Degranulation of rat salivary glands following treatment with receptor-selective agonists.

1. The aim of this study was to find a drug that induces an almost complete degranulation of secretory cells in rat parotid and submandibular glands. 2. Phenylephrine (alpha-adrenergic), isoproterenol (beta-adrenergic) and mecholine (muscarinic cholinergic) were tested. Time and degree of maximal depletion of acinar and granular convoluted tubule cells were determined morphologically. 3. Following phenylephrine-injection (5 mg/kg or 10.2 mg/kg, i.p.), no effect on the acinar granulation level was observed in either of the glands, while about 50-60% granular convoluted tubules were degranulated for at least 120-180 min post-injection. 4. With isoproterenol (5, 10, 40, 70 or 100 mg/kg, i.p.), degranulation of 100% of the acinar cells in the parotid and 80% of the acinar cells in the submandibular gland was observed 90 min post-injection. Granular convoluted tubule cells did not respond to this beta-adrenergic drug. 5. Mecholine (3.75 or 7.5 mg/kg, i.p.) induced mainly degranulation of granular convoluted tubule cells (about 50% after 120 min). Numbers of granulated acinar cells decreased only slightly in both glands (about 10%, 90-120 min). 6. From this study it appears that with a relatively low dosage (5 mg/kg, i.p.) of isoproterenol, a high level of degranulation can be induced in acinar cells of rat parotid and submandibular glands without toxic side effects. Concerning granular convoluted tubules, only moderate degranulation was observed with phenylephrine and mecholine, respectively.

A study of multiple sclerosis patients with magnetic resonance spectroscopic imaging.

Eleven patients with clinically definite MS and three healthy controls were investigated by magnetic resonance spectroscopic imaging. The data sets were analysed for all voxels containing white matter only. We classify these voxels in healthy controls as normal white matter (NWM), and in MS patients as normal-appearing white matter unaffected by MS lesions (NAWM) or white matter affected by MS lesions. The spectra belonging to the voxels were analysed for content of cholines, creatines and N-acetylaspartate (NAA), and compared as a group. It was found that lesions differ from white matter in chemical composition and, moreover, that normal-appearing white matter differs from healthy white matter. Specifically, levels of NAA are lower in patients. There seems to be a linear relation between the composition of white matter and the expanded disability status scale value for the patient. The presence of lactate could not be established, and no unambiguous differences were found between patients with relapsing-remitting and relapsing-progressive disease.

The role of secretory granules in radiation-induced dysfunction of rat salivary glands.

To investigate the possible role of secretory granules in radiation-induced salivary gland dysfunction, rats were pretreated with isoproterenol (5 mg/kg intraperitoneally) to degranulate salivary gland acini. At maximal depletion, salivary glands were locally irradiated with a single dose of 15 Gy of X rays. Parotid and submandibular/sublingual saliva samples were collected before and 1-10 days after irradiation. The lag phase, flow rate, concentrations of potassium and sodium, and amylase secretion were determined. Sham-treated, isoproterenol-treated and irradiated animals provided reference data. In the parotid gland, but not in the submandibular gland, protection against radiation-induced changes in flow rate and composition of saliva occurred after pretreatment with isoproterenol. Combining morphological data from a previous study with data from the current study, it is suggested that improvement of parotid gland function is attributed predominantly to a proliferative stimulus on acinar cells by isoproterenol and not to its degranulation effect. After pretreatment with isoproterenol, an earlier expression of radiation-induced acinar cell damage leading to death was observed, followed by a faster tissue recovery. Thus the proliferative stimulus on acinar cells may accelerate the unmasking of latent lethal damage, resulting in the earlier replacement of dead cells by new, functionally intact cells.

The role of secretory granules in the radiosensitivity of rat salivary gland acini--a morphological study.

The aim of this study was to investigate the radiosensitivity of salivary gland tissue pretreated with isoproterenol to establish a status of depletion of secretory granules in acinar cells at the time of irradiation. Nuclear aberrations and cell lysis were taken as parameters for cell death. Local X irradiation with a single dose of 15 Gy induced comparable early morphological changes in the rat parotid and submandibular glands. During the first day after irradiation, the most obvious changes were degranulation of serous cells and induction of nuclear aberrations in both the secretory (serous as well as mucous) and intercalated duct compartment. Subsequently, progressive lysis occurred in secretory units but not in intercalated and striated ducts. Recovery of tissue integrity was observed from day 6. Early radiation-induced cell death was not reduced by isoproterenol-induced degranulation of acinar cells before irradiation. Subsequent recovery from radiation damage seemed to occur earlier in parotid glands but not in submandibular glands pretreated with isoproterenol. From the present study it is concluded that the radiosensitivity of serous salivary gland acini is not dependent on the presence of secretory granules at the time of irradiation. There was some evidence for a faster recovery from radiation damage observed after pretreatment with isoproterenol which may be the result of drug-induced stimulation of cell proliferation.

Radiation-induced cell proliferation in the parotid and submandibular glands of the rat.

Repopulation of tissues with cells at damaged sites is an important feature in the recovery of radiation-induced tissue injury. To obtain insight into the regenerative process in salivary gland tissue, proliferative activity was measured as a function of time in the different epithelial cell compartments of rat parotid and submandibular glands after local X irradiation with a single dose of 15 Gy. Bromodeoxyuridine-labeling indices were determined before and 10 h and 1, 3, 6, 10, 15, 20 and 30 days after irradiation. In both glands, X irradiation caused cell death and cell cycle delay manifested during the first day. Three days after irradiation, cell proliferation started in the intercalated duct. Six days after irradiation, proliferation was also observed in acinar and granular convoluted tubule cells. The striated ducts showed proliferative activity starting at day 6 (parotid) and day 10 (submandibular), respectively. The results of this study suggest that after 15 Gy of X rays repopulation takes place in all cell compartments. From the present study it cannot deduce if these cells are originating solely from progenitor cells residing in the intercalated duct or if cells of the other compartments are also stimulated. Proliferative activity was found to be higher in the intercalated duct compartment of the parotid gland than of the submandibular gland, which may be related to the suggested greater radiosensitivity and thus a greater demand for cell replenishment in the parotid gland.

In vitro NMR micro imaging of the spinal cord of chronic relapsing EAE rats.

Chronic relapsing experimental allergic encephalomyelitis (Cr-EAE) was induced in Lewis rats with an emulsion of guinea pig spinal cord tissue in complete Freund's adjuvant enriched with Mycobacterium tuberculosis H37 RA. The sensitized rats developed Cr-EAE showing two to three relapses during the first 40 days. In vitro transverse T2-weighted spin echo images of the spinal cord of Cr-EAE rats, sacrificed at the clinical height (hind leg paralysis and urinary incontinence) of the third bout and their controls, were compared with the corresponding histopathology. Lesions extended over the entire spinal cord, however, the larger lesions were predominantly present in the cervical and upper thoracic regions. In the white matter only areas of demyelination and large perivascular demyelination were discernable on the MR images. Size and shape of these lesions correlated well with the morphological characteristics revealed by histopathology. Plaques in the ventrolateral funiculus were generally located peripherally, while plaques in the dorsal funiculus were mainly present in the medial part. The NMR images, however, could not distinguish between demyelination, remyelination, inflammation, and oedema. Also lesions in the gray matter could not be distinguished with MR imaging techniques. However, if lesions were localized at the interface of the gray and white matter the boundary between the gray and white matter was less well defined.

[MRI and MRS diagnosis. Application in multiple sclerosis patients and research animals with experimental allergic encephalomyelitis]. / MRI- en MRS-onderzoek. Toepassing bij patiënten met multipele sclerose en proefdieren met experimentele allergische encephalomyelitis.

MR imaging and spectroscopy offer a noninvasive way to observe lesions and biochemical changes, respectively, that may provide new insights into demyelinating diseases such as MS. Although the role and importance of some of the metabolites, such as choline and N-acetyl aspartate in brain function and disease are not fully understood, the specificity of these changes may provide information about the stage and reversibility of the brain lesions. Animal studies are particularly insightful when MR spectroscopic signals are correlated with histologic and biochemical techniques. Clinical application of spectroscopic imaging could arise from the capability to differentiate between early lesions that might respond to therapy and older irreversible lesions. Although more data need to be obtained, MR spectroscopy shows promise for monitoring progression of MS and evaluating therapy.

[Xerostomia-related dental caries]. / Xerostomie-gerelateerde cariës.

The initiation and prevention of xerostomia-related dental caries were studied using an in situ model. The morphology of induced carious lesions was compared with that of natural xerostomia-related dental caries. Both natural and induced xerostomia-related dental caries showed the same patterns of decay. Using the in situ model three preventive procedures were tested: neutral F gel applied every second day or weekly, and a daily rinse with a fluoride mouthwash for a period of six weeks. Application of fluoride gel or the use of the fluoride mouthrinse resulted in significant inhibition of the demineralization process. Of the procedures evaluated fluoride gel applied every second day was the most effective in preventing the onset of xerostomia-related dental caries.

[Dry mouth; possible cause--pharmaceuticals]. / De droge mond. De mogelijk oorzakelijke rol van geneesmiddelen.

Many elderly complain about oral dryness. This complaint can be the result of the use of drugs, because many drugs, such as anti-depressives, anti-psychotics and anti-hypertensives, have an anti-cholinergic or anti-adrenergic action. Knowledge of this potential side-effect of drugs as well as adjustment of schedules of drug usage may result in (partly) relief of dryness-related complaints. In this paper first of all mechanisms that underly negative regulation of salivary secretion are discussed. An overview is given of drugs with an salivary secretion inhibitory action. In addition, suggestions are presented to manage drug-induced hyposalivation.

[Cause, effects and treatment of hyposalivation]. / Oorzaak, gevolg en behandeling van hyposialie.

Adverse effects of drugs, radiotherapeutic treatment of head and neck tumors and Sjögren's syndrome are common causes of hyposalivation. The resulting sequelae include oral dryness, burning mouth, impeded oral functions, nocturnal oral discomfort and an increased incidence of rampant caries and oral infections. Causal therapy is only occasionally applicable. The main causes, consequences and management of hyposalivation are described.

Acute irradiation effects on morphology and function of rat submandibular glands.

In this study the morphologic and functional changes were compared after irradiation (single dose, 15 Gy) of rat submandibular salivary glands. Before and 1-10 days after local irradiation of the gland region, samples of submandibular saliva were collected after stimulation by pilocarpine. At the same time-points and also 3 h postirradiation submandibular glands were carefully extirpated and prepared for histocytologic examination (LM, TEM). Maximal increase of the lag phase and decrease of the flow rate were observed 3 days after irradiation, while [K+] and [Na+] increased and decreased, respectively, from days 1 and 3 after irradiation. Morphologic changes were observed from the third hour after irradiation, were maximal 3 days after irradiation and had partially recovered by day 10. Three hours and 1 day after irradiation degranulation of convoluted granulated tubes (CGT) was observed. Three days after irradiation the most striking morphologic changes in serous and mucous cells were distension of the cisternae of the RER, degeneration of mitochondria and vacuolization of the cytoplasm. Fibril-like condensations of electron dense material in the mucous granules were observed 3 h, 1 and 6 days after irradiation. Regranulation of CGT cells was observed from day 6. From this study it is concluded that changes in salivary gland function can be observed before major morphologic changes occur. Functional changes persist after the morphologic changes seem to have virtually returned to normal.

Effects of split-dose X irradiation on rat salivary gland function.

The effect of a single local dose of 15 Gy on salivary gland function in male Albino Wistar rats was compared with the effect of two doses of 7.5 Gy. The intervals chosen were 0-24 h and 1 week. Before and 1-30 days after the last radiation dose, samples of parotid and submandibular saliva were collected simultaneously after stimulation of the glands with pilocarpine. Irradiation with the single dose resulted in an increased lag phase and potassium concentration, and a decreased flow rate and sodium concentration. The rate of secretion of amylase was decreased during Days 1-6, increased at Day 10, and was decreased again at Day 30. With two dose fractions, substantial dose-sparing effects on lag phase, flow rate, and secretion of amylase were observed for both the very early (0-6 days postirradiation) and later (6-30 days postirradiation) effects. These effects were maximal when the interval between the fractions was 6 h. A significant dose-sparing effect on electrolytes was observed for the later effects only, again with a maximum for the 6-h interval. The dose-sparing observed for the very early effects cannot be explained satisfactorily by repair of sublethal damage (SLD), redistribution of cells over the cell cycle, or repopulation of salivary gland tissue between the doses. In contrast to the earlier dose-sparing effects, the split-dose recovery seen for later damage may be attributed, in part, to SLD repair in providing for greater reproductive survival of intercalated ductal cells and enhanced tissue regeneration.

pH measurements with an ion sensitive field effect transistor in the mouth of patients with xerostomia.

A transistor pH electrode (ion sensitive field effect transistor), placed in the upper dentures of eleven xerostomia patients and five healthy volunteers, was used to register pH changes in five-, six- and seven-day-old dental plaque. A mouth rinse with a 10% sucrose solution caused a pH fall of about three decades. A significant difference in duration of critical plaque pH was observed: in xerostomia patients, a 10% longer period of pH less than 5.7 was registered during 60 min following a sucrose rinse. Normal oral functions were not influenced by the denture with an integrated electrode. This method is usable for plaque pH registration in xerostomia patients.

A functional and chemical study of radiation effects on rat parotid and submandibular/sublingual glands.

The aim of this study was to monitor composition and rate of secretion of rat parotid and submandibular/sublingual saliva following local single doses of X-rays ranging from 5 to 20 Gy. Pilocarpine-stimulated samples of parotid and submandibular/sublingual saliva were simultaneously collected with miniaturized Lashley cups before and 1-30 days after irradiation. The lag phase (period between injection of pilocarpine and start of the secretion) and flow rate were recorded and the concentrations of sodium, potassium, calcium, phosphate, and amylase were measured. With increasing dose and time, the salivary flow rate as well as sodium concentration decreased, while potassium concentrations increased throughout the follow-up period. The lag phase and the concentration of amylase reached their maximum at 3 and 10 days after irradiation, respectively. The changes in lag phase and flow rate were most obvious after doses of 15 or 20 Gy and showed a great similarity for parotid and submandibular/sublingual saliva. No dose-response relationship was observed for the changes in concentrations of calcium and phosphate. It is concluded that for radiation doses of 10 Gy and above, irreversible changes (lag phase, flow rate, potassium, sodium) were observed. A saturation of the irradiation effects (lag phase, flow rate) seems to exist at doses larger than 15 Gy. No significant differences were observed between the radiation-induced functional changes in parotid and submandibular/sublingual salivary gland tissue.

Irradiation-induced changes in secretion and composition of rat saliva.

The aim of this study was to monitor changes in the composition and the rate of secretion of rat whole, parotid and submandibular/sublingual saliva after local irradiation of the salivary gland region with a single dose of 10 Gy. Before and 1-30 days after irradiation, saliva samples were collected after stimulation with pilocarpine. Maximum changes in the latent period, the flow rate and the composition of parotid and submandibular/sublingual saliva were observed three days after a radiation dose of 10 Gy. Partial recovery was seen for the latent period, [Pi], [Ca2+] and concentration of amylase, whereas the flow rate and [Na+] remained low and [K+] remained high. The changes in whole saliva were similar to those observed for parotid and submandibular/sublingual saliva, although some variation was observed due to contamination with other contributions to oral fluid.

Effect of X-ray irradiation on the permeability of bovine dental enamel.

To estimate permeability properties of bovine dental enamel, complex impedance measurements and radioisotope diffusion experiments were carried out before and after X-ray irradiation (single dose, 72 Gy) of enamel specimens. Neither impedance measurements nor diffusion experiments showed significant changes in permeability.

Rheological properties of human saliva.

From measurements with a Couette-type viscometer provided with a guard ring it was shown that at the saliva-air interface a protein layer is adsorbed. Measurements of the surface shear modulus of this layer on saliva of 7 healthy subjects were performed at a frequency of about 70 Hz and a temperature of 25 degrees C. For a surface age of about 1.5 h the surface shear modulus and the surface viscosity were in the order of 1 Nm-1 and 10(-3) Nm-1 s, respectively. From ellipsometric measurements it was found that the thickness of the protein layer was approx. 100 nm and, using this value, it could be concluded that the shear modulus and the dynamic viscosity were in the order of 10(7) Pa and 10(4) Pa s, respectively. The layer appeared to be fragile. Even shear deformation amplitudes of 4 x 10(-5) are too high to assure linearity. The complex viscosity (eta = eta' - i eta") of the bulk liquid of human submandibular saliva below the absorbed layer was measured in the frequency range 70 Hz-200 kHz with 3 torsional resonators, each for a different frequency, and a Ni-tube resonator. It was concluded, that the real part of the complex viscosity (eta') decreases from 1.1 mPa s at 70 Hz to a value of 0.95 mPa s at high frequencies. Except at the lowest frequency (70 Hz), the value of eta" was too small to be detected.