Response to the dental scientist model.

In general, considerable strength and much realism exist in the model as presented by Dr. McHugh. Progress in biological science is central to any scheme anticipating change in medicine, including dental and oral medicine.

YMC1, a yeast gene encoding a new putative mitochondrial carrier protein.

We have cloned and sequenced a Saccharomyces cerevisiae gene coding for a protein with significant similarities to the mitochondrial carrier family. The gene we termed YMC1 (yeast mitochondrial carrier) is located on chromosome XVI, closely downstream of ARO7 encoding chorismate mutase.

Kinetic analysis of rat parotid gland muscarinic receptors in vivo: comparison with brain and heart.

(RR)- and (SS)-quinuclidinyl iodobenzilate enantiomers [(RR)- and (SS)-IQNB, active and inert, respectively] have been synthesized for quantitative evaluation of muscarinic acetylcholine receptor (mAChR) binding. Pharmacokinetic approaches have not been used previously to assess in vivo IQNB binding in nonexcitable tissues. We have applied this method to examine mAChRs in rat parotid gland in comparison to those in brain and heart.

Human neoplastic submandibular intercalated duct cells express an acinar phenotype when cultured on a basement membrane matrix.

Culture of the human neoplastic submandibular gland intercalated duct cell line, HSG, on the basement membrane extract Matrigel induces dramatic morphologic changes and cytodifferentiation. Transmission electron microscopy demonstrated an acinar cell phenotype with polarized cells containing a well-developed Golgi apparatus, multiple microvilli-like projections from the apical surfaces into a lumenal-like area, and numerous granule-like organelles. Amylase, an acinar cell marker, was detected by both immunocytochemical and Northern blot analyses.

The functions of saliva.

Oral health is determined to a considerable extent by our ability to produce saliva. Not only must adequate amounts be produced, but a large number of specific proteins also must be secreted for the mouth to function properly. This brief review is directed at describing (1) how saliva is secreted, (2) the consequences of decreased salivary function, (3) the components necessary for oral homeostasis, and (4) the common causes of salivary hypofunction.

Signaling mechanisms that regulate saliva formation.

The precipitating event in the formation of saliva is the binding of neurotransmitter molecules to cell surface receptor proteins. The principal neurotransmitters involved are acetylcholine and norepinephrine that bind, respectively, to muscarinic-cholinergic, and alpha- and beta-adrenergic receptors. The transduction of the extracellular signal requires an integral membrane protein capable of binding GTP, a G protein, that specifically interacts with the receptor.

Mapping the putative RNA helicase genes by sequence overlapping.

An 'electronic' gene mapping procedure based on computer-aided search for overlapping gene sequences was used to identify adjacent genes and localize several putative RNA helicase genes to different chromosomes. PRP28 and AMD1 genes map to the right arm of chromosome IV next to sup2, which encodes a tyrosine tRNA. PRP16, previously mapped to chromosome XI, is tightly linked to MRP-L20.

Elevation of cytosolic [Ca2+] due to intracellular Ca2+ release retards carbachol stimulation of divalent cation entry in rat parotid gland acinar cells.

This study examines the activation of divalent cation entry into rat parotid gland acinar cells by using Mn2+ as a Ca2+ surrogate cation. Following muscarinic-cholinergic stimulation of dispersed parotid acini with carbachol (10 microM), the onset of internal Ca2+ release (cytosolic [Ca2+], [Ca2+]i, increase) and the stimulation of Mn2+ entry (increase in fura2 quenching) are not simultaneously detected. [Ca2+]i elevation, due to intracellular release, is detected almost immediately following carbachol addition and peak [Ca2+]i increase occurs at 6.

Characterization of polyphosphoinositide-specific phospholipase C in rat parotid gland membranes.

Hydrolysis of exogenously added, [3H]inositol-labeled, phosphatidylinositol 4,5-bisphosphate (PIP2) by rat parotid membranes was increased, dose-dependently, by the muscarinic cholinergic agonist carbamylcholine (carbachol) in the presence of guanosine 5'-O-thiotriphosphate (GTP gamma S). The stimulation was inhibited by atropine and guanosine 5'-O-thiodiphosphate (GDP beta S). GTP gamma S alone stimulated PIP2 hydrolysis, with half-maximal activation at 0.

Salivary gland function and glucose metabolic status.

To study the relationship between glucose metabolic status and salivary gland function in different-aged persons, subjects with diabetes mellitus (DM = 11), impaired glucose tolerance (IGT = 26), and controls (n = 26), aged 24 to 93, were examined in the oral physiology component of the Baltimore Longitudinal Study of Aging. All were generally healthy (except DM) and nonmedicated. The controls and subjects with IGT were classified using World Health Organization criteria, and diabetic status was assessed using Hb1Ac levels.

Age-related vulnerability.

This presentation considers the vulnerability of the upper alimentary and respiratory tracts to environmental insults during aging. A specific example, salivary gland secretion, is discussed. Available data suggest that while aging per se does not affect salivary performance adversely, it does appear to compromise the glands in such a way that older persons are more vulnerable to exogenous factors that can reduce secretory capacity and, consequently, diminish oropharyngeal health.

Effects of ionizing radiation and beta-adrenergic stimulation on the expression of early response genes in rat parotid glands.

There is little known about the regulation of gene expression in rat parotid glands after exposure to ionizing radiation. The present studies investigate the effects of in vivo ionizing radiation, with subsequent stimulation of beta-adrenergic receptors by isoproterenol, on parotid gland function and on the expression of the early response genes, c-fos, c-jun, and jun B. Ionizing radiation diminished parotid gland weight and saliva output.

Membrane potential modulates divalent cation entry in rat parotid acini.

This study examines the effect of membrane potential on divalent cation entry in dispersed parotid acini following stimulation by the muscarinic agonist, carbachol, and during refill of the agonist-sensitive internal Ca2+ pool. Depolarizing conditions (addition of gramicidin to cells in Na(+)-containing medium or incubation of cells in medium with elevated [K+]) prevent carbachol-stimulated hyperpolarization of acini and also inhibit carbachol activation of Ca2+ and Mn2+ entry into these cells. Conditions promoting hyperpolarization (cells in medium with Na+ or with N-methyl-D-glucamine instead of Na+) enhance carbachol stimulation of divalent cation entry.

Effect of ionizing radiation on sympathetic nerve function in rat parotid glands.

Ionizing radiation (IR) irreversibly damages salivary glands. The pathologic mechanism is unknown. Previously we reported that parotid serous acinar cells may not be the primary site of damage by IR.

The deduced sequence of the transcription factor TFIIIA from Saccharomyces cerevisiae reveals extensive divergence from Xenopus TFIIIA.

TFIIIA is an RNA polymerase III transcription factor that binds to the internal control region of the 5 S RNA gene as the first step in the assembly of a transcription complex. We have identified the gene encoding TFIIIA from Saccharomyces cerevisiae. Protein synthesized in vitro from the cloned gene has the same size, DNA-binding properties, and transcription factor activity as does purified yeast TFIIIA.

Glandular-like morphogenesis of the human submandibular tumor cell line A253 on basement membrane components.

We have studied the interaction of a human tumor cell line, A253, derived from a submandibular gland carcinoma with a differentiation promoting reconstituted basement membrane extract, Matrigel. When cultured on plastic, these cells maintain a flat, cobblestone, epithelial morphology. On Matrigel, A253 cells initially form a honeycomb network of cords of cells which subsequently thickens.

Further studies of salivary inhibition of HIV-1 infectivity.

An HIV-1/ATH8-cell cytopathic system was used to characterize the previously reported anti-HIV-1 activity of human saliva. Inhibitory activity was demonstrated by monitoring viable cell counts, HIV-1 p24 core antigen, and reverse transcriptase levels. Nonfiltered whole saliva, sterilized by irradiation, protected the ATH8 cells from HIV-1 infection.

Salivary gland function and aging: a model for studying the interaction of aging and systemic disease.

This review describes an approach to examining the interaction of aging and systemic disease on a key aspect of oral physiology, salivation. The approach requires several steps: defining general health, and a specific physiological function, at different ages; defining a disease of interest and the influence of the disease on the specific physiological function; and determining if the disease can affect performance of the physiological function with increased age.

Evidence that M3 muscarinic receptors in rat parotid gland couple to two second messenger systems.

The binding affinities of muscarinic antagonists were compared with their abilities to block carbachol (CCh)-mediated stimulation of Ca2+ mobilization and inhibition of isoproterenol-elicited adenosine 3',5'-cyclic monophosphate (cAMP) accumulation in rat parotid cells. The binding of [3H]quinuclidinyl benzilate (QNB) to membranes was inhibited by antagonists with the following potencies (dissociation constant, nM): atropine (1.1) approximately 4-diphenylacetoxy-N-methylpiperidine methbromide (4-DAMP) (1.

Cellular characteristics of long-term cultured rat parotid acinar cells.

We have successfully maintained and biochemically characterized differentiated rat parotid acinar cells cultured for long periods (6 mo.). The cells were cultured on a reconstituted basement membrane matrix in a medium containing a variety of agents that promote cellular proliferation and differentiation.

Muscarinic receptor antagonist effects in parotid acini and M1-CHO cells: evidence of G protein involvement.

We have examined the effects of the muscarinic antagonist atropine, on non-receptor-mediated activation of intracellular calcium (Ca2+i) mobilization events. In dispersed rat parotid acini, atropine significantly lowered basal inositol trisphosphate levels and delayed AlF4(-)-stimulated Ca2+i elevation in a dose-dependent manner. In a cell line transfected with the m1 muscarinic receptor (M1-CHO), atropine significantly lowered (approximately 60%) AlF4- stimulated Ca2+i elevation.