. Cows defined as Sham Controls (n = 6) were examined by vaginoscopy at 29-35 DIM and identified as Non-endometritic (<10% neutrophils) at 49-55 DIM. Cytokine gene expression in cytobrush samples was assessed using qRT-PCR. Sham Controls did not differ significantly (P > 0.17) from Non-endometritic cows at 49-55 DIM and these data were combined (n = 23). Uterine cytology-based classification using the aforementioned thresholds effectively separated cows into groups with Endometritic cows having significantly higher expression of pro-inflammatory (interleukin (IL)-1α, IL-1ß, IL-6, IL-8, IL-17A CSF-1; P < 0.01) and regulatory (IL-1RA and IL-10; P < 0.03) cytokines, relative to Non-endometritic cows. Furthermore, Non-endometritic cows showed a significant decline (P < 0.03) in the expression of pro-inflammatory (IL-1α, IL-6, IL-8) and regulatory (IL-10) cytokine genes as the postpartum period progressed; whereas Endometritic cows exhibited a sustained elevation in transcript abundance throughout the sample period for both pro-inflammatory and regulatory cytokine genes. Expression of transforming growth factor (TGF) genes was more complex with TGF-ß3 expression significantly (P < 0.01) lower at 29-35 DIM and TGF-ß1 gene expression significantly (P < 0.03) increased at 49-55 DIM in Endometritic versus Non-endometritic cows. Expression of TGF-ß2 gene was 2.7-fold higher (P < 0.01) at 29-35 DIM in cows that remained Endometritic when compared to cows recovering by 49-55 DIM. Some Non-endometritic cows (n = 4) at 29-35 DIM were reclassified as Endometritic at 49-55 DIM. The sampling procedures at 29-35 DIM did not alter either the cellular response (P > 0.43) or cytokine gene expression (P > 0.17) at 49-55 DIM. In conclusion, normal uterine involution is characterized by a progressive decline in pro-inflammatory and regulatory cytokine gene expression, while cows with endometritis show a dysregulated inflammatory process characterized by a sustained elevation in pro-inflammatory and regulatory cytokine gene expression. This analysis also shows that decreased TGF-ß2 gene expression at 29-35 DIM may be an indicator of recovery from endometritis.

Hypoxia determines survival outcomes of bacterial infection through HIF-1alpha dependent re-programming of leukocyte metabolism.

Hypoxia and bacterial infection frequently co-exist, in both acute and chronic clinical settings, and typically result in adverse clinical outcomes. To ameliorate this morbidity, we investigated the interaction between hypoxia and the host response. In the context of acute hypoxia, both S. aureus and S. pneumoniae infections rapidly induced progressive neutrophil mediated morbidity and mortality, with associated hypothermia and cardiovascular compromise. Preconditioning animals through longer exposures to hypoxia, prior to infection, prevented these pathophysiological responses and profoundly dampened the transcriptome of circulating leukocytes. Specifically, perturbation of HIF pathway and glycolysis genes by hypoxic preconditioning was associated with reduced leukocyte glucose utilisation, resulting in systemic rescue from a global negative energy state and myocardial protection. Thus we demonstrate that hypoxia preconditions the innate immune response and determines survival outcomes following bacterial infection through suppression of HIF-1α and neutrophil metabolism. The therapeutic implications of this work are that in the context of systemic or tissue hypoxia therapies that target the host response could improve infection associated morbidity and mortality.

An automated walk-over weighing system as a tool for measuring liveweight change in lactating dairy cows.

Automated walk-over weighing systems can be used to monitor liveweights of cattle. Minimal literature exists to describe agreement between automated and static scales, and no known studies describe repeatability when used for daily measurements of dairy cows. This study establishes the repeatability of an automated walk-over cattle-weighing system, and agreement with static electronic scales, when used in a commercial dairy herd to weigh lactating cows. Forty-six lactating dairy cows from a seasonal calving, pasture-based dairy herd in southwest Victoria, Australia, were weighed once using a set of static scales and repeatedly using an automated walk-over weighing system at the exit of a rotary dairy. Substantial agreement was observed between the automated and static scales when assessed using Lin's concordance correlation coefficient. Weights measured by the automated walkover scales were within 5% of those measured by the static scales in 96% of weighings. Bland and Altman's 95% limits of agreement were -23.3 to 43.6 kg, a range of 66.9 kg. The 95% repeatability coefficient for automated weighings was 46.3 kg. Removal of a single outlier from the data set increased Lin's concordance coefficient, narrowed Bland and Altman's 95% limits of agreement to a range of 32.5 kg, and reduced the 95% repeatability coefficient to 18.7 kg. Cow misbehavior during walk-over weighing accounted for many of the larger weight discrepancies. The automated walk-over weighing system showed substantial agreement with the static scales when assessed using Lin's concordance correlation coefficient. This contrasted with limited agreement when assessed using Bland and Altman's method, largely due to poor repeatability. This suggests the automated weighing system is inadequate for detecting small liveweight differences in individual cows based on comparisons of single weights. Misbehaviors and other factors can result in the recording of spurious values on walk-over scales. Excluding outlier weights and comparing means of 7 consecutive daily weights may improve agreement sufficiently to allow meaningful assessment of small short-term changes in automated weights in individuals and groups of cows.

SU-E-I-34: Air Kerma to Estimated Entrance Skin Dose Look-Up Tables: A Tool to Improve Dose Awareness in the Angiography Suite.

PURPOSE: Although air kerma (AK) is displayed during a case, often it does not represent the entrance skin dose (ESD), which can be estimated. The purpose of this work is to develop and provide system-customized AK-to- ESD look-up tables (LUTs) for immediate reference so that physicians can better evaluate the likelihood of deterministic skin reactions to weigh the risk-versus-benefit of continuing high-dose procedures. METHODS: Four correction factors are applied to estimate ESD from AK: inverse square correction from the interventional reference point to the average entrance table position, backscatter factor, mean energy absorption coefficient ratio, and measured table attenuation. Correction factors are room and service specific; therefore, room-specific AK-to-ESD LUTs are posted for easy reference. LUTs also list corresponding tissue reactions and their approximate time-of-onset. Protocols can be established for nurse or technology staff to provide verbal AK dose notifications during the case at crucial skin reaction dose thresholds (e.g. 2Gy indicating possible skin erythema and 5Gy indicating potentially prolonged recovery or permanent skin damage). Patient follow-up protocols can be established if the estimated ESD exceeds a set trigger level (e.g. 5Gy). Staff and physicians surveys evaluate usefulness and impact of dose awareness by system users. RESULTS: Two surveys report feedback on LUTs from physicians and technologists with 14 years median experience (range: 3-24 years). Over three-quarters of all angiography system users identify the LUTs and verbal dose notifications as positively affecting the institution's 'Patient First' initiative and roughly one-half of the imaging system users indicate that the LUTs and site-specific dose trigger level improves the dose awareness of care providers. CONCLUSIONS: Our efforts have focused on educating care providers about the differences between displayed AK and the estimated ESD. LUTs provide physicians and staff an immediate reference for estimated ESD and the associated deterministic skin effects at specific dose levels.

Evaluation of droplet dispersion during non-invasive ventilation, oxygen therapy, nebuliser treatment and chest physiotherapy in clinical practice: implications for management of pandemic influenza and other airborne infections.

BACKGROUND: Influenza viruses are thought to be spread by droplets, but the role of aerosol dissemination is unclear and has not been assessed by previous studies. Oxygen therapy, nebulised medication and ventilatory support are treatments used in clinical practice to treat influenzal infection are thought to generate droplets or aerosols. OBJECTIVES: Evaluation of the characteristics of droplet/aerosol dispersion around delivery systems during non-invasive ventilation (NIV), oxygen therapy, nebuliser treatment and chest physiotherapy by measuring droplet size, geographical distribution of droplets, decay in droplets over time after the interventions were discontinued. METHODS: Three groups were studied: (1) normal controls, (2) subjects with coryzal symptoms and (3) adult patients with chronic lung disease who were admitted to hospital with an infective exacerbation. Each group received oxygen therapy, NIV using a vented mask system and a modified circuit with non-vented mask and exhalation filter, and nebulised saline. The patient group had a period of standardised chest physiotherapy treatment. Droplet counts in mean diameter size ranges from 0.3 to > 10 µm were measured with an counter placed adjacent to the face and at a 1-m distance from the subject/patient, at the height of the nose/mouth of an average health-care worker. RESULTS: NIV using a vented mask produced droplets in the large size range (> 10 µm) in patients (p = 0.042) and coryzal subjects (p = 0.044) compared with baseline values, but not in normal controls (p = 0.379), but this increase in large droplets was not seen using the NIV circuit modification. Chest physiotherapy produced droplets predominantly of > 10 µm (p = 0.003), which, as with NIV droplet count in the patients, had fallen significantly by 1 m. Oxygen therapy did not increase droplet count in any size range. Nebulised saline delivered droplets in the small- and medium-size aerosol/droplet range, but did not increase large-size droplet count. CONCLUSIONS: NIV and chest physiotherapy are droplet (not aerosol)-generating procedures, producing droplets of > 10 µm in size. Due to their large mass, most fall out on to local surfaces within 1 m. The only device producing an aerosol was the nebuliser and the output profile is consistent with nebuliser characteristics rather than dissemination of large droplets from patients. These findings suggest that health-care workers providing NIV and chest physiotherapy, working within 1 m of an infected patient should have a higher level of respiratory protection, but that infection control measures designed to limit aerosol spread may have less relevance for these procedures. These results may have infection control implications for other airborne infections, such as severe acute respiratory syndrome and tuberculosis, as well as for pandemic influenza infection.

Expression of the repulsive SLIT/ROBO pathway in the human endometrium and Fallopian tube.

We investigated whether the repulsive SLIT/ROBO pathway is expressed in the endometrium and is negatively regulated during implantation. We also examined whether deficient expression in the Fallopian tube (FT) may predispose to ectopic pregnancy (EP). Endometrium (n = 21) and FT (n = 17) were collected across the menstrual cycle from fertile women with regular cycles. Decidualized endometrium (n = 6) was obtained from women undergoing termination, and FT (n = 6) was obtained from women with EP. SLIT/ROBO expression was quantified by reverse transcription-PCR and protein localized by immunohistochemistry. The regulation of SLIT/ROBO expression in vitro, by sex steroids and hCG, was assessed in endometrial (hTERT-EEpC) epithelial cells, and the effects of Chlamydia trachomatis infection and smoking were studied in oviductal (OE-E6/E7) epithelial cells. Endometrial SLIT3 was highest in the mid-secretory phase (P = 0.0003) and SLIT1,2 and ROBO1 showed a similar trend. ROBO2 was highest in proliferative phase (P = 0.027) and ROBO3,4 showed a similar trend. SLIT2,3 and ROBO1, 4 were lower in decidua compared with mid-secretory endometrium (P < 0.05). SLITs and ROBOs, excepting ROBO2, were expressed in FT but there were no differences across the cycle or in EP. SLIT/ROBO proteins were localized to endometrial and FT epithelium. Treatment of hTERT-EEpC with a combination of estradiol and medroxyprogesterone acetate inhibited ROBO1 expression (P < 0.01) but hCG had no effect. Acute treatment of OE-E6/E7 with smoking metabolite, cotinine, and C. trachomatis had no effect. These findings imply a regulated role for the endometrial SLIT/ROBO interaction during normal development and pregnancy but that it may not be important in the aetiology of EP.

Care homes' use of medicines study: prevalence, causes and potential harm of medication errors in care homes for older people.

INTRODUCTION: Care home residents are at particular risk from medication errors, and our objective was to determine the prevalence and potential harm of prescribing, monitoring, dispensing and administration errors in UK care homes, and to identify their causes. METHODS: A prospective study of a random sample of residents within a purposive sample of homes in three areas. Errors were identified by patient interview, note review, observation of practice and examination of dispensed items. Causes were understood by observation and from theoretically framed interviews with home staff, doctors and pharmacists. Potential harm from errors was assessed by expert judgement. RESULTS: The 256 residents recruited in 55 homes were taking a mean of 8.0 medicines. One hundred and seventy-eight (69.5%) of residents had one or more errors. The mean number per resident was 1.9 errors. The mean potential harm from prescribing, monitoring, administration and dispensing errors was 2.6, 3.7, 2.1 and 2.0 (0 = no harm, 10 = death), respectively. Contributing factors from the 89 interviews included doctors who were not accessible, did not know the residents and lacked information in homes when prescribing; home staff's high workload, lack of medicines training and drug round interruptions; lack of team work among home, practice and pharmacy; inefficient ordering systems; inaccurate medicine records and prevalence of verbal communication; and difficult to fill (and check) medication administration systems. CONCLUSIONS: That two thirds of residents were exposed to one or more medication errors is of concern. The will to improve exists, but there is a lack of overall responsibility. Action is required from all concerned.

A random phased array device for delivery of high intensity focused ultrasound.

Randomized phased arrays can offer electronic steering of a single focus and simultaneous multiple foci concomitant with low levels of secondary maxima and are potentially useful as sources of high intensity focused ultrasound (HIFU). This work describes laboratory testing of a 1 MHz random phased array consisting of 254 elements on a spherical shell of radius of curvature 130 mm and diameter 170 mm. Acoustic output power and efficiency are measured for a range of input electrical powers, and field distributions for various single- and multiple-focus conditions are evaluated by a novel technique using an infrared camera to provide rapid imaging of temperature changes on the surface of an absorbing target. Experimental results show that the array can steer a single focus laterally to at least +/-15 mm off axis and axially to more than +/-15 mm from the centre of curvature of the array and patterns of four and five simultaneous foci +/-10 mm laterally and axially whilst maintaining low intensity levels in secondary maxima away from the targeted area in good agreement with linear theoretical predictions. Experiments in which pork meat was thermally ablated indicate that contiguous lesions several cm(3) in volume can be produced using the patterns of multiple foci.

Clinical trial: oral prednisolone metasulfobenzoate (Predocol) vs. oral prednisolone for active ulcerative colitis.

BACKGROUND: Systemic corticosteroids are effective in ulcerative colitis but commonly cause side effects. AIM: To compare the safety and efficacy of a sparingly absorbed formulation of prednisolone metasulfobenzoate (Predocol) with a conventional tapering course of oral prednisolone. METHOD: In a double-blind randomized study, 59 active ulcerative colitis patients received Predocol 40 mg/day for 6 months, 61 received Predocol 60 mg/day for 6 months and 61 received prednisolone 40 mg/day for 2 weeks, tapered to week 8, followed by placebo until 6 months. RESULTS: Steroid-related side effects assessed using a 10-cm visual analogue scale were fewer at 2 months with Predocol 40 mg [VAS 8.1 cm (2.6), mean (s.d.)], or 60 mg [8.1 (2.1)] compared with prednisolone [6.7 (2.7); P = 0.01]. Mood changes affected 43% receiving prednisolone at 4 weeks vs. 8% for Predocol 40 mg (P = 0.001). Remission rates (Powell-Tuck < or =2) at 2 months were Predocol 40 mg 46%, Predocol 60 mg 28% and tapering prednisolone 41% (P = 0.13). Visual analogue scale for efficacy also showed non-inferiority for Predocol 40 mg/day. Remission rates at 6 months were Predocol 40 mg 51%, Predocol 60 mg 38% and tapering prednisolone 32% (P = 0.08). CONCLUSION: Predocol 40 mg/day has similar efficacy but markedly fewer side effects than a conventional tapering prednisolone regimen (ISRCTN14133410).

Epigenetic regulation of the ras effector/tumour suppressor RASSF2 in breast and lung cancer.

RASSF2 is a recently identified member of a class of novel tumour suppressor genes, all containing a ras-association domain. RASSF2 resides at 20p13, a region frequently lost in human cancers. In this report we investigated methylation status of the RASSF2 promoter CpG island in a series of breast, ovarian and non-small cell lung cancers (NSCLC). RASSF2 was frequently methylated in breast tumour cell lines (65%, 13/20) and in primary breast tumours (38%, 15/40). RASSF2 expression could be switched back on in methylated breast tumour cell lines after treatment with 5'-aza-2'deoxycytidine. RASSF2 was also frequently methylated in NSCLC tumours (44%, (22/50). The small number of corresponding normal breast and lung tissue DNA samples analysed were unmethylated. We also did not detect RASSF2 methylation in ovarian tumours (0/17). Furthermore no mutations were found in the coding region of RASSF2 in these ovarian tumours. We identified a highly conserved putative bipartite nuclear localization signal (NLS) and demonstrated that endogenous RASSF2 localized to the nucleus. Mutation of the putative NLS abolished the nuclear localization. RASSF2 suppressed breast tumour cell growth in vitro and in vivo, while the ability of NLS-mutant RASSF2 to suppress growth was much diminished. Hence we demonstrate that RASSF2 has a functional NLS that is important for its tumour suppressor gene function. Our data from this and a previous report indicate that RASSF2 is frequently methylated in colorectal, breast and NSCLC tumours. We have identified RASSF2 as a novel methylation marker for multiple malignancies and it has the potential to be developed into a valuable marker for screening several cancers in parallel using promoter hypermethylation profiles.

Redox-mediated substrate recognition by Sdp1 defines a new group of tyrosine phosphatases.

Reactive oxygen species trigger cellular responses by activation of stress-responsive mitogen-activated protein kinase (MAPK) signalling pathways. Reversal of MAPK activation requires the transcriptional induction of specialized cysteine-based phosphatases that mediate MAPK dephosphorylation. Paradoxically, oxidative stresses generally inactivate cysteine-based phosphatases by thiol modification and thus could lead to sustained or uncontrolled MAPK activation. Here we describe how the stress-inducible MAPK phosphatase, Sdp1, presents an unusual solution to this apparent paradox by acquiring enhanced catalytic activity under oxidative conditions. Structural and biochemical evidence reveals that Sdp1 employs an intramolecular disulphide bridge and an invariant histidine side chain to selectively recognize a tyrosine-phosphorylated MAPK substrate. Optimal activity critically requires the disulphide bridge, and thus, to the best of our knowledge, Sdp1 is the first example of a cysteine-dependent phosphatase that couples oxidative stress with substrate recognition. We show that Sdp1, and its paralogue Msg5, have similar properties and belong to a new group of phosphatases unique to yeast and fungal taxa.

Measurement properties of the CHAMPS physical activity questionnaire in a sample of older Australians.

BACKGROUND: The effective evaluation of physical activity interventions for older adults requires measurement instruments with acceptable psychometric properties that are sufficiently sensitive to detect changes in this population. AIM: To assess the measurement properties (reliability and validity) of the Community Healthy Activities Model Program for Seniors (CHAMPS) questionnaire in a sample of older Australians. METHODS: CHAMPS data were collected from 167 older adults (mean age 79.1 S.D. 6.3 years) and validated with tests of physical ability and the SF-12 measures of physical and mental health. Responses from a sub-sample of 43 older adults were used to assess 1-week test-retest reliability. RESULTS: Approximately 25% of participants needed assistance to complete the CHAMPS questionnaire. There were low but significant correlations between the CHAMPS scores and the physical performance measures (rho=0.14-0.32) and the physical health scale of the SF-12 (rho=0.12-0.24). Reliability coefficients were highest for moderate-intensity (ICC=0.81-0.88) and lowest for vigorous-intensity physical activity (ICC=0.34-0.45). Agreement between test-retest estimates of sufficient physical activity for health benefits (> or =150min and > or =5 sessions per week) was high (percent agreement=88% and Cohen's kappa=0.68). CONCLUSION: These findings suggest that the CHAMPS questionnaire has acceptable measurement properties, and is therefore suitable for use among older Australian adults, as long as adequate assistance is provided during administration.

Echinacea alkamide disposition and pharmacokinetics in humans after tablet ingestion.

Echinacea is a widely used herbal remedy for the treatment of colds and other infections. However, almost nothing is known about the disposition and pharmacokinetics of any of its components, particularly the alkamides and caffeic acid conjugates which are thought to be the active phytochemicals. In this investigation, we have examined serial plasma samples from 9 healthy volunteers who ingested echinacea tablets manufactured from ethanolic liquid extracts of Echinacea angustifolia and Echinacea purpurea immediately after a standard high fat breakfast. Caffeic acid conjugates could not be identified in any plasma sample at any time after tablet ingestion. Alkamides were rapidly absorbed and were measurable in plasma 20 min after tablet ingestion and remained detectable for up to 12 h. Concentration-time curves for 2,4-diene and 2-ene alkamides were determined. The maximal concentrations for the sum of alkamides in human plasma were reached within 2.3 h post ingestion and averaged 336+/-131 ng eq/mL plasma. No obvious differences were observed in the pharmacokinetics of individual or total alkamides in 2 additional fasted subjects who took the same dose of the echinacea preparation. This single dose study provides evidence that alkamides are orally available and that their pharmacokinetics are in agreement with the one dose three times daily regimen already recommended for echinacea.

Effects of isoflurane and xenon on Ba2+-currents mediated by N-type calcium channels.

BACKGROUND: Isoflurane and xenon are inhalation general anaesthetics with differing clinical profiles and contrasting synaptic actions. Both agents have been shown to depress excitatory synaptic responses. Whether this is via pre-synaptic or post-synaptic mechanisms has not been determined clearly. N-type calcium channels are a putative pre-synaptic target for these agents. We tested whether N-type calcium channels were sensitive to isoflurane and xenon and whether there was any stereoselectivity in the effect of isoflurane. METHODS: We used patch-clamp electrophysiology on isolated HEK293 cells stably expressing N-type calcium channels to investigate the effects of isoflurane and xenon on barium currents mediated by N-type calcium channels. RESULTS: Racemic isoflurane caused a concentration-dependent reduction (11-35%) in the peak current through the N-type channels in the concentration range 0.15-1.22 mM. In the clinically relevant concentration range the inhibition was small. At an isoflurane concentration of 0.31 mM (equivalent to 1 MAC), the peak N-type current was inhibited by 14 (1)%. The optical isomers of isoflurane were found to be equally potent at inhibiting currents through N-type channels. The inert gas anaesthetic xenon was found to have no measureable effect on N-type channels at a concentration of 3.4 mM (approximately 1 MAC). CONCLUSIONS: These results suggest that N-type calcium channels are not the targets mediating general anaesthesia with these two inhalation agents.

Epigenetic inactivation of SLIT3 and SLIT1 genes in human cancers.

In Drosophila, the Slit gene product, a secreted glycoprotein, acts as a midline repellent to guide axonal development during embryogenesis. Three human Slit gene orthologues have been characterised and recently we reported frequent promoter region hypermethylation and transcriptional silencing of SLIT2 in lung, breast, colorectal and glioma cell lines and primary tumours. Furthermore, re-expression of SLIT2 inhibited the growth of cancer cell lines so that SLIT2 appears to function as a novel tumour suppressor gene (TSG). We analysed the expression of SLIT3 (5q35-34) and SLIT1 (1q23.3-q24) genes in 20 normal human tissues. Similar to SLIT2 expression profile, SLIT3 is expressed strongly in many tissues, while SLIT1 expression is neuronal specific. We analysed the 5' CpG island of SLIT3 and SLIT1 genes in tumour cell lines and primary tumours for hypermethylation. SLIT3 was found to be methylated in 12 out of 29 (41%) of breast, one out of 15 (6.7%) lung, two out of six (33%) colorectal and in two out of (29%) glioma tumour cell lines. In tumour cell lines, silenced SLIT3 associated with hypermethylation and was re-expressed after treatment with 5-aza-2'-deoxycytidine. In primary tumours, SLIT3 was methylated in 16% of primary breast tumours, 35% of gliomas and 38% of colorectal tumours. Direct sequencing of bisulphite-modified DNA from methylated tumour cell lines and primary tumours demonstrated that majority of the CpG sites analysed were heavily methylated. Thus, both SLIT2 and SLIT3 are frequently methylated in gliomas and colorectal cancers, but the frequency of SLIT3 methylation in lung and breast cancer is significantly less than that for SLIT2. We also demonstrated SLIT1 promoter region hypermethylation in glioma tumour lines (five out of six; 83%), the methylation frequency in glioma tumours was much lower (two out of 20; 10%). Hence, evidence is accumulating for the involvement of members of the guidance cues molecules and their receptors in tumour development.