Evidence for the convergence of β-adrenergic and muscarinic signalling systems at a post-receptor site

AbstractThe β-adrenergic agonist isoproterenol stimulates inositol trisphosphate (IP3) formation and cytosolic Ca2+ ([Ca2+]i) mobilization in rat parotid acini via a cAMP-dependent process. Atropine, a muscarinic antagonist, inhibited these isoproterenol responses without affecting isoproterenol-induced amylase secretion or peak [Ca2+]i and IP3 responses elicited by α1-adrenergic stimulation with epinephrine. Atropine had no effect on isoproterenol-induced [Ca2+]i responses in a cell line which lacked muscarinic receptors and did not alter β-adrenoreceptor ligand binding. These results suggest that the inhibition by atropine results from a post-receptor effect on cAMP-mediated stimulation of phosphatidylinositol 4,5 bisphosphate (PIP2) hydrolysis

Using HSV-Thymidine Kinase for Safety in an Allogeneic Salivary Graft Cell Line

Extreme salivary hypofunction is a result of tissue damage caused by irradiation therapy for cancer in the head and neck region. Unfortunately, there is no currently satisfactory treatment for this condition that affects up to 40,000 people in the United States every year. As a novel approach to managing this problem, we are attempting to develop an orally implantable, fluid-secreting device (an artificial salivary gland). We are using the well-studied HSG salivary cell line as a potential allogeneic graft cell for this device. One drawback of using a cell line is the potential for malignant transformation. If such an untoward response occurred, the device could be removed. However, in the event that any HSG cells escaped, we wished to provide additional patient protection. Accordingly, we have engineered HSG cells with a hybrid adeno-retroviral vector, AdLTR.CMV-tk, to express the herpes simplex virus thymidine kinase (HSV-tk) suicide gene as a novel safety factor. Cells were grown on plastic plates or on poly-L-lactic acid disks and then transduced with different multiplicities of infection (MOIs) of the hybrid vector. Thereafter, various concentrations of ganciclovir (GCV) were added, and cell viability was tested. Transduced HSG cells expressed HSV-tk and were sensitive to GCV treatment. Maximal effects were seen at a MOI of 10 with 50 μM of GCV, achieving 95% cell killing on the poly-L-lactic acid substrate. These results suggest that engineering the expression of a suicide gene in an allogeneic graft cell may provide additional safety for use in an artificial salivary gland device.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/63119/1/10763270152436463.pd

Re-engineering the Functions of a Terminally Differentiated Epithelial Cell in Vivo

Because of their easy access, and important role in oral homeostasis, mammalian salivary glands provide a unique site for addressing key issues and problems in tissue engineering. This manuscript reviews studies by us in three major directions involving re-engineering functions of salivary epithelial cells. Using adenoviral-mediated gene transfer in vivo , we show approaches to i) repair damaged, hypofunctional glands and ii) redesign secretory functions to include endocrine as well as exocrine pathways. The third series of studies show our general approach to develop an artificial salivary gland for clinical situations in which all glandular tissue has been lost.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72101/1/j.1749-6632.1999.tb08512.x.pd

Tissue Compatibility of Two Biodegradable Tubular Scaffolds Implanted Adjacent to Skin or Buccal Mucosa in Mice

Radiation therapy for cancer in the head and neck region leads to a marked loss of salivary gland parenchyma, resulting in a severe reduction of salivary secretions. Currently, there is no satisfactory treatment for these patients. To address this problem, we are using both tissue engineering and gene transfer principles to develop an orally implantable, artificial fluid-secreting device. In the present study, we examined the tissue compatibility of two biodegradable substrata potentially useful in fabricating such a device. We implanted in Balb/c mice tubular scaffolds of poly-L-lactic acid (PLLA), poly-glycolic acid coated with PLLA (PGA/PLLA), or nothing (sham-operated controls) either beneath the skin on the back, a site widely used in earlier toxicity and biocompatibility studies, or adjacent to the buccal mucosa, a site quite different functionally and immunologically. At 1, 3, 7, 14, and 28 days postimplantation, implant sites were examined histologically, and systemic responses were assessed by conventional clinical chemistry and hematology analyses. Inflammatory responses in the connective tissue were similar regardless of site or type of polymer implant used. However, inflammatory reactions were shorter and without epithelioid and giant cells in sham-operated controls. Also, biodegradation proceeded more slowly with the PLLA tubules than with the PGA/PLLA tubules. No significant changes in clinical chemistry and hematology were seen due to the implantation of tubular scaffolds. These results indicate that the tissue responses to PLLA and PGA/PLLA scaffolds are generally similar in areas subjacent to skin in the back and oral cavity. However, these studies also identified several potentially significant concerns that must be addressed prior to initiating any clinical applications of this device.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/63126/1/107632702760240562.pd

Re-engineering Primary Epithelial Cells from Rhesus Monkey Parotid Glands for Use in Developing an Artificial Salivary Gland

There is no satisfactory conventional treatment for patients who experience irreversible salivary gland damage after therapeutic radiation for head and neck cancer or because of Sjögren's syndrome. Additionally, if most parenchyma is lost, these patients also are not candidates for evolving gene transfer strategies. To help such patients, several years ago we began to develop an artificial salivary gland. In the present study, we used a non-human primate tissue source, parotid glands from rhesus monkeys, to obtain potential autologous graft cells for development of a prototype device for in situ testing. Herein, we present 3 major findings. First, we show that primary cultures of rhesus parotid gland (RPG) cells are capable of attaining a polarized orientation, with Na+/K+-adenosine triphosphatase, zonula occludens-1, and claudin-1 distributed in specific domains appropriate for epithelial cells. Second, we show that RPG cells exhibit 2 essential epithelial functions required for graft cells in an artificial salivary gland device (i.e., an effective barrier to paracellular water flow and the generation of a moderate transepithelial electrical resistance). Third, we show that RPG cells can express functional water channels, capable of mediating directional fluid movement, after transduction by adenoviral and adeno-associated virus type 2 vectors. Together these results demonstrate that it is feasible to individually prepare RPG cells for eventual use in a prototype artificial salivary gland.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/63202/1/ten.2006.12.2939.pd

Clouds and the Earth's Radiant Energy System (CERES) algorithm theoretical basis document

The theoretical bases for the Release 1 algorithms that will be used to process satellite data for investigation of the Clouds and the Earth's Radiant Energy System (CERES) are described. The architecture for software implementation of the methodologies is outlined. Volume 4 details the advanced CERES techniques for computing surface and atmospheric radiative fluxes (using the coincident CERES cloud property and top-of-the-atmosphere (TOA) flux products) and for averaging the cloud properties and TOA, atmospheric, and surface radiative fluxes over various temporal and spatial scales. CERES attempts to match the observed TOA fluxes with radiative transfer calculations that use as input the CERES cloud products and NOAA National Meteorological Center analyses of temperature and humidity. Slight adjustments in the cloud products are made to obtain agreement of the calculated and observed TOA fluxes. The computed products include shortwave and longwave fluxes from the surface to the TOA. The CERES instantaneous products are averaged on a 1.25-deg latitude-longitude grid, then interpolated to produce global, synoptic maps to TOA fluxes and cloud properties by using 3-hourly, normalized radiances from geostationary meteorological satellites. Surface and atmospheric fluxes are computed by using these interpolated quantities. Clear-sky and total fluxes and cloud properties are then averaged over various scales

Personal experience of osteoarthritis and pain questionnaires: mapping items to themes

Purpose. The aim of this study was to examine the correspondence between qualitative and quantitative methods of coding experience of pain reported by participants with osteoarthritis (OA) of the knee. Method. A mapping grid was produced to record the correspondence between subthemes that emerged from thematic analysis of interviews with 24 participants with knee OA, and from questionnaire items which were used in a study of 192 knee OA participants. Items were rated according to their degree of correspondence between subthemes and questionnaire items and an overall correspondence score was produced for each subtheme and questionnaire measure. Results. The subthemes that corresponded well with the questionnaire items were those that related to socio-emotional functioning, the overall experience of pain and the impact of pain on physical functioning. The questionnaire items did not relate to participants' knowledge about their condition and their experience of the medical system. Conclusions. The study indicated that many aspects of pain experience reported by patients in qualitative interviews are also assessed by commonly used questionnaire outcome measures for people with pain. However, although participants reported that knowledge about their condition and their experience of the medical system were important aspects of the overall pain experience, these are rarely used as outcome measures. Questionnaires that address these additional aspects ofthe pain experience could be useful to further evaluate the experience of pain and may help to address importance concerns raised by patients with OA of the knee

Salivary PYY: A Putative Bypass to Satiety

Peptide YY3-36 is a satiation hormone released postprandially into the bloodstream from L-endocrine cells in the gut epithelia. In the current report, we demonstrate PYY3-36 is also present in murine as well as in human saliva. In mice, salivary PYY3-36 derives from plasma and is also synthesized in the taste cells in taste buds of the tongue. Moreover, the cognate receptor Y2R is abundantly expressed in the basal layer of the progenitor cells of the tongue epithelia and von Ebner's gland. The acute augmentation of salivary PYY3-36 induced stronger satiation as demonstrated in feeding behavioral studies. The effect is mediated through the activation of the specific Y2 receptor expressed in the lingual epithelial cells. In a long-term study involving diet-induced obese (DIO) mice, a sustained increase in PYY3-36 was achieved using viral vector-mediated gene delivery targeting salivary glands. The chronic increase in salivary PYY3-36 resulted in a significant long-term reduction in food intake (FI) and body weight (BW). Thus this study provides evidence for new functions of the previously characterized gut peptide PYY3-36 suggesting a potential simple and efficient alternative therapeutic approach for the treatment of obesity

Imaging and spectroscopy of arcs around the most luminous X-ray cluster RX J1347.5-1145

The cluster RX J1347.5-1145, the most luminous cluster in the X-ray wavelengths, was imaged with the newly installed Space Telescope Imaging Spectrograph (STIS) on-board HST. Its relatively high redshift (0.451) and luminosity indicate that this is one of the most massive of all known clusters. The STIS images unambiguously show several arcs in the cluster. The largest two arcs (> 5 arcsec in length) are symmetrically situated on opposite sides of the cluster, at a distance of ~ 35 arcsec from the central galaxy. The STIS images also show approximately 100 faint galaxies within the radius of the arcs whose combined luminosity is ~ 4 x 10^11 Lsun. We also present ground-based spectroscopic observations of the northern arc which show one clear emission line at 6730 A, which is consistent with an identification as [OII] 3727 A, implying a redshift of 0.81 for this arc. The southern arc shows a faint continuum but no emission features. The surface mass within the radius of the arcs (240 kpc), as derived from the gravitational lensing, is 6.3 x 10^14 Msun. The resultant mass-to-light ratio of ~1200 is higher than what is seen in many clusters but smaller than the value recently derived for some `dark' X-ray clusters (Hattori et al. 1997). The total surface mass derived from the X-ray flux within the radius of the arcs is ~2.1 - 6.8 x 10^14 Msun, which implies that the ratio of the gravitational to the X-ray mass is ~1 to 3. The surface GAS mass within this radius is ~3.5 x 10^13 Msun, which implies that at least 6% of the total mass within this region is baryonic.Comment: 3 figures. Replaced with the final version as appears in the Astrophysical Journal Letters (Jan 10, 1998 issue). This incorporates some important revision