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Atrial fibrillation (AF) is the most common significant cardiac rhythm disorder and is a powerful common risk factor for stroke. Randomized trials have demonstrated that anticoagulation can reduce the risk of stroke in patients with AF. Yet, there continues to be widespread underutilization of this therapy. To address this practice gap locally and improve efforts to reduce the risk of stroke for patients with AF in our health system, we have designed a study to implement and evaluate the effectiveness of an Atrial Fibrillation Decision Support Tool (AFDST) embedded within our electronic health record.Our intervention is provider-facing and focused on decision support. The clinical setting is ambulatory patients being seen by primary care physicians. Patients include those with both incident and prevalent AF. This randomized, prospective trial will enroll 800 patients in our University of Cincinnati Health System who are currently receiving less than optimal anticoagulation therapy as determined by the AFDST. Patients will be randomized to one of two arms - 1) usual care, in which the AFDST is available for use; 2) addition of a best practice advisory (BPA) to the AFDST notifying the clinician that their patient stands to gain a significant benefit from a change in their current thromboprophylactic therapy.The primary outcome is effectiveness of the BPA measured by change to "appropriate thromboprophylaxis" based on the AFDST recommendation at 3 months post randomization. Secondary endpoints include Reach and Adoption, from the RE-AIM framework for implementation studies. Sample size is based upon an improvement from inappropriate to appropriate anticoagulation therapy estimated at 4% in the usual care arm and ≥10% in the experimental arm.Our goal is to examine whether addition of a BPA to an AFDST focused on primary care physicians in an ambulatory care setting will improve "appropriate thromboprophylaxis" compared with usual care. Results will be examined at 3 months post randomization and at the end of the study to evaluate durability of changes. We expect to complete patient enrollment by the end of June 2022.Clinicaltrials.gov NCT04099485.

Appropriate thromboprophylaxis for patients with atrial fibrillation or atrial flutter (AF) remains a national challenge.We hypothesized that a shared decision-making interaction facilitated by an Atrial Fibrillation Shared Decision Making Tool (AFSDM) would improve patient knowledge about atrial fibrillation, and the risks and benefits of various treatment options for stroke prevention; increase satisfaction with the decision-making process; improve the therapeutic alliance between patient and the clinical care team; and increase medication adherence. Using a pre- and post-visit study design, we enrolled 76 patients and completed 2 office visits and 1-month telephone follow-up for 65 patients being seen in our Arrhythmia Clinic over the 1-year period (July 2016 through June 2017). Our primary outcome measure was change in decisional conflict between the first and second clinical visit.Decisional conflict decreased from an average of 31 to 9. Mean change was 22.3 (95% CI, 25.7 - 37.1), corresponding to an effect size of 0.94 standard deviations. Satisfaction with decision increased from 4.0 to 4.5, measures of therapeutic alliance with the care team (Kim Alliance scale) increased from 100.1 to 103.1, and satisfaction with provider increased from 4.2 to 4.5 (P.0001 for all measures). AF knowledge assessment scores increased from 8.4 to 9.1, and knowledge about personal stroke and bleeding risk increased from 1 to 1.5 (P.0001). Finally, medication adherence improved as reflected by an increase in the Morisky Medication Adherence scale from 5.9 to 6.4 (P.0001).A shared decision-making interaction, facilitated by an AFSDM can significantly improve multiple measures of decision-making quality, leading to improved medication adherence and patient satisfaction.

There has been increasing recognition that the alarming surge in allergy and autoimmunity in the industrialised and developing worlds shadows the rapid eradication of pathogens, such as parasitic helminths. Appreciation of this has fuelled an explosion in research investigating the therapeutic potential of these worms. This review considers the current state-of-play with a particular focus on exciting recent advances in the identification of potential novel targets for immunomodulation that can be exploited therapeutically. Furthermore, we contemplate the prospects for designing worm-derived immunotherapies for an ever-widening range of inflammatory diseases, including, for example, obesity, cardiovascular disease, and ageing as well as neurodevelopmental disorders like autism.

Background Appropriate thromboprophylaxis for patients with atrial fibrillation (AF) remains a national challenge. Methods We hypothesized that provision of decision support in the form of an Atrial Fibrillation Decision Support Tool (AFDST) would improve thromboprophylaxis for AF patients. We conducted a cluster randomized trial involving 15 primary care practices and 1,493 adults with nonvalvular AF in an integrated health care system between April 2014 and February 2015. Physicians in the intervention group received patient-level treatment recommendations made by the AFDST. Our primary outcome was the proportion of patients with antithrombotic therapy that was discordant from AFDST recommendation. Results Treatment was discordant in 42% of 801 patients in the intervention group. Physicians reviewed reports for 240 patients. Among these patients, thromboprophylaxis was discordant in 63%, decreasing to 59% 1 year later ( P = .02). In nonstratified analyses, changes in discordant care were not significantly different between the intervention group and control groups. In multivariate regression models, assignment to the intervention group resulted in a nonsignificant trend toward decreased discordance ( P = .29), and being a patient of a resident physician ( P = .02) and a higher HAS-BLED score predicted decreased discordance ( P = .03), whereas female gender ( P = .01) and a higher CHADSVASc score ( P = .10) predicted increased discordance. Conclusions Among patients whose physicians reviewed recommendations of the decision support tool discordant therapy decreased significantly over 1 year. However, in nonstratified analyses, the intervention did not result in significant improvements in discordant antithrombotic therapy.

Highlights • Small molecule analogues (SMAs) of the immunomodulator, ES-62, have been produced. • Two SMAs protect against oxazolone-induced skin inflammation in mouse ears. • Protection is associated with reduced cellular infiltration and collagen deposition. • Protection is associated with decreased IFNγ mRNA in the ears., Graphical Abstract, ES-62 is the major secreted protein of the rodent filarial nematode Acanthocheilonema viteae. The molecule contains covalently attached phosphorylcholine (PC) residues, which confer anti-inflammatory properties on ES-62, underpinning the idea that drugs based on this active moiety may have therapeutic potential in human diseases associated with aberrant inflammation. Here we demonstrate that two synthetic small molecule analogues (SMAs) of ES-62 termed SMA 11a and SMA 12b are protective in the oxazolone-induced acute allergic contact dermatitis mouse model of skin inflammation, as measured by a significant reduction in ear inflammation following their administration before oxazolone sensitisation and before oxazolone challenge. Furthermore, it was found that when tested, 12b was effective at reducing ear swelling even when first administered before challenge. Histological analysis of the ears showed elevated cellular infiltration and collagen deposition in oxazolone-treated mice both of which were reduced by treatment with the two SMAs. Likewise, the oxazolone-induced increase in IFNγ mRNA in the ears was reduced but no effect on other cytokines investigated was observed. Finally, no influence on the mast cell populations in the ear was observed.

Rheumatoid arthritis (RA) remains a debilitating autoimmune condition as many patients are refractory to existing conventional and biologic therapies, and hence successful development of novel treatments remains a critical requirement. Towards this, we now describe a synthetic drug-like small molecule analogue, SMA-12b, of an immunomodulatory parasitic worm product, ES-62, which acts both prophylactically and therapeutically against collagen-induced arthritis (CIA) in mice. Mechanistic analysis revealed that SMA-12b modifies the expression of a number of inflammatory response genes, particularly those associated with the inflammasome in mouse bone marrow-derived macrophages and indeed IL-1β was the most down-regulated gene. Consistent with this, IL-1β was significantly reduced in the joints of mice with CIA treated with SMA-12b. SMA-12b also increased the expression of a number of genes associated with anti-oxidant responses that are controlled by the transcription factor NRF2 and critically, was unable to inhibit expression of IL-1β by macrophages derived from the bone marrow of NRF2−/− mice. Collectively, these data suggest that SMA-12b could provide the basis of an entirely novel approach to fulfilling the urgent need for new treatments for RA., Graphical abstract, Highlights • SMA-12b is a mimetic of the active PC-moiety of the helminth immunomodulator, ES-62. • SMA-12b effectively protects mice against collagen-induced arthritis (CIA). • SMA-12b downregulates IL-1β and inflammasome genes via activation of NRF2. • SMA-12b reduces IL-1β in the joints of mice with CIA.

ES-62 is an anti-inflammatory phosphorylcholine-containing glycoprotein secreted by the filarial nematode Acanthocheilonema viteae. Accelerated atherosclerosis frequently occurs in systemic lupus erythematosus, resulting in substantial cardiovascular morbidity and mortality. We examined the effects of ES-62 in the gld.apoE(-/-) mouse model of this condition. Treatment with ES-62 did not substantially modulate renal pathology but caused decreased anti-nuclear autoantibody levels. Moreover, a striking 60% reduction in aortic atherosclerotic lesions was observed, with an associated decrease in macrophages and fibrosis. We believe that these latter findings constitute the first example of a defined parasitic worm product with therapeutic potential in atherosclerosis: ES-62-based drugs may represent a novel approach to control accelerated atherosclerosis in systemic lupus erythematosus.

The Moon spends 20% of its orbit within the terrestrial magnetosphere. During this time it experiences a dynamic plasma environment, including high-speed streams, flux ropes and a flux of heavy ions from ionospheric outflows. 3D multi-fluid simulations of the Moon within the magnetosphere during a substorm shows that a highly variable plasma flow can develop in the vicinity of the Moon due to the passage of a flux rope. The transit of a flux rope past the Moon potentially leads to a plasma wake that is mis-aligned from the optical wake by nearly 30 ° . This will have implications when determining the range of space weathering and surface charging the lunar surface experiences.

The Moon spends 20% of its orbit within the terrestrial magnetosphere. During this time it experiences a flux of ions from the Earth, which can be accelerated within the magnetosphere prior to impacting the Moon. Both 3D multi-fluid simulations and particle tracking were used to study the lunar environment while the Moon is inside the magnetosphere and quantify the energy and flux of ionospheric ions at lunar orbits for times ranging from moderate substorms to severe storms. The results from the fluid simulations indicate that during storms, the flux of ionospheric oxygen ions to the Moon can increase by two orders of magnitude and the bulk of those oxygen ions have energies up to 100s of keV. Particle tracking indicates that the peak energy of particles in the tail of the distribution approaches 1 MeV for the most intense storms, as particles from the ring current can be lost downtail to lunar orbits. Periods of increased flux of ionospheric particles intercepted by the Moon over geologic time periods would have played a role in weathering of the lunar surface and delivery of water components to the surface.

Negative regulation of B cell activation by cognate immune complexes plays an important homeostatic role in suppressing B cell hyperactivity and preventing consequent autoimmunity. Immune complexes co-ligate the BCR and FcγRIIB resulting in both growth arrest and apoptosis. We now show that such apoptotic signalling involves induction and activation of p53 and its target genes, the pro-apoptotic Bcl-2 family members, Bad and Bid, as well as nuclear export of p53. Collectively, these events result in destabilisation of the mitochondrial and lysosomal compartments with consequent activation and interplay of executioner caspases and endosomal-derived proteases. In addition, the upregulation of Fas and FasL with consequent activation of caspase 8-dependent death receptor signalling is required to facilitate efficient apoptosis of B cells. Consistent with this role for Fas death receptor signalling, apoptosis resulting from co-ligation of the BCR and FcγRIIB is defective in B cells from Fas-deficient MRL/MpJ-Fas(lpr) mice. As these mice develop spontaneous, immune complex-driven lupus-like glomerulonephritis, targeting this FcγRIIB-mediated apoptotic pathway may therefore have novel therapeutic implications for systemic autoimmune disease.

ES-62 is an immunomodulatory phosphorylcholine (PC)-containing glycoprotein secreted by the rodent filarial nematode Acanthocheilonema viteae. Previously, the use of knockout mice has revealed the effects of ES-62 on macrophages and dendritic cells to be dependent on TLR4. However, it is possible that ES-62 may interact with additional proteins on the surfaces of target cells and hence that cells may vary with respect to receptor usage. In this study, we identified by molecular weight, proteins that interact with ES-62 and found differences amongst the immune system cells studied. Thus, whereas lymphocytes appear to have two major interacting proteins of ∼135 and ∼82 kDa, U937 monocytes only contain an ES-62-binding protein of the latter molecular weight. Binding to the proteins on B cells and U937 cells was blocked by PC, suggesting a critical role for this ES-62 moiety in facilitating interaction. Finally, ES-62 binding is followed by internalization in both macrophages and B cells but only in the former was absence of TLR4 found to block internalization. These findings are consistent with differences in receptor usage by ES-62 amongst different cell-types.

INTRODUCTION The first scientific description of fluorescence was made by Sir George G. Stokes in 1852. Sir Stokes showed that fluorite (fluorspar) and uranium glass had fluorescent properties that enabled these compounds to convert UV light, invisible to the human eye, into one of a longer wavelength visible to the human eye. Light is emitted when an electron, previously lifted to a higher-energy state by excitation light, drops back to its lower ground state (Fig 1A). The difference in wavelength between the excitation and emission light is called the Stokes shift (Fig 1B) and forms the basis for fluorescencebased assays.

This study calculates the surface energy of polystyrene tissue culture plastic, silicon, silicon dioxide and indium tin oxide, all of which have applications in tissue culture. The adhesion molecules: collagen, fibronectin, poly-L-ornithine and poly-D-lysine, were coated onto these various surfaces, and the surface energy of the coated substrates calculated. Coating with fibronectin was found to produce a monopolar acidic surface while poly-D-lysine, poly-L-ornithine and collagen coatings were found to produce monopolar basic surfaces. The calculated surface energy components of the coated materials were then used to give a quantitative determination of the magnitude of their hydrophobicity. It was concluded that collagen, polylysine and polyornithine could provide a hydrophobic or hydrophilic surface depending on the underlying substrates they were coated on. The measurement obtained for fibronectin, unlike the other adhesion molecules, was independent of the underlying surface and remained hydrophobic on all substrates tested. Wetting experiments were carried out on the coated substrates, using the tissue culture medium Dulbeccos modified eagles medium, both containing and not containing serum proteins, and saline solution. These liquids that are commonly used in tissue culture, were then used to provide information how these liquids behave on various substrates coated with the adhesion molecules. Results show that fibronectin coated surfaces represent the most phobic surface for all three liquids. The findings of this study can be used in cell manipulation studies and provide a valuable data set for the biomedical and research industries.

Filarial nematodes persist in the parasitized host by modulating immune responsiveness. A feature of this that has been observed in a multitude of studies dating back several decades is an inability of lymphocytes to respond appropriately to filarial nematode antigens and, in some cases, to other stimuli. The consistency of this observation, allied to the ease of measurement of lymphocyte hyporesponsiveness, has resulted in many attempts to understand its cause.

The molecular mechanism by which the antigen receptors (BCR) on B cells can elicit differential maturation state-specific responses is one of the central problems in B-cell differentiation yet to be resolved. Indeed, many of the early signalling events detected following BCR ligation, such as activation of protein tyrosine kinases (PTK), phospholipase C (PLC), phosphoinositide-3-kinase (PI 3K), protein kinase C (PKC) and the RasMAPK (mitogen activating protein kinase) signalling cascades are observed throughout B-cell maturation. However, it is becoming clear that the differential functional responses of these BCR-coupled signals observed during B-cell maturation are dependent on a number of parameters including signal strength and duration, subcellular localisation of the signal, maturation-restricted expression of downstream signalling effector elements/isoforms and modulation of signal by co-receptors. Thus, the combined signature of BCR signalling is likely to dictate the functional response and act as a developmental checkpoint for B-cell maturation.

Modulation of immune responses is an important strategy employed by pathogens to enable their survival in host organisms. Secreted immunomodulatory molecules are key weapons in the pathogen's battle with the host immune system. In this review, we will discuss the immunomodulatory effects of the phosphorylcholine-containing filarial nematode glycoprotein, ES-62, on the host immune system and summarise the results of our studies to identify the intracellular signalling pathways targeted by ES-62 to achieve these effects.

Catastrophic events including those sparked by terrorism have cast a pall over society in recent years. Events such as September 11th or the bombing of the Alfred P. Murrah Federal Building in Okalahoma City demonstrated a sudden surge in telephone call volume. These events indicated that communications systems, including cellular, can fail as a result from damage or capacity overload. Cellular systems have significant capacity given the limited radio spectrum allocated but much like airlines that oversell seats because inevitably some passengers do not show up, cellular providers typically have more customers than operational spectrum can handle in aggregate. Normally this is perfectly acceptable because a high percentage of the customer base would never be making calls simultaneously; except in an emergency. Such emergencies can be active or passive in nature. The release of dangerous airborne agents is one such scenario. The purpose of this study was to use a portable air particulate sensor as a data acquisition platform to evaluate the capability of real-time data throughput using existing wireless topologies, Java, eXtensible Markup Language (XML) and Internet protocols when cellular transmission frequencies are compromised. Results show that high levels of cellular voice traffic in a given area can hamper simultaneous data communications; however, Low Earth Orbiting Satellites (LEOS) can provide redundancy.

Arthropod-transmitted filarial nematodes can survive for in excess of a decade via modulation of the vertebrate host immune system. Although human infection can result in very severe pathology, most infected individuals show remarkably little evidence of this. Analysis of the anti-nematode response indicates that apparently pathology-free individuals have an anti-inflammatory immunological phenotype and it has been suggested that this favours maintenance of host good health. It is considered that parasite-derived molecular secretions contribute to the anti-inflammatory phenotype and we have thus investigated the properties of a filarial nematode glycoprotein secreted in some abundance, ES-62. This molecule shows a plethora of immunomodulatory activities that can be classified as anti-inflammatory. It has been observed in a number of studies that several autoimmune disorders including rheumatoid arthritis (RA) exhibit reduced incidence and severity in geographic regions in which filarial nematodes are transmitted to humans. Furthermore, it has been speculated that these two observations are linked although molecular explanations for such an association have not been forthcoming. Although the aetiology of RA remains unknown a majority of data are consistent with it being mediated via excess pro-inflammatory cytokine production. Given that ES-62 is anti-inflammatory, we hypothesised that it might be able to counter the pathology associated with diseases like RA. Indeed, we found that exposure to ES-62 prevented initiation of collagen-induced arthritis (CIA) in a murine model and also suppressed progression of established disease. Ex vivo analyses demonstrated that these effects correlated with inhibition of TNF-alpha production and inhibition of collagen-specific TH-1 responses. The nematode product was also able to suppress pro-inflammatory cytokine release in vitro in synovial cells derived from RA patients. ES-62 thus constitutes a pathogen-derived immunomodulator with significant therapeutic potential.

The low-affinity receptor for IgG, FcgammaRIIb, negatively regulates B cell antigen receptor (BCR)-mediated proliferative signalling. FcgammaRIIb has been reported to mediate this inhibition by uncoupling the BCR from the RasMAPkinase pathway. We now show that FcgammaRIIb-mediated negative feedback inhibition also correlates with induction of an Erk-associated phosphatase activity that reflects the rapid association of Erk and the MAPkinase phosphatase, Pac-1, and dephosphorylation and inactivation of ErkMAPkinase. This mechanism of abrogating ongoing ErkMAPkinase signalling therefore provides a rationale for rapid immune-complex-mediated feedback inhibition of active antigen-driven B cell responses. In addition, FcgammaRIIb signalling also induces the recruitment and activation of the 3'-inositol phosphatase, PTEN, which by antagonising PI 3kinase activity and inhibiting BCR-coupling to the anti-apoptotic kinase, Akt, provides an additional mechanism for FcgammaRIIb-mediated negative regulation of BCR-coupling to ErkMAPkinase, cell survival and proliferation.

This review examines signal transduction pathways mediating agonist-induced contraction of circular muscle in the body of the esophagus and in the lower esophageal sphincter (LES). In the LES, circular muscle agonists activate a well-defined contractile pathway, involving calcium (Ca(2+))-induced activation of calmodulin and myosin kinase, causing phosphorylation of 20-kDa myosin light chains (MLCs) and contraction. In this pathway, phosphorylation and contraction may be modulated by other factors, resulting, for instance, in inhibition of phosphatase activity, which may potentiate MLC phosphorylation. The agonist-activated contractile pathway of circular muscle from the esophageal body is not as well defined, and it is different from the LES contractile pathway, as it depends on activation of a Ca(2+)-independent protein kinase C (PKC), PKC-epsilon. In this pathway, agonist-induced Ca(2+) influx and/or release activate phospholipases to produce second messengers, such as diacylglycerol and arachidonic acid. The second messengers, however, activate a PKC-epsilon and a contractile pathway, which is Ca(2+) independent. This contractile pathway depends on activation of the mitogen-activated protein (MAP) kinases ERK1 and ERK2 and of p38 MAP kinase. These kinases are, in turn, linked to the small heat-shock protein HSP27, to integrin-linked kinase, and perhaps to other Ca(2+)-independent kinases, such as zipper kinase capable of producing MLC phosphorylation and contraction.

Phosphorylcholine (PC) is increasingly becoming recognised as a carbohydrate-associated component of a wide variety of procaryotic and eucaryotic pathogens. Studies employing nematode PC-containing molecules indicate that it possesses a plethora of immunomodulatory activities. ES-62 is a PC-containing glycoprotein, which is secreted by the rodent filarial nematode Acanthocheilonema viteae and which provides a model system for the dissection of the mechanisms of immune evasion induced by related PC-containing glycoproteins expressed by human filarial nematodes. At concentrations equivalent to those found for PC-containing molecules in the bloodstream of parasitised humans, ES-62 is able to inhibit antigen receptor-stimulated proliferation of B and T lymphocytes in vitro and in vivo. The active component of ES-62 appears to be PC, as PC conjugated to albumin or even PC alone broadly mimic the results obtained with ES-62. PC-induced impaired lymphocyte responsiveness appears to reflect uncoupling of the antigen receptors from key intracellular proliferative signalling events such as the phosphoinositide 3-kinase, protein kinase C and Ras mitogen-activating protein kinase pathways. Although PC-ES-62 can desensitise B and T cells, not all cells are affected, and in fact it is still possible to generate an antibody response to the molecule. Dissection of this response indicates that it is of the TH-2 type. This appears to reflect the ability of ES-62 to direct the polarity of the T cell response by suppressing the production of proinflammatory cytokines, inducing the induction of anti-inflammatory cytokines and by driving the maturation of dendritic cells that direct TH-2 T cell responses.

Keywords: Photomedicine group Reference LPAS-ARTICLE-2001-009 Record created on 2007-07-20, modified on 2016-08-08

Background & Aims: In cat spontaneous lower esophageal sphincter (LES), tone is maintained by the activity of group I secreted phospholipase A2 (sPLA2-I) that produces arachidonic acid. Arachidonic acid metabolites activate G proteins linked to phospholipases, producing second messengers and activation of a protein kinase C–dependent pathway to maintain tone. We examined the role of sPLA2-I in the maintenance of tone in human LES samples obtained from organ donors. Methods: In vitro LES tone and sPLA2-I–induced contraction of enzymatically isolated LES smooth muscle cells were measured in the absence or presence of inhibitors. Cell permeabilization by saponin allowed use of G-protein antibodies. Results: In vitro LES tone was reduced by inhibitors of sPLA2-I, by indomethacin, by the phosphatidylcholine-specific phospholipase C inhibitor D609, and by the protein kinase C inhibitor chelerythrine. sPLA2-I–induced contraction of isolated LES smooth muscle cells was reduced by indomethacin, pertussis toxin, Gi3 antibodies, D609, and by chelerythrine. Conclusions: Human LES tone is maintained by the activity of sPLA2-I that produces arachidonic acid and metabolites and activation of Gi3-linked receptors and of phosphatidylcholine-specific phospholipase C, resulting in production of diacylglycerol, activation of PKC, and maintenance of tone through a protein kinase C–dependent contractile pathway. GASTROENTEROLOGY 2000;119:1243-1252

Phosphorylcholine (PC) is a structural component of a variety of prokaryotic and eukaryotic pathogens. In some cases, PC in infectious agents can benefit the infected host due to its targeting by both the innate and adaptive immune responses. However, as discussed here, PC exhibits a surprising range of immunomodulatory properties that might be to the detriment of the host.

OBJECTIVES Robotic telesurgery has been demonstrated over long distances and offers theoretical benefits to urologic training and the care of patients in remote regions. The multiple arms and three-dimensional vision of the da Vinci robotic system provide a platform conducive to long-distance telementoring and telesurgery. Whereas prior telesurgical efforts have used dedicated lines for information transmission, the public Internet offers a less expensive alternative. It was the intent of this study to test the validity of using the da Vinci system in urologic telesurgery, and to conduct telerobotic nephrectomies using the public Internet. METHODS We performed four right nephrectomies in porcine models using the da Vinci robotic system. Telementoring and telesurgical approaches were used, with resident surgeons operating a console adjacent to the swine, while attending surgeons simultaneously operated a second console at distances of 1300 and 2400 miles from the operating room. RESULTS All four procedures and both telementoring and telesurgical models were successful. Round-trip delays from 450 to 900 ms were demonstrated. Blood loss was minimal, and there were no intraoperative complications. CONCLUSIONS This study represents the first use of the da Vinci Surgical System in urologic telesurgery and the first successful telesurgical nephrectomy in an animal model.

Theileria parva-infected B cells express Jagged-1 and activate Notch signalling in a parasite-dependent manner. ES-62, a filarial nematode-secreted phosphorylcholine-containing glycoprotein, is able to further stimulate Notch-mediated signalling in parasitized cells. Notch is also activated to a similar extent by addition of exogenous IL-10, and this occurs prior to any increase in proliferation in T. parva-infected B cells.

Esophageal reflux is a common condition that affects children and 1 in 10 adults, and if untreated may result in chronic esophagitis, aspiration pneumonia, esophageal strictures, and Barrett's esophagus, a premalignant condition. Although esophagitis is a multifactorial disease that may depend on transient lower esophageal sphincter (LES) relaxation, speed of esophageal clearance, mucosal resistance, and other factors, impairment of LES pressure is a common finding in patients complaining of chronic heartburn. Our data suggest that esophageal and LES circular muscle utilize distinct Ca2+ sources, phospholipid pools, and signal transduction pathways to contract in response to acetylcholine (ACh): (1) In esophageal muscle ACh-induced contraction requires influx of extracellular Ca2+ and may be linked to phosphatidylcholine metabolism, production of diacylglycerol (DAG) and arachidonic acid, and activation of a protein kinase C (PKC)-dependent pathway. (2) In LES muscle ACh-induced contraction utilizes intracellular Ca2+ release arising from metabolism of phosphatidylinositol (PI), and a calmodulin-myosin light chain kinase-dependent pathway. Resting LES tone, on the other hand, may be due to relatively low basal PI hydrolysis resulting in submaximal levels of inositol triphosphate (IP3)-induced calcium release and interaction with DAG to activate PKC. (3) After induction of experimental esophagitis, basal levels of PI hydrolysis and intracellular calcium stores are substantially reduced, resulting in a reduction of resting tone. In addition the signal transduction pathway responsible for LES contraction in response to ACh changes from one that depends on IP3 production, calcium release, and calmodulin activation to one that relies on influx of extracellular calcium and activation of PKC.

Milkfat was extracted quantitatively from dairy foods and the vitamins A, E and beta-carotene in the extract were analysed by normal phase high performance liquid chromatography (HPLC) using a silica column, a fluorescence detector and a UV/visible detector. Products such as butter and anhydrous milkfat were dissolved simply in hexane, whereas milk and milk powders underwent a Rose-Gottlieb fat extraction prior to HPLC analysis. The extraction method used as superior to commonly used saponification procedures in terms of speed as well as reduced loss and isomerization of the vitamin components in the food, because the chemical form of the vitamin is not changed to a less stable form such as retinol during extraction. The individual geometrical isomers of vitamin A, which possess different in vivo vitamin activities, were uniquely identified and quantified. Recovery studies showed satisfactory results for all three vitamins. The simultaneous nature of the assay allowed the estimation of the total vitamin A activity of the food because both vitamin A and pro-vitamin A (beta-carotene) were quantified. The single extraction and single HPLC analysis used for the three vitamins made the method well suited for routine work. (C) 1996 Elsevier Science Limited

The relative roles of G-proteins and protein tyrosine kinases (PTKs) in the regulation of antigen receptor-mediated signalling in B and T cells is controversial. As they, and the biochemical events they control, are potential targets for intervention in various immune dysfunctions, the resolution of the controversy is of great interest. Here, Margaret Harnett and Kevin Rigley provide a timely assessment of current understanding, and propose a model for the interaction of G-proteins and PTKs in antigen receptor-mediated signalling.

this issue, Ref. 1). Ligation of the BCR induces protein tyrosine kinase (PTK) activity I-s, as well zs phospholipase C (PLC)-mediated hydrolysis of phos- phatidylinositol (4,5)-bisphosphate (PIP2)to generate inositol (1,4,5)-trisphosphate (IP 3) and diacylglycerol (DAG) (Refs 1,6). IP 3 mobilizes intracellular stores of calcium ([Ca2+]0, leading to activation of calcium/ calmodulin-dependent protein kinase II (CaM-KII), and DAG is an activator of the ~, 13 and ~ isoforms of protein kinase C (PKC) (~, 13, ~, l~ and "q isoforms are expressed in B cellsV). Although the data are controversial, genetic and biochemical studies I,sq° indicate that the primary signal in this pathway may be the activation of the sIg-associated PTK p72 ~yk, resulting in the binding and activation of a srorelated PTK(s) that, in turn, tyrosine phos- phorylates conserved sequences termed antigen- receptor homology 1 (ARH1) motifs in the cyto- plasmic domains of Ig-a and Ig-13 (Refs 1,11). This induces recruitment of additional p72 ~yk, as well as the reorientation and enhanced binding and acti- vation of the ARHl-associated

Background & Aims Patients with eosinophilic esophagitis (EoE) often become dysphagic from the combination of organ fibrosis and motor abnormalities. We investigated mechanisms of dysphagia, assessing the response of human esophageal fibroblasts (HEFs), human esophageal muscle cells (HEMCs), and esophageal muscle strips to eosinophil-derived products. Methods Biopsy specimens were collected via endoscopy from the upper, middle, and lower thirds of the esophagus of 18 patients with EoE and 21 individuals undergoing endoscopy for other reasons (controls). Primary cultures of esophageal fibroblasts and muscle cells were derived from 12 freshly resected human esophagectomy specimens. Eosinophil distribution was investigated by histologic analyses of full-thickness esophageal tissue. Active secretion of EoE-related mediators was assessed from medium underlying mucosal biopsy cultures. We quantified production of fibronectin and collagen I by HEF and HEMC in response to eosinophil products. We also measured the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 by, and adhesion of human eosinophils to, HEFs and HEMCs. Eosinophil products were tested in an esophageal muscle contraction assay. Results Activated eosinophils were present in all esophageal layers. Significantly higher concentrations of eosinophil-related mediators were secreted spontaneously in mucosal biopsy specimens from patients with EoE than controls. Exposure of HEFs and HEMCs to increasing concentrations of eosinophil products or co-culture with eosinophils caused HEFs and HEMCs to increase secretion of fibronectin and collagen I; this was inhibited by blocking transforming growth factor β1 and p38 mitogen-activated protein kinase signaling. Eosinophil binding to HEFs and HEMCs increased after incubation of mesenchymal cells with eosinophil-derived products, and decreased after blockade of transforming growth factor β1 and p38 mitogen-activated protein kinase blockade. Eosinophil products reduced electrical field–induced contraction of esophageal muscle strips, but not acetylcholine-induced contraction. Conclusions In an analysis of tissues samples from patients with EoE, we linked the presence and activation state of eosinophils in EoE with altered fibrogenesis and motility of esophageal fibroblasts and muscle cells. This process might contribute to the development of dysphagia.

Gastro-esophageal reflux disease (GERD) is a common disorder in the US. Up to 60% of GERD patients have non erosive reflux disease (NERD) i.e. reflux-associated pain with normal-appearing esophageal mucosa. In contrast erosive esophagitis (EE) is characterized by inflammation, i.e. infiltration of leukocytes in the mucosa. We have shown that increased expression of the acid-sensitive receptor TRPV1 occurs in both groups of patients, but only biopsies from EE patients exhibit upregulation of cytokines and mucosal infiltration by leukocytes. The mechanisms responsible for these changes were examined in human esophageal epithelial (HET-1A) cells, and compared to data obtained in patient biopsies. We show that repeated (7 times in 48 h) acid exposure (pH 5.0, 12 min) (EE protocol) results in mRNA upregulation for TRPV1, Lyso-PAF AT, IL-8, eotaxin-1, -2, and -3, macrophage inflammatory protein-1alpha and monocyte chemoattractant protein-1, as it occurs in EE biopsies. In contrast exposure (7 times in 48 h) of esophageal epithelial cells to a less acidic pH (5.8) for shorter time periods (8 min) (NERD protocol) induces mRNA upregulation for TRPV1 only, mimicking changes found in NERD biopsies. We next examined the transcription factors activated by the two HCl exposure protocols. CREB, a cAMP response element-binding protein, is a Ca2+-dependent transcription factor. TRPV1 activation induces Ca2+ influx into neurons and esophageal epithelial cells, leading to activation of Ca2+mediated signal transduction, including activation of Ca2+/calmodulin-dependent protein kinase (CaMK). CaMK activates CREB via phosphorylation of Ser-133, that is required for CREB-activated transcription. We find that CREB phosphorylation and TRPV1 upregulation occurs both in the NERD and EE model of HCl exposure. NF-kB is a family of Rel domaincontaining proteins that is thought to participate in production of cytokines. In the cytoplasm NF-kB is in an inactive state, containing an IkBalpha (inhibitor of kappa Β) protein. IkBalpha inactivates the NF-kB transcription factor by masking the nuclear localization signals of NFkB proteins and keeping them sequestered in an inactive state in the cytoplasm. NF-kB is activated by proteolitic cleavage allowing removal of the phosphorylated IkB and subsequent translocation of Rel dimers to the nucleus. Phosphorylation of IkB occurs through phosphorylation/activation of an IkB kinase IkkB. We find that NF-kB activation, demonstrated by phosphorylation/activation of IkkB occurs in the EE but not in the NERD model of HCl exposure. These data are consistent with CREB-dependent transcription affecting TRPV1 upregulation in the NERD and EE models of HCL exposure, and NF-kB-dependent transcription affecting EE and related cytokine/LysoPAF AT upregulation. β

Long-term effects of tafamidis : a new therapeutic option for patients with transthyretin familial amyloid polyneuropathy (ttr-fap)

G A A b st ra ct s for the study were chest pain (n=6), dysphagia (52), reflux (22), dysphagia and reflux (8) and other (10). There was 100% concordance of the diagnoses of normal (21) and achalasia (18). No case of nutcracker was diagnosed by either classification. A motor abnormality by the conventional classification includes several of the categories described by the Chicago classification. In each case the abnormality was described in detail in the report. Of the 18 achalasia pts (ages 14-87), 5 were type I, 13 were type II and 1 was type III. Of the 5 type I pts, 4 underwent myotomy but 2 continue with symptoms, one has not been treated. Of the 11 Type II pts, 9 are doing well after treatment. The outcome was not different between the types (Fisher's exact test, p=0.27). Conclusions: Application of the Chicago classification to HRM analysis does not change the diagnosis compared to conventional classification. It further subclassifies the motor abnormalities but may not be clear to referring physicians without a descriptive explanation. Though not statistically significant, the treatment outcome in our limited number of patients is supportive of the value of subclassification in achalasia.