Left dorsolateral prefrontal cortex supports context-dependent prioritisation of off-task thought.

When environments lack compelling goals, humans often let their minds wander to thoughts with greater personal relevance; however, we currently do not understand how this context-dependent prioritisation process operates. Dorsolateral prefrontal cortex (DLPFC) maintains goal representations in a context-dependent manner. Here, we show this region is involved in prioritising off-task thought in an analogous way. In a whole brain analysis we established that neural activity in DLPFC is high both when 'on-task' under demanding conditions and 'off-task' in a non-demanding task. Furthermore, individuals who increase off-task thought when external demands decrease, show lower correlation between neural signals linked to external tasks and lateral regions of the DMN within DLPFC, as well as less cortical grey matter in regions sensitive to these external task relevant signals. We conclude humans prioritise daydreaming when environmental demands decrease by aligning cognition with their personal goals using DLPFC.

Glucose and the wandering mind: not paying attention or simply out of fuel?

RATIONALE/OBJECTIVES: The impact of raising glycaemia by ingestion of a glucose drink has revealed cognitive facilitation, particularly for memory and attention. This study aimed to extend current knowledge by examining, for the first time, whether glucose load also moderates task-related (TRT) and task-unrelated thoughts (TUT) during activities that vary in their requirement for sustained attention. METHOD: A 2 (25 g glucose vs. placebo) × 2 (fast vs. slow version of the Sustained Attention to Response Task (SART)) repeated measures, counterbalanced design was used with 16 healthy adults. Self-report questionnaires probed participants' levels of TRT and TUT during SART performance. Prior to testing, the Short Imaginal Processes Inventory (SIPI) was also administered to help pinpoint the nature of thought processes during the task before and after treatment. RESULTS: Analysis of variance revealed no significant effect of treatment; however, we report a pattern of results that is consistent with glucose facilitation effects on task accuracy for more demanding attention tasks (d = 0.56). Additionally, glucose improved the monitoring and task reflection as measured by TRT (d = 0.33) in the more demanding task but no effect on TUT. Probing the nature of thought processes further, we also report two novel correlations (in the placebo) between fears of failure (indexed by the SIPI) and the number of TUT episodes and perceived poor attention control (indexed by the SIPI) and number of TUT and speculate that glucose may act to buffer against TUT episodes under externally demanding situations. CONCLUSIONS: These data extend previous research examining the glucose facilitation effect to the processing of internal thought processes.

Two anatomic resources of canine pelvic limb muscles based on CT and MRI.

Advances in magnetic resonance (MR) imaging and three-dimensional (3D) modeling software provide the tools necessary to create sophisticated, interactive anatomic resources that can assist in the interpretation of MR images of extremities, and learning the structure and function of limb musculature. Modeling provides advantages over dissection or consultation of print atlases because of the associated speed, flexibility, 3D nature, and elimination of superimposed arrows and labels. Our goals were to create a diagnostic atlas of pelvic limb muscles that will facilitate interpretation of MR images of patients with muscle injury and to create a 3D model of the canine pelvic limb musculature to facilitate anatomic learning. To create these resources, we used structural segmentation of MR images, a process that groups image pixels into anatomically meaningful regions. The Diagnostic Atlas is an interactive, multiplanar, web-based MR atlas of the canine pelvic limb musculature that was created by manually segmenting clinically analogous MR sequences. Higher resolution volumetric MR and computed tomography (CT) data were segmented into separately labeled volumes of data and then transformed into a multilayered 3D computer model. The 3D Model serves as a resource for students of gross anatomy, encouraging integrative learning with its highly interactive and selective display capabilities. For clinicians, the 3D Model also serves to bridge the gap between topographic and tomographic anatomy, displaying both formats alongside, or even superimposed over each other. Both projects are hosted on an open-access website, http://3dvetanatomy.ncsu.edu/

Early experience with laparoscopic extralevator abdominoperineal excision within an enhanced recovery setting: analysis of short-term outcomes and quality of life.

INTRODUCTION: Conventional abdominoperineal excision for low rectal cancer has a higher local recurrence and reduced survival compared to anterior resection. An extralevator abdominoperineal excision (ELAPE) may improve outcome through removal of increased tissue in the distal rectum. Experience with ELAPE is limited and no studies have reported on quality of life (QOL) following this procedure. We describe a minimally invasive approach to ELAPE within an enhanced recovery programme, and present short-term results and QOL analyses. METHODS: All laparoscopic ELAPEs were included in a prospective database. Demographics, intra-operative and post-operative outcomes were evaluated. Postoperative QOL was assessed using the European Organisation for Research and Treatment of Cancer (EORTC) questionnaires QLQ-C30 and QLQ-CR29. RESULTS: Thirteen laparoscopic ELAPEs were performed over a two-year period. All were enrolled in an enhanced recovery programme. The median age was 76. The median tumour height was 20 mm (range: 0-50 mm) from the dentate line and all patients received neoadjuvant treatment. The median duration of surgery was 300 minutes (range: 120-488 minutes), the mean blood loss was 150 ml and one procedure was converted to open surgery. There was no circumferential resection margin involvement or tumour perforation. The median duration of use of intravenous fluid, patient controlled analgesia and urinary catheterisation was 2, 2 and 2.5 days respectively and the median length of hospital stay was 7.5 days. Two patients developed perineal wound dehiscence. QOL analysis revealed high global health status (90.8), physical (91.3), emotional (98.3) and social functioning (100) scores, which compared favourably with EORTC reference values and published QOL scores following conventional abdominoperineal excision. CONCLUSIONS: Laparoscopic ELAPE within an enhanced recovery setting is a feasible and safe approach with acceptable short-term outcomes and post-operative quality of life.

Melanoma sentinel node biopsy and prediction models for relapse and overall survival.

BACKGROUND: To optimise predictive models for sentinal node biopsy (SNB) positivity, relapse and survival, using clinico-pathological characteristics and osteopontin gene expression in primary melanomas. METHODS: A comparison of the clinico-pathological characteristics of SNB positive and negative cases was carried out in 561 melanoma patients. In 199 patients, gene expression in formalin-fixed primary tumours was studied using Illumina's DASL assay. A cross validation approach was used to test prognostic predictive models and receiver operating characteristic curves were produced. RESULTS: Independent predictors of SNB positivity were Breslow thickness, mitotic count and tumour site. Osteopontin expression best predicted SNB positivity (P=2.4 × 10⁻7), remaining significant in multivariable analysis. Osteopontin expression, combined with thickness, mitotic count and site, gave the best area under the curve (AUC) to predict SNB positivity (72.6%). Independent predictors of relapse-free survival were SNB status, thickness, site, ulceration and vessel invasion, whereas only SNB status and thickness predicted overall survival. Using clinico-pathological features (thickness, mitotic count, ulceration, vessel invasion, site, age and sex) gave a better AUC to predict relapse (71.0%) and survival (70.0%) than SNB status alone (57.0, 55.0%). In patients with gene expression data, the SNB status combined with the clinico-pathological features produced the best prediction of relapse (72.7%) and survival (69.0%), which was not increased further with osteopontin expression (72.7, 68.0%). CONCLUSION: Use of these models should be tested in other data sets in order to improve predictive and prognostic data for patients.

The effects of glucose ingestion and glucose regulation on memory performance in older adults with mild cognitive impairment.

BACKGROUND/OBJECTIVES: Previous research investigating the impact of glucose ingestion and/or improvements in glucose regulation has found selective cognitive facilitation on episodic memory tasks in successful ageing and dementia. The present study aimed to extend this research to mild cognitive impairment (MCI). SUBJECTS/METHODS: In a repeated-measures design, 24 older adults with and 24 older adults without MCI performed a battery of memory and attention tasks after 25 g of glucose or a sweetness matched placebo. In addition, to assess the impact of individual differences in glucose regulation, blood glucose measurements were taken throughout the testing session. RESULTS: Consistent with previous research, cognitive facilitation was observed for episodic memory tasks only in both successful ageing and MCI. Older adults with MCI had a similar glucose regulatory response as controls but their fasting levels were elevated. Notably, higher levels of blood glucose were associated with impaired memory performance in both the glucose and placebo conditions. Importantly, both blood glucose and memory performance indices were significant predictors of MCI status. CONCLUSIONS: The utility of glucose supplementation and the use of glucose regulation as a biological marker are discussed in relation to these data.

Spatial and temporal epidemiology of sporadic human cases of Escherichia coli O157 in Scotland, 1996-1999.

In Scotland, between 1995 and 2000 there were between 4 and 10 cases of illness per 100000 population per year identified as being caused by Escherichia coli O157, whereas in England and Wales there were between 1 and 2 cases per 100000 population per year. Within Scotland there is significant regional variation. A cluster of high rate areas was identified in the Northeast of Scotland and a cluster of low rate areas in central-west Scotland. Temporal trends follow a seasonal pattern whilst spatial effects appeared to be distant rather than local. The best-fit model identified a significant spatial trend with case rate increasing from West to East, and from South to North. No statistically significant spatial interaction term was found. In the models fitted, the cattle population density, the human population density, and the number of cattle per person were variously significant. The findings suggest that rural/urban exposures are important in sporadic infections.

Aromatherapy and behaviour disturbances in dementia: a randomized controlled trial.

A random controlled trial of the relaxing effects of an aromatherapy massage on disordered behaviour in dementia was conducted. Twenty-one patients were randomly allocated into one of three conditions, aromatherapy and massage (AM), conversation and aromatherapy (CA) and massage only (M). AM showed the greatest reduction in the frequency of excessive motor behaviour of all three conditions. This reached statistical significance between the hours of three and four pm (p < 0.05). Post hoc analysis suggested that at this time of day the AM consistently reduced motor behaviour when compared with CA (p = 0.05). This provides preliminary evidence of a measurable sedative effect of aromatherapy massage on dementia within a robust scientific paradigm. Further research is recommended with an expanded sample size.

Psychometric evaluation of a short observational tool for small-scale research projects in dementia.

Dementia is a degenerating illness and the lack of a reliable measure of self-report in particular presents particular difficulties for research. Often in the later stages of dementia behavioural measurement is the only tool available for the evaluation of treatment techniques. This paper describes and evaluates a short observational tool suitable for clinical assessment purposes. The scale has been shown to have the potential for adequate inter-rater reliability, test retest reliability, and convergent and divergent validity, if the study limitations reflecting statistical rather than ecological validity, and limitations of sample size are borne in mind.

Strangulating obstruction caused by intestinal herniation through the proximal aspect of the cecocolic fold in 9 horses.

OBJECTIVE: To report the clinical and surgical findings and outcome for horses with strangulating obstruction caused by herniation through the proximal aspect of the cecocolic fold. STUDY DESIGN: Retrospective study. ANIMALS: Nine horses. METHODS: Medical records were reviewed for clinical signs, surgical findings and technique, and outcome. Cadaver ponies and necropsy specimens were also used to study the regional anatomy of the cecocolic fold. RESULTS: The ileum and distal jejunum were strangulated in 8 horses, whereas in 1 horse the small intestine and the left ascending colons were incarcerated in a rent in the cecocolic fold. Two horses were euthanatized at surgery, 6 horses had a small intestinal resection (mean length, 3 m; range, 1.5-6.4 m) and an end-to-side jejunocecostomy, and the entrapment was reduced without resection in the horse that had small intestine and ascending colon incarceration; cecocolic fold defects were not closed. One horse was euthanatized 36 hours after surgery because of endotoxemia. Six horses were discharged; 4 were available for long-term follow-up, of which 2 were euthanatized, and 2 were euthanatized 12 and 18 months after surgery because of colic signs. Variations in thickness of the cecocolic fold were observed in specimens obtained from necropsy of other horses and ponies. CONCLUSIONS: Reasons for this defect are unknown, although observed anatomic differences in cecocolic fold thickness may contribute to the development of defects. CLINICAL RELEVANCE: Reduction of the entrapped bowel is easiest when traction is placed on the bowel at a 90 degrees to the base of the cecum. Intestinal incarceration through rents within the proximal part of the cecocolic fold should be considered as a differential diagnosis for strangulating obstruction in horses.

CD40 ligation for immunotherapy of solid tumours.

Tumour vaccines provide an important focus of current cancer research and are often based on the premise that although T-cells do respond naturally to certain tumours, this is usually weak and therefore ineffective at controlling disease. An integral and necessary part of a T-cell immune response involves triggering of CD40 on antigen-presenting cells (APC) by its ligand, CD154, on responding T helper (Th) cells. Furthermore, cytotoxic responses to tumours may fail because the Th-cell response is inadequate and unable to provide CD40 stimulation of APC. Growing evidence shows that stimulating APC with soluble CD40L or an agonistic anti-CD40 mAb can, at least in part, replace the need for Th cells and generate APC that are capable of priming cytotoxic T lymphocytes (CTL). The aim of this study was to investigate whether a range of solid tumours (CD40(-)) could be treated with anti-CD40 mAb. It was found that this treatment was effective, and correlated with the intrinsic immunogenicity and aggressiveness of the tumours. The mAb could be delivered locally or at a distal site, but increased antigen load provided by irradiated tumour cells added little to the effectiveness of the treatment. T-cells were required since cytokine (interferon-gamma) and CTL activity were demonstrated following treatment and the therapeutic efficacy was lost in nude mice. In addition, depletion of CD8(+) cells abrogated protection whilst depletion of CD4(+) cells had no effect. This study demonstrates that solid CD40(-) tumours are sensitive to anti-CD40 mAb therapy and that the response bypasses the need for Th cells.

Analysis of the PKC-gamma-related immunocrossreactive region of a novel leukocyte protein gamma-rp.

The newly discovered gamma-PKC-related-protein of human leukocytes (gamma-rp) crossreacts with a polyclonal antibody preparation originally designed to be specific for PKC-gamma (gammaMb-Ab). As this antibody is currently the only suitable probe for gamma-rp, we sought to characterize the binding of the two proteins. We determined that the gamma Mg-Ab does not recognize the native form of gamma-rp. However, with denaturing immunoblots of gamma-rp, we found that 1) the crossreactive gamma-rp epitope differs somewhat from that of classic rat brain PKC-gamma, but probably only to the degree of the rat/human PKC species difference; 2) the previously reported doublet bands of gamma-rp represent a single protein with cell-stimulus inducible modifications; 3) antibodies present in the gammaMg-Ab pool bind to two separate sites within the gamma-rp epitope; 4) access to one binding site is conformationally restricted, even after protein denaturation; 5) agonist-induced modification of gamma-rp does not significantly affect the total amount of gamma Mg-Ab that it can bind, but 6) does significantly affect the rate of antibody binding to one site. This investigation defines the appropriate experimental use of our antibody, and the significance of these findings for the future study and cloning of gamma-rp is discussed.

TECHNICAL NOTE: measurement of cAMP-dependent protein kinase activity using a fluorescent-labeled kemptide.

BACKGROUND: Traditional protein kinase assays include the use of [32P] labeled ATP as phosphate donor and a substrate protein or peptide as phosphoreceptor. Since this approach has a number of drawbacks in addition to generating ionizing radiation, several non-isotopic methods have been developed. Although shown to reflect the activity of purified enzymes, none have been demonstrated to detect physiological changes in endogenous enzyme activity in cell homogenates. METHODS: Studies were performed to examine the kinetics, reproducibility, and optimal assay conditions of a novel non-radioisotopic kinase assay that detects PKA activity by phosphorylation of the peptide substrate Kemptide covalently bound to a fluorescent molecule (f-Kemptide). Basal and agonist-induced PKA activity in epithelial cell homogenates was measured. RESULTS: The kinetics of f-Kemptide were similar to the standard radioisotopic method with intraassay and interassay variations of 5.6 +/- 0.8% and 14.3 +/- 2.6%, respectively. Neither fluorescence quenching nor enhancing effects were found with consistent amounts of homogenate protein. Specific PKA activity was determined as the IP20-inhibitable fraction to account for nonspecific phosphorylation, perhaps due to S6 kinase or a similar enzyme. The basal activity of 38% of total PKA in A6 cells increased by 84% after exposure to vasopressin and by 58% after short exposure to forskolin. In T84 cells exposed to VIP there was a 360% increase over basal activity. CONCLUSIONS: These results show that f-Kemptide exhibits acceptable kinetics, and that the assay system can quantitatively and reproducibly measure basal and stimulated PKA activity in cell homogenates.

Vasopressin-induced activation of protein kinase C in renal epithelial cells.

Recent studies indicate that the actions of arginine vasopressin (AVP) and other agonists that stimulate electrogenic sodium transport in renal epithelial A6 cells are linked to a Ca(2+)-mobilizing signal transduction mechanism that involves generation of inositol trisphosphate. Since diacylglycerol is the other product in this pathway, studies were performed to determine the possible role of PKC in the stimulation of sodium transport. AVP induced a biphasic increase in diacylglycerol generation, characterized by an initial rapid rise and then a sustained elevation, and PKC activation, reflected by phosphorylation of a specific 80 kDa myristoylated alanine-rich PKC substrate (MARCKS). To determine the PKC isoform(s) involved in this process, immunoblot analysis was performed using antisera that recognize both classical PKC isoforms, XPKC-I and XPCK-II, cloned from Xenopus oocytes. The transcripts of both isoforms were expressed in the A6 cell. Since protein recognized by antisera was translocated from cytosol to the particulate fraction after exposure to AVP, one or both isoforms were activated in the A6 cell. Further studies showed that cyclohexyladenosine and insulin, additional agonists of sodium transport in A6 cells, also stimulated phosphorylation of MARCKS. These results argue that Ca(2+)-dependent PKC is involved in the action of AVP, and that of other agonists, which stimulate sodium transport.

An apparently novel protein of human leukocytes, reactive with an antibody to protein kinase C-gamma, is rapidly modified upon cell activation: initial characterization in neutrophils and their cytoplasts.

On immunoblots of human neutrophil cytoplasts (U-CYT), a previously undescribed 97 kDa protein was revealed by intense and selective reaction with an antibody that was initially raised to recognize PKC-gamma. Denoted "gamma-rp" for gamma-related protein, this acidic cytosolic protein somewhat resembled the classic forms of PKC in several biochemical respects. Appearing as a doublet on low-percentage SDS-PAGE gels, both its mobility and staining pattern were rapidly altered by treatment of U-CYT with either phorbol ester or chemotactic peptide. Whole neutrophil gamma-rp was detectable only after TCA precipitation of intact cells. It was also detectable in human platelets, lymphocytes, and neutrophil-like differentiated HL60 cells, but not in fibroblasts, erythrocytes, monocytes, or monocyte-like differentiated HL60 cells. Our data suggest that gamma-rp merits further study as a potential participant in cellular activation, and as a possible structural or functional relative of PKC.

An ospA frame shift, identified from DNA in Lyme arthritis synovial fluid, results in an outer surface protein A that does not bind protective antibodies.

Passive immunization with murine or human Abs to outer surface protein A (OspA) can protect mice against Borrelia burgdorferi, but OspA Abs elicited during natural infection in mice or humans are unable to clear the spirochete from the infected host. To examine Ab binding by OspA during the course of human infection, we amplified the operon encoding full-length ospA and ospB from synovial fluids of a patient with chronic Lyme arthritis, the first such recoveries from human material, at four separate time points over 4.5 mo, and expressed OspA in Escherichia coli. OspA mAbs that passively protected mice from infection did not bind one of the expressed OspAs, because of a deletion in ospA that resulted in a frame shift and premature stop codon near the carboxyl terminus. However, expressed OspA from a later synovial fluid sample did not contain this deletion. Thus, although altered forms of OspA, which potentially can influence host immune effectiveness, do occur in the human host, they cannot be the only factors responsible for microbial persistence.

Vasopressin-stimulated electrogenic sodium transport in A6 cells is linked to a Ca(2+)-mobilizing signal mechanism.

Vasopressin is known to activate two types of cell surface receptors; V2, coupled to adenylate cyclase, and V1, linked to a Ca(2+)-dependent transduction system. We investigated whether arginine vasopressin (AVP) stimulation of electrogenic sodium transport in A6 cells, derived from Xenopus laevis, is mediated by activation of either one or both types of AVP-specific receptors. AVP caused a rapid increase in electrogenic sodium transport, reflected by the transepithelial potential difference (VT) and equivalent short circuit current (Ieq) measurements. AVP also rapidly increased intracellular Ca2+ (Ca2+i) and total inositol trisphosphate. The increase in Ieq was dependent on the rise in (Ca2+i), because 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) dose-dependently inhibited the Ieq response. There was no evidence, however, that activation of adenylate cyclase mediated AVP-stimulated Ieq; transport was not inhibited after AVP-induced activation of adenylate cyclase was abolished by 2',5'-dideoxyadenosine or when cAMP-dependent protein kinase (PKA) activity was abolished by the specific PKA inhibitor IP20. Further studies showed that although both forskolin and 8-(4-chlorophenylthio)-cAMP stimulated Ieq, this occurred by mechanisms independent of PKA activation. These results indicate that AVP-stimulated Na+ transport is mediated by a V1 receptor and a Ca(2+)-dependent mechanism.

Adenosine stimulation of Na+ transport is mediated by an A1 receptor and a [Ca2+]i-dependent mechanism.

Studies were performed to determine the primary signal transduction mechanism that mediates adenosine stimulation of electrogenic sodium transport in renal epithelial cells. Experiments were performed on cultured amphibian A6 cells with an adenosine analogue that preferentially binds to the A1 receptor, cyclohexyladenosine (CHA). Sodium transport was assessed by the equivalent short circuit current (Ieq). CHA was found to stimulate Ieq via activation of an A1 receptor because (1) the threshold concentration was 1 nM compared to that of 10 microM for the specific A2 agonist CGS21680, (2) CHA inhibited vasopressin (AVP)-stimulated cAMP production by a pertussis toxin-sensitive mechanism, and (3) the action of CHA was inhibited by the A1 antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX). CHA increased intracellular Ca2+ ([Ca2+]i) and stimulated phosphoinositide turnover at concentrations that increased Ieq and in a time course that paralleled the increase in Ieq. Ion transport was stimulated by a Ca(2+)-dependent mechanism because the CHA induced increase in Ieq was inhibited by chelating [Ca2+]i with 5,5'dimethyl BAPTA in a dose-dependent manner, with a Ki of approximately 10 microM. The increase in Ieq was also dose-dependently inhibited by the specific PKC inhibitors dihydroxychlorpromazine and chelerythrine, and by trifluoperazine which inhibits PKC and calmodulin. Further studies indicated that CHA-stimulated Ieq was independent of cAMP generation because CHA did not induce an increase in cAMP accumulation parallel to the increase in Ieq in a dose-response analysis, and the adenylate cyclase inhibitor 2',5' dideoxy-adenosine (DDA) did not affect the CHA-induced increase in Ieq.(ABSTRACT TRUNCATED AT 250 WORDS)

Misoprostol stimulates leukocyte cyclic adenosine 3',5' monophosphate production and synergizes with colchicine: novel combination of established drugs may boost anti-inflammatory potential.

Elevation of intracellular cyclic adenosine 3',5' monophosphate (cAMP) inhibits various proinflammatory and immune responses of leukocytes. Among agents known to stimulate cAMP production in these cells, prostaglandins E (PGEs) have received particular attention as potential immunosuppressive and/or anti-inflammatory drugs. Their clinical use, however, is limited by poor oral absorption and extreme metabolic instability. Misoprostol, a synthetic analog of PGE1 that can be given orally and that has a significantly longer biological half-life, is now used to prevent or treat nonsteroidal anti-inflammatory drug (NSAID)-induced gastric injury. Because it might also exert anti-inflammatory effects on leukocytes, we have characterized the effects of misoprostol on cAMP production in these cells. We have found that misoprostol does stimulate cAMP production, although with some-what less potency and maximal effect than PGE1; this stimulation is synergistically increased by pretreatment of cells with colchicine; a clinically relevant dose of colchicine is effective given sufficient pretreatment time, and preexposure of cells to colchicine enables a clinically relevant dose of misoprostol to stimulate cAMP generation. We conclude that colchicine and misoprostol represent a drug combination that might prove clinically useful for therapy of inflammatory disease.