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Secretion of saliva in X-irradiated rat submandibular glands.

The mechanism of radiation-induced dysfunction in rat submandibular glands was investigated at the cellular level. After X irradiation (single dose, 15 Gy), a vacuolation in the acinar cells or an enlargement of the acinar lumen was observed as a typical morphological change for 2 weeks. As observed using a video-enhanced contrast differential interference contrast (VEC-DIC) microscope, exocytosis and shrinkage of the acinar cells induced by application of pilocarpine (100 microM) were markedly suppressed for 5 days and then recovered to 80% of the control levels. Using an immunohistochemical method, no significant change was observed in amylase distribution, but a marked loss of aquaporin 5 was found in the acinar cells after the irradiation. The extent and time course of pilocarpine-induced mobilization of intracellular Ca(2+) did not change after the irradiation. We conclude that radiation-induced dysfunction in the salivary glands is due to an impairment of exocytosis and a reduction of water secretion. The loss of aquaporin 5 and possibly other membrane-fusion proteins in acinar cells may be the major mechanism underlying such a dysfunction.

Amylases↗

The role of cyclic AMP response element-binding protein in testosterone-induced differentiation of granular convoluted tubule cells in the rat submandibular gland.

The postnatal development of granular convoluted tubules (GCT) in the duct system of the rodent submandibular gland is known to be androgen-dependent, but the underlying molecular mechanism is unclear. To test the possible role of the transcription factor, cyclic AMP response element-binding protein (CREB), in the androgen-induced differentiation of GCT, the effect of testosterone on the expression and localization of epidermal growth factor (EGF), a marker of GCT cells, and of CREB was examined in the submandibular glands of immature 3-week-old rats. Northern blotting demonstrated increases in both EGF and CREB mRNA 1-4 days after testosterone administration. Immunoprecipitation also indicated that CREB protein was increased in amount with testosterone administration, and that induced CREB was phosphorylated at the serine residue as in the active form of CREB. In situ hybridization demonstrated that cells with CREB mRNA signal first appeared in the distal portions of striated ducts at 1 day and had increased in number by 4 days after giving testosterone, when cells with EGF mRNA signal became evident in the same duct portions. Immunohistochemistry also showed the occurrence of CREB protein in the nuclei of duct epithelial cells before their differentiation into EGF-positive GCT cells. Finally, pieces of submandibular gland from immature rats were cultured in vitro and their expression of EGF mRNA analysed by the reverse transcriptase-polymerase chain reaction. Testosterone in the medium caused a marked enhancement of EGF expression in the gland in 1-4 days, which was attenuated by simultaneous administration of the antisense oligonucleotide for CREB as well as that for the androgen receptor. These results suggest the CREB is upregulated by androgen and has a crucial role in androgen-induced differentiation of GCT in the duct system of the rat submandibular gland.

Androgen Receptor Antagonists↗

Aryl hydrocarbon hydroxylase activity in rat submandibular glands.

The activities of aryl hydrocarbon hydroxylase and 7-ethoxycoumarin O-de-ethylase in the rat submandibular gland, both of which are normally very low, were increased about 10- and 25-fold, respectively, by administration of 3-methylcholanthrene. The increased activity of aryl hydrocarbon hydroxylase was inhibited by 7,8-benzoflavone but little if any by metyrapone and SKF-525A. The submandibular gland aryl hydrocarbon hydroxylase was also inhibited by antibodies elicited against rat liver cytochrome P-448 and NADPH-cytochrome P-450 reductase, respectively, and slightly by anti-cytochrome b 5 antibody, suggesting that submandibular gland aryl hydrocarbon hydroxylase resembles that of the liver, not only in the immunochemical properties of cytochrome P-450 involved, but also in the entire enzyme complex, including the electron transferring system. Cofactor(s) required by aryl hydrocarbon hydroxylase in submandibular gland was NADPH, whereas NADH sustained the reaction at a much slower rate (about 20%). NADH showed some additive action when added with NADPH.

7-Alkoxycoumarin O-Dealkylase↗

Giant mitochondria in a pleomorphic adenoma of the submandibular gland.

A benign pleomorphic adenoma of the submandibular gland was examined by electron microscopy. In some areas, the epithelial cells comprising the tumor formed ductlike structures surrounding a lumen filled with membrane vesicles. The cells actually abutting the lumens had giant mitochondria measuring up to 8 micrometers in diameter; such enlarged organelles were absent from immediately subjacent cells. The giant mitochondria exhibited a variety of cristal arrangements, the most common being a quasireticulate one. They often contained expanded cristae that enclosed a number of helical filaments. Bundles of 14-nm tubules with faintly discernible axial periodicity were frequently present in the matrix compartment, as were amorphous dense inclusions. The basis for the occurrence of giant mitochondria only in duct cells may reside in microenvironmental factors rather than in altered nuclear or mitochondrial genomes.

Adenoma, Pleomorphic↗

Mitotic proliferation of myoepithelial cells during regeneration of atrophied rat submandibular glands after duct ligation.

BACKGROUND: The purpose of the present study was to elucidate whether myoepithelial cells proliferate mitotically during regeneration of rat submandibular glands after atrophy. METHODS: The excretory duct of the right submandibular gland of rats was doubly ligated near the hilum with metal clips, which were removed after 7 days of ligation (day 0). The regenerating right submandibular glands were removed from 0 to 14 days after removal of the clips. The removed tissue was examined with immunohistochemical double staining for proliferating cell nuclear antigen (PCNA) as a marker of proliferating cells and actin as a marker of myoepithelial cells, as well as with transmission electron microscopy (TEM). RESULTS: The PCNA-positive myoepithelial cells were observed at the periphery of transitional duct-acinar structures, ducts and acini in the regenerating glands at every time-point, and the PCNA-labeling index of myoepithelial cells increased greatly especially between day 2 and 4. The mitosis of myoepithelial cell was also identified by TEM at day 4. CONCLUSION: These findings suggest that myoepithelial cells are able to proliferate mitotically during regeneration of rat submandibular gland.

Actins↗

Submandibular gland duct endoscopy. Diagnostic value for salivary duct disorders in comparison to conventional radiography, sialography, and ultrasonography.

The purpose of this study is to evaluate the usefulness of endoscopy as a procedure for the diagnosis of submandibular gland duct disorders. Endoscopy of the submandibular glands was performed on 12 patients with symptoms of obstructive sialoadenitis to identify the cause of obstruction. The endoscopic findings were then compared to those of diagnostic procedures such as conventional radiography, sialography, and ultrasonography. Six normal subjects also underwent endoscopy to better understand the normal findings of the duct system. Endoscopy demonstrated salivary gland calculus in 5 of 12 patients, which was revealed as filling defects on sialograms and as strongly echogenic structures on ultrasonograms in 4 of the patients. Endoscopy revealed secretion plugs, secretion plaques, and/or stenosis, which could not be seen by any other diagnostic procedures in 5 patients, as the cause of recurrent swelling in all 7 patients not demonstrating sialolith. Abnormal findings of the duct wall such as vasodilatation, fibrosis, edema, or erythema were seen in four patients, three of whom exhibited dilatation of the duct system on sialograms. In four patients, a decreasing internal echo level of the gland was seen on ultrasonograms. Our initial results for submandibular gland duct endoscopy thus appear to be promising.

Adult↗

Chronic aspiration in children. When are bilateral submandibular gland excision and parotid duct ligation indicated?

OBJECTIVE: To assess the efficacy of bilateral submandibular gland excision and bilateral parotid duct ligation in treatment of chronic aspiration in neurologically impaired children. DESIGN: Retrospective chart review and telephone follow-up interview. SETTING: Tertiary care center. PATIENTS: All patients from 1986 through 1994 who underwent bilateral submandibular gland excision and bilateral parotid duct ligation and had evidence of at least 1 episode of aspiration pneumonia in the year prior to undergoing surgery. MAIN OUTCOME MEASURE: Two main outcomes measures were (1) the change in number of hospitalizations for pneumonia and total number of lower respiratory tract infections between 1 year before and 1 year after surgical intervention and (2) telephone assessment of patient outcome with respect to parental satisfaction, effect on quality of life, care requirements, amount of suctioning, and use of voice. RESULTS: Sixteen patients aged 16 months to 18 years were included. After surgical intervention, there was a significant decrease in the mean (+/-SD) number of pneumonias (2.3 +/- 1.44 before surgery, 0.9 +/- 1.2 after surgery; P < .001) and hospitalizations (1.2 +/- 0.8 before surgery, 0.4 +/- 0.8 after surgery; P < .005). Six patients had a tracheostomy at the time of surgery, and 1 required a tracheostomy 2 years after surgery. No individual required laryngotracheal separation. Eleven families were able to be contacted by telephone. Caretakers reported that in 8 of 11 patients, quality of life was improved and care requirements decreased. Seven patients used voice for at least some degree of communication. Three patients had postoperative complications involving the parotid glands; all resolved after further therapy. CONCLUSION: Bilateral submandibular gland excision and bilateral parotid duct ligation reduce the incidence of aspiration pneumonias and hospitalization, and decrease overall care requirements in a select group of neurologically impaired children. Because they are voice sparing, are efficacious, and have a low morbidity, they should be considered before laryngotracheal separation or tracheoesophageal diversion.

Adolescent↗

Immunohistochemical localization of carbonic anhydrase isoenzymes VI, II, and I in human parotid and submandibular glands.

Human salivary carbonic anhydrase (HCA VI) was purified by inhibitor affinity chromatography and its location in the human parotid and submandibular glands identified, using a polyclonal antiserum raised against the purified enzyme in rabbits in conjunction with the peroxidase-antiperoxidase complex method. The antibodies raised against the purified enzyme in rabbits did not crossreact with the HCA II or I. However, they slightly recognized human IgA; the antiserum was therefore absorbed with human IgA before immunohistochemical use. HCA VI-specific staining was detected in the cytoplasm and particularly in the secretory granules of the serous acinar cells of both parotid and submandibular glands, the staining of the secretory granules being most distinct in paraformaldehyde-fixed tissues. Some epithelial cells and the luminal content of the striated ducts also gave a specific HCA VI staining. Staining specific for HCA II was also found in the granules of the serous acinar cells, particularly in the submandibular gland when Carnoy fluid fixation was used. Slight HCA II-specific staining was also detected in the striated ductal cells in the Carnoy fluid-fixed specimens. No staining specific for HCA I was detected. The results indicate that the serous acinar cells in human parotid and submandibular glands contain abundant HCA II and HCA VI. Interestingly, only HCA VI is secreted into the saliva, although both enzymes appear to be located in structures resembling the secretory granules in the acinar cells. The enzymes probably form a mutually complementary system regulating the salivary buffer capacity.

Antibodies↗

Prevention of liquid-diet-induced damages on submandibular glands by selenium supplementation in rats.

This study was designed to explore the protective effects of selenium on submandibular glands of rats that were maintained on liquid diet. The animals kept on liquid diet for 10 days following 20 days of solid diet, had severe degeneration on both acinar and ductal cells: pycnotic nuclei, enlarged granular endoplasmic reticulum, swollen mitochondria with separated cristae, reduction of secretory granules and abundant lipid droplets were observed. Body and gland weights of these animals were found to decrease significantly compared to those of control group given solid diet for 30 days. The animals that were fed 20 days with solid diet + 10 days liquid diet and also given selenium 0.4 mg/kg for 30 days, did not have any remarkable sign of degeneration on submandibular glands, and their gland weights significantly increased when compared to the animals fed with liquid diet without selenium supplementation. Submandibular glands of the group that was maintained on solid diet for 30 days and supplemented with selenium in drinking water were found to be normal. They showed consistent gain in both body and gland weights almost equal to that of control group. As a result, we assume that adding selenium in liquid diet can prevent the detrimental effects of liquid diet consumption on submandibular glands.

Animal Feed↗

Immunochemical and in situ hybridization analyses of retinoic acid receptor alpha, beta, and gamma in murine Harderian and submandibular glands.

Retinoic acid (RA), through its cognate receptors (retinoic acid receptors, RARs), plays an important role in the ontogenesis and maintenance of the normal function of murine Harderian and submandibular glands. In the present study, autoradiography was used to study RA binding to these glands. Both glands showed high radioactive labelling after [14C]-RA administration in normal and partially vitamin A-deficient (VAD) mice. The peak uptake was at 6 h after [14C]-RA administration in normal mice and at 0.5 h in VAD mice. At 24 h, RA binding remained high in normal mice, while it decreased significantly in VAD mice. In western blots with an antibody recognizing all forms of RARs, a band of molecular weight 51 kDa was seen in homogenates of both glands. Immunohistochemically, RAR staining was found in the nuclei of the glandular cells. The Harderian gland exhibited more intense staining than the submandibular gland. In the latter, the most intense staining was seen in the acinar cells, followed by the intercalated duct cells. The granular convoluted tubule showed weak immunostaining and the striated duct was negative. In the Harderian gland, RAR immunostaining was observed in both type I and II cells, but only part of them stained with RAR antibody. The expression of RAR alpha, beta, and gamma transcripts was studied by in situ hybridization using specific oligonucleotide probes. The cell-specific expression of RAR alpha mRNA in the submandibular gland corresponded to the RAR proteins detected by immunohistochemistry, while the RAR beta transcript was mainly seen in the striated duct. The transcripts of RAR alpha and beta were evenly distributed in type I and II glandular cells of the Harderian gland. RAR gamma labelling was below detectable levels in both glands. This result suggests that RA and RARs regulate the functions of Harderian and submandibular glands in a cell-specific manner.

Animals↗

Submandibular gland surgery: an audit of clinical findings, pathology and postoperative morbidity.

A series of 86 patients who underwent submandibular gland surgery were reviewed retrospectively. Of the 92 glands excised, non-neoplastic disease accounted for 96% of cases. Preoperative clinical findings were inconsistent such that early surgery should be considered for an enlarged non-tender submandibular gland. The incidence of temporary paresis of the lower branches of the facial nerve was 36% with full recovery, on average, 4 months after surgery. The low 'non-identification' approach to the submandibular gland appears to be the technique which offers the least likelihood of permanent damage to the lower branches of the facial nerve.

Adolescent↗

Altered secretion and processing of epidermal growth factor in adrenergic-induced growth of the rat submandibular gland.

The granular convoluted tubule (GCT) cells of the submandibular glands represent a major production site for epidermal growth factor (EGF). This study investigates EGF production in the submandibular glands in relation to beta-adrenergic stimulation. Rats were treated with isoproterenol (beta-agonist), which caused up to a 400% increase in submandibular tissue weight after 3 weeks. The weight increase coincided with marked morphologic changes, with degranulation and an apparent decrement in the number of the GCT cells. Immunostaining against EGF revealed a reduction in the number of EGF-immunoreactive cells. Concomitantly, the glandular contents of 6-kDa EGF decreased from 12.86+/-3.42 nmol/gland (mean+/-S.E.M.) in controls to 0.26+/-0.03 nmol/gland. EGF mRNA levels, expressed relative to total RNA levels, only tended to be reduced after 3 weeks as judged from RT-PCR and in situ hybridization (ISH). The isoproterenol-treated rats had increased output of EGF in the saliva, but the salivary secretion of protein was also increased. In both glandular tissue and saliva, gel filtration revealed partially processed high molecular weight forms of EGF in the isoproterenol-treated rats. These data indicate that isoproterenol treatment leads to a hyperstimulatory state of the GCT cells, which then causes depletion of the cellular stores of mature EGF, and most likely due to a shortened posttranslational transit, incomplete peptide processing.

Adrenergic beta-Agonists↗

Effects of the chorda tympani damage on submandibular glands: scintigraphic changes.

The aim of this study is to compare the scintigraphic results of a denervated submandibular gland with the contralateral normal side in patients with unilateral chorda tympani damage. Sixteen patients (11 women and five men with a mean age of 27) with unilateral proven chorda tympani damage during their previous ear surgery were included in the study. The perfusion ratio (PR), concentration ratio (CR) and stimulated excretion ratio (SER) were calculated scintigraphically and the results from the salivary glands on opposite sides were compared. For submandibular glands, the perfusion ratio (PR), concentration ratio (CR) and stimulated excretion ratio (SER) were found to be 0.65 +/- 0.21, 0.70 +/- 0.21, 0.79 +/- 0.37, respectively. All ratios resulted from statistically decreased radioactivity accumulation on the affected side (p<0.05). Chorda tympani damage negatively affects the function of the ipsilateral submandibular glands despite the absence of atrophy. Dynamic salivary gland scintigraphy is a practical and valuable method of disclosing the decreased capacity of perfusion, concentration and secretion function in unilateral neurological deprivation.

Adolescent↗

[Oral transmuscular adenoma removal of the submandibular gland].

The technique described for the removal of an adenoma in the submandibular gland through an oral-transmuscular approach is based upon the description of Wassmund and Trauner. Results of the authors with eight operations between 1992 and 1993 and the experiences of the Nordwestdeutsche Kieferklinik of Eppendorf University in Hamburg show that the technique used is able to prevent significant scarring and injuries to the lingual and facial nerves. The technique can be used to remove benign tumors of the submandibular gland if they are not too large. Contraindications include microgenia, trismus, extensive inflammation and malignant tumors.

Adenoma, Pleomorphic↗

Transplantation of vascularized submandibular gland in dogs.

PURPOSE: Presently, treatments for xerostomia only target symptoms, as an active therapy method has not been established. Herein, we discuss the possibility of using a submandibular gland allograft technique for the disease. MATERIALS AND METHODS: Using a vascularized submandibular gland transplantation method, we extracted portions of the submandibular gland, including the duct and chorda tympani branches, from beagle dogs and placed them into the submental region of age- and weight-matched dogs. We then measured the amount of saliva secretion and examined the grafted glands histologically. RESULTS: Sufficient quantities of saliva were secreted from the grafted glands with pilocarpine treatment. Histologic findings showed that the acinar cells in the grafted and untreated contralateral glands had some atrophy, as compared with the normal glands; however, periodic acid Schiff staining showed that they produced saliva. CONCLUSIONS: Transplantation of vascularized submandibular glands into dogs was successful and may become a novel treatment strategy for patients with xerostomia.

Animals↗

[Long-term follow-up after autologous submandibular gland transplantation in scarring keratoconjunctivitis with absolute dry eyes].

INTRODUCTION: An autologous submandibular gland can be transplanted to the temporal fossa and following microvascular anastomosis to the temporal artery and vein and implantation of the secretory duct into the superotemporal conjunctival fornix to provide continuous substitute lubrication. Here we report our experiences with this procedure. PATIENTS AND METHODS: In a prospective, controlled clinical cohort study, we investigated the development of the Schirmer- test, fluorescein-break up time (F-BUT), degree of discomfort, use of pharmaceutical tear substitutes, visual acuity, Rose Bengal staining and conjunctival hyperemia in 14 eyes with a successful submandibular gland transplant and 11 dry eyes without salivary lubrication. RESULTS: Over a mean postoperative period of 3.3 years the transplantation group showed significant improvements of the Schirmer-Test, FBUT, use of pharmaceutical tear substitutes, discomfort and Rose Bengal staining up to the last follow-up when compared to the preoperative and control groups. CONCLUSION: Transplantation of an autologous submandibular gland can improve the lubrication and discomfort of absolute tear deficiency on a long term basis, but the salivary tear film only partially stabilises the ocular surface. The procedure is however able to provide symptomatic relief in extreme cases of absolute dry eyes.

Adolescent↗

Effect of thyroid hormone and sex status on epidermal growth factor concentrations in the submandibular gland of a congenitally hypothyroid mouse model.

Studies were performed to explore the role of thyroid hormone and sex status on epidermal growth factor concentrations in the submandibular gland of a congenitally hypothyroid mouse model designated hyt/hyt. The animals were studied at 20, 30 and 40 days of postnatal age. The euthyroid animals were homozygous or heterozygous for the hypothyroid gene. The homozygous euthyroid animals displayed a pattern of submandibular gland epidermal growth factor concentrations similar to those previously described in other mouse species and showed the expected sex differences. The hypothyroid animals had measurable but very low submandibular gland epidermal growth factor concentrations without sexual dimorphism. Serum thyroxine concentrations in the heterozygotes were comparable to those in the homozygous euthyroid animals, yet the animals had a delayed increase in epidermal growth factor concentrations combined with a later expression of female-male differences. The timing of the sex differences in submandibular gland epidermal growth factor concentrations followed a pattern similar to that seen for the timing of the first litter in these three genetically distinct groups. This infers the timing of the onset of puberty and suggests a role of androgens in the changes seen in epidermal growth factor concentrations. We conclude that thyroid hormone and sex status in this mouse model influence the pattern and concentrations of submandibular gland epidermal growth factor concentrations.

Animals↗

Excision of the submandibular gland by an intraoral approach.

To improve the outcome in patients with benign diseases of the submandibular gland, we have developed an entirely intraoral technique for excision of the submandibular gland. This procedure is anatomically safe and can be performed with minimal morbidity. We believe the essential surgical steps are as follows: (1) infiltration with Xylocaine plus epinephrine with an adequate waiting period for hemostasis; (2) careful identification of the submandibular duct/lingual nerve relationship; (3) anterior retraction of the mylohyoid muscle to expose the superficial lobe; (4) superiorly directed, extraoral, manipulation of the submandibular gland; and (5) close and blunt dissection to the gland laterally to avoid injury to the facial artery and vein.

Adult↗