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[Establishment of rat submandibular gland squamous cell carcinoma induced by DMBA].

OBJECTIVE: To study carcinogenesis and development of salivary gland tumor and establish an animal model of submandibular gland (SMG) tumor. METHODS: Histopathological study during carcinogenesis in rat SMGs using (9,10-dimethyl-1,2-benzanthracene (DMBA) was evaluated. A total of 50 male and female Sprague-Dawley (SD) rats of 8 weeks old and 180-200 g weight were obtained from the Animal Center of Henan Medical University. Under pentobarbital sodium anesthesia, the left SMGs were exposed by surgical procedure. A sponge pellet (1.0 mm x 1.0 mm x 1.0 mm, made by authors) was used as the carrier of the carcinogen. The sponge containing 2% DMBA (Fluka, Switzerland)/acetone solution was implanted into the glandular tissue of the left SMGs. Four rats (2 males and 2 females) were killed after every 2 weeks of the DMBA/sponge implantation. The same method of sponge implantation without DMBA was used at the right side of SMG as a control. All rats left were killed after 20 weeks. The SMGs were fixed in 10% formalin buffer solution for 24 hours, and embedded in paraffin, then 4 microns-thick sections were made for histopathological study. RESULTS: The earliest tumor occurred after 4 weeks of implantation of sponge, a total of 21 lateral tumors were induced (10 females, 11 males). There was no tumor found in the controlled SMG. The peak time of tumor genesis was after 8-12 weeks of implantation, all tumors induced were squamous cell carcinomas(SCCs). The induced tumors grew slowly below the mandibles of rats, which were present as nodular masses without capsules, and the borders were not clear. They were slightly hard when palpated. The process of carcinogenesis can be described as following: squamous metaplasia of cyst-like structures occurred, then SCCs were induced and invaded surrounding tissues. No metastasis was observed in regional lymph nodes and other organs. CONCLUSION: SCCs of SMG can be induced by implantation of DMBA. The present study supports the conclusion that all duct segments undergo squamous metaplasia, and therefore may participate in the genesis of neoplasia during experimental carcinogenesis.

9,10-Dimethyl-1,2-benzanthracene↗

Restoration of morphine-induced alterations in rat submandibular gland function by N-methyl-D-aspartate agonist.

The effects of morphine, 1-aminocyclobutane-cis-1,3-dicarboxylic (ACBD; NMDA agonist) and 3-((R)2-carboxypiperazin-4-yl)-propyl-l-phosphoric acid (CPP; NMDA antagonist) and their concurrent therapy on rat submandibular secretory function were studied. Pure submandibular saliva was collected intraorally by micro polyethylene cannula from anaesthetized rats using pilocarpine as secretagogue. Intraperitoneal injection of morphine (6 mg/kg) induced significant inhibition of salivary flow rate, total protein, calcium, and TGF-beta1 concentrations. Administration of ACBD (10 mg/kg) and CPP (10 mg/kg) alone did not influence secretion of submandibular glands. In combination therapy, coadministration of CPP with morphine did not influence morphine-induced changes in salivary function while ABCD could restore all morphine-induced changes. In combination treatment, ACBD prevented morphine-induced reduction of flow rate, total protein, calcium, and TGF-beta1 and reached control levels. It is concluded that morphine-induced alterations in submandibular gland function are mediated through NMDA receptors.

Analgesics, Opioid↗

Long-term outcomes of submandibular gland transfer for prevention of postradiation xerostomia.

BACKGROUND: Xerostomia is a permanent and devastating sequela of head and neck irradiation, and its numerous consequences affect most aspects of the patient's life. A new method of preserving and protecting a single submandibular gland from radiation damage through the Seikaly-Jha procedure (SJP) has recently been described. OBJECTIVE: To report the long-term outcomes of the SJP. DESIGN: Inception cohort. Patients The trial was conducted between February 1, 1999, and February 1, 2002. All patients were followed up through the head and neck cancer clinic at the Cross Cancer Institute. All data were collected by a dedicated research nurse. Salivary function was evaluated at regular intervals with salivary flow studies and questionnaires. RESULTS: Ninety-six patients were enrolled in the study, and 38 had a minimum of 2 years' follow-up. The cohort of 38 patients was composed of 2 groups: 26 patients had preservation of one submandibular gland through the SJP, while the remaining 12 did not. Salivary flow was preserved in the SJP group, in which 83% of patients reported normal amounts of saliva 2 years after radiotherapy, compared with none in the SJP group. There were no disease recurrences on the side of the transferred gland or in the submental space. There were no surgical complications attributed to the transfer procedure. CONCLUSIONS: The SJP prevented xerostomia in 83% of the study patients. The approach appears to be oncologically sound and safe.

Alberta↗

Küttner's tumor of the submandibular glands: report of five cases with fine-needle aspiration cytology.

Küttner's tumor (KT) is a benign tumor-like lesion of the salivary gland that mimics neoplasm clinically because of presentation as a hard mass. Recently, the histomorphological and immunohistochemical findings of this lesion have been analyzed, and differential diagnostic problems relating to salivary gland lymphoma have been discussed. However, currently there is little information on the cytological findings of those lesions. We present cytological findings from five such cases using fine-needle aspiration cytology (FNAC). FNAC of this lesion may present a diagnostic challenge to the cytologist as lesions share some cytologic features with inflammatory process containing numerous lymphoid cells. Smears obtained from two cases contained moderate to large numbers of lymphoid cells without definite cytological atypia, scattered ductal structures, and acinar cell clusters. The remaining three cases showed low cellularity probably attributable to fibrosis that made it difficult to aspirate the cellular element. FNAC findings of scattered ductal structures surrounded by collagens and infiltrated by a mixed population of lymphoid cells, not specific for KT, are highly suggestive of the diagnosis with the appropriate clinical findings. However, a portion of cytological specimens of KT containing relatively large numbers of lymphoid cells should be differentiated from malignant lymphoma arising from the submandibular gland.

Aged↗

[Examination technique in ultrasound calculus detection in the submandibular gland].

Techniques such as orthopantomography and ultrasound are usually employed for examination for submandibular calculi, but we have found transverse scanning of the submandibular gland by ultrasound to be safe and rapid. As the mandible and hyoid bone present a shadow there is a margin for possible error, but this technique enables these areas to be seen and thus checked: a submandibular calculus may be seen as a third shadow between the other two. Comparison showed ultrasound to be superior to orthopantomography. If the ultrasound is negative and there is a strong clinical suspicion of a submandibular calculus orthopantomography is the next step to be employed.

Diagnosis, Differential↗

[Surgical pathology of submandibular gland. Experience in La Rioja community].

The AA. had work through medical histories of 44 patients operated for inflammatory and tumoral pathology of submandibular gland. Between inflammatory diseases they emphasize sialolithiasis (45%), acute submaxillaritis (18%) and Kuttner's tumor (11%). Pleomorphic adenoma accounted for the 50% of tumors. Diagnosis was based on each peculiar history sheet, on imaging procedures (simple X-ray plate, echography, sialography, TC) and extemporaneous biopsy. In all cases a complete exeresis of the gland was performed. In malignoma cases the procedure was completed with a prophylactic neck dissection. Incidence of lasting paralysis of the ramus marginalis mandibulae accounted for 6.8%.

Adolescent↗

Delayed inhibition of gap-junctional intercellular communication in the acinar cells of rat submandibular glands induced by parasympathectomy and cholinergic agonists.

In this study, the effects of parasympathectomy and cholinergic agonists on gap-junctional intercellular communication and salivary secretion were investigated to clarify the involvement of salivary secretion in delayed uncoupling between acinar cells of rat submandibular glands. Gap-junctional intercellular communication was monitored as dye-coupling in the acinar cells of isolated acini by the transfer of Lucifer Yellow CH. Parasympathectomy induced dye-uncoupling in the acinar cells isolated from denervated salivary glands 12 hr after parasympathectomy-induced salivary secretion. Intraperitoneal application of carbachol (CCh), acetylcholine, pilocarpine, but not isoproterenol, stimulated salivary secretion, and then induced dye-uncoupling in the acinar cells 12 hr later. Atropine suppressed both the salivary secretion and delayed dye-uncoupling induced by parasympathectomy and CCh, when atropine was applied intraperitoneally before the induction of salivary secretion. However, atropine did not suppress the delayed dye-uncoupling by intraperitoneal application of CCh, when atropine was injected after the cessation of CCh-induced secretion. These results suggest that delayed inhibition of gap-junctional intercellular communication by parasympathectomy and cholinergic agonists in rat submandibular glands might be related to the change of secretory function after salivary secretion.

Animals↗

Role of laminin-1 and TGF-beta 3 in acinar differentiation of a human submandibular gland cell line (HSG).

Previous studies show that culturing an immortalized human submandibular gland cell line (HSG) on Matrigel, a basement membrane extract, induces cytodifferentiation. We have further defined this model system and identified factors involved in HSG cell acinar development and cyto-differentiation. Acinar development is marked by cell migration into multi-cellular spherical structures, cell proliferation and apoptosis of the centrally localized cells. In addition, functional differentiation was determined by indirect immunofluorescence and immunoblot analysis for cystatin, a salivary gland acinar cell-specific protein found to be produced by differentiated HSG cells. Matrigel contains multiple extracellular matrix proteins, however, laminin-1 was identified as the major matrix component that induced HSG cell acinar development and cytodifferentiation. Antibodies against specific components of Matrigel and against cell surface adhesion molecules were added to cells in culture to identify components important for HSG cell acinar differentiation. Immunostaining of HSG cell acini identified TGF-beta 2 and beta 3 as the predominant isoforms within the cells. Neutralizing antibodies directed against TGF-beta 3 significantly decreased (P < or = 0.0002) the size of acini formed. These results indicate that multiple components, including laminin-1 and TGF-beta 3, contribute to HSG cell acinar development. This model system will be useful to study acinar differentiation and salivary gland-specific protein expression in vitro.

Apoptosis↗

Ultrastructural studies on the bleb formation in seal and rat submandibular gland striated ducts.

For the present study Seal (Phoca vitulina) and rat submandibular gland striated ducts were investigated by electron microscopy. Both normal and stimulated animals (starved for 24 h and then fed 2 h before the tissues were removed) were examined. Basal invaginations of the cell membrane with areas heavily loaded with mitochondria were typical features of both animals. Secretory granules were especially numerous in the apical part of stimulated duct cells. The granules were separated from the luminal membrane of the cells by a condensed area called the spearating zone. Apical protrusions or blebs, which were frequently occurring in striated ducts of both animals, are interpreted as manifestations of apocrine secretion. It was concluded that this way of apocrine secretion is a fundamental function, since seals, which have salivary glands of a rather simple composition with acini which are functionally reduced, have retained the ability to form blebs.

Animals↗

Alterations in nitric oxide synthase activity and expression in submandibular glands of NOD mice.

The non-obese diabetic (NOD) mouse model of autoimmune sialadenitis offers the possibility of studying the L-arginine/nitric oxide signaling pathway in salivary glands in basal and neurotransmitter-stimulated conditions and, thus, of analyzing the neural control of the secretory process in the target organ. The purpose of this study was to explore putative alterations in the activity and expression of nitric oxide synthase (NOS) in submandibular glands of NOD mice in relation to parotid glands and unrelated tissues. Here we report that NOD mice with incipient signs of secretory dysfunction presented a marked decrease in basal and vasoactive intestinal peptide (VIP)-stimulated NOS activity and a differential expression of NOS I in submandibular glands compared to control BALB/c mice. Similar alterations in NOS I were found in parotid glands but not in brain or spleen of NOD mice. No differences between NOD and controls appeared in NOS II and NOS III expression in any of the tissues studied.

Amylases↗

[Keratin filaments in epithelial cells of the excretory ducts of rabbit submandibular glands--an immunohistochemical and ultraimmunohistochemical study].

To clarify the roles of various keratin proteins, the distributions of eight keratin intermediate filament proteins (keratins 7, 8, 10, 13, 14, 18, 19 and 20) in the epithelial cells of the excretory ducts of rabbit submandibular glands were studied immunohistochemically and ultraimmunohistochemically. The epithelia of excretory ducts were composed of columnar cells and basal cells. In the columnar cells, intermediate filaments formed a network at the apical area, that is, an apex network connected with desmosomes. Keratins 7, 18 and 20 were detected in the upper layer of the network and keratins 8, 18 and 20 in the lower layer. The intermediate filaments containing keratin 7 were also connected with hemidesmosomes on the basal side. Keratins 7, 18 and 20 were found throughout the entire cytoplasm of the columnar cells. Keratins 8 and 14 were expressed near the nucleus, forming a ring-like structure around the Golgi apparatus in the columnar cells. In the basal cells, by contrast, the intermediate filaments were concentrated around the nucleus, forming a juxtanuclear network which contained keratin 10. Keratin 13 was detected between the juxtanuclear network and the cell membrane, and was connected with both desmosomes and hemidesmosomes. Kratin 7 filaments were contained throughout the entire cytoplasm of the basal cells. These results suggested that different functional subsets of keratin filaments could be distinguished in the epithelial cells of the excretory ducts of the submandibular glands. In the columnar cells, keratins 7, 8, 18 and 20 play a role in cell-cell contact or cell-matrix contact, and both keratins 8 and 14 seem to be involved in the structure of the Golgi apparatus. In the basal cells, keratin 10 may serve to position and anchor the nucleus within the cell, and keratin 13 plays a role in cell-cell and cell-matrix contacts.

Animals↗

Kallikrein-like esteroprotease activity and kininase activity in submandibular gland from a mutant mouse with human cystic fibrosis like alterations.

The submandibular gland of cri/cri and control mice were compared for their activity of glandular kallikrein like esteroprotease and kininase. Esteroprotease activity is significantly reduced in cri/cri mice with respect to control, with an increased kininase activity in cri/cri mice. Since previous work showed an electrolyte abnormality in the salivary glands of this mutant mouse (1) a possible relationship between this alteration with the low activity of cellular esteroprotease and the high kininase activity is suggested.

Animals↗

Changes in glycoconjugates in rat submandibular gland after chronic treatment with reserpine and isoproterenol.

Chronic treatment of rats with reserpine, isoproterenol, or a combination of these two agents has been suggested as a means to produce an experimental animal model for the chronic exocrinopathy cystic fibrosis. The effect of these treatments on glycoconjugate distribution in rat submandibular gland acinar cells was investigated by quantitative lectin cytochemistry. Significant changes in wheat-germ agglutinin (WGA), soy bean agglutinin (SBA) and concanavalin A (Con A) binding sites in the mucus granules were observed, but peanut agglutinin (PNA) binding was not significantly affected. The quantitative changes in glycoconjugates in the acinar cells of the submandibular gland could be a possible explanation for the increased binding of calcium by the intracellular mucus noted in previous studies on these animal models.

Animals↗

Filtration coefficient and osmotic reflection coefficient to albumin in rabbit submandibular gland capillaries.

1. The isolated perfused submandibular salivary gland of the rabbit has been used in order to make estimates of the filtration coefficient (Kf) and reflection coefficient (sigma d) of the capillary wall to albumin. 2. An isogravimetric preparation was used and in paired experiments the value for Kf obtained in glands perfused with albumin-Krebs solution, 0.96 +/- 0.086 (mean +/- S.E. of mean) ml min-1 mmHg-1 100 g-1, was not significantly different from that in blood-perfused glands, 0.90 +/- 0.15. 3. On analysing the data for reflection coefficient, it was concluded that the above values underestimated Kf by about 30%; using corrected values for Kf, osmotic reflection coefficients were determined from the weight changes following a sudden change in the oncotic pressure of the perfusate. The value for sigma d to albumin lay between 0.79 and 1.0, the lower value being obtained after the Kf correction. 4. The high hydraulic conductivity, combined with sieving properties comparable to those in continuous capillaries, is consistent with other data on fenestrated capillaries. 5. Finally, it was observed that, while the Kf value calculated from the initial flux rate was similar whether measured during fluid efflux from or influx into the microvasculature, on returning to the initial conditions after raising osmotic pressure, efflux was now more rapid than influx. This phenomenon is discussed in relation to readjustment of Starling forces and the possible existence of an asymmetric double membrane in the capillary, interstitium system and cells of the salivary gland.

Albumins↗

Ciliated cells in the main excretory duct of the submandibular gland in obstructive sialadenitis: a SEM and TEM study.

The epithelium of the main excretory duct of the submandibular gland in five cases of obstructive sialadenitis was studied by TEM and SEM. In three cases we found a considerable increase in goblet and ciliated cells. As well as normal cilia, different kinds of ciliary anomalies, especially compound cilia, were noticed. Images of centriole formation and of ciliogenesis, both normal and abnormal, were also observed. Several hypotheses concerning the mode of formation of the compound cilia are discussed.

Adult↗

Intercellular contacts at the epithelial-mesenchymal interface of the developing rat submandibular gland in vitro.

An ultrastructural study of the development of the rat submandibular gland (SMG) analage in vitro was undertaken to determine if epithelial-mesenchymal and epithelial-nerve contacts were integral events in the differentiation of the gland in vitro as they are in vivo. SMG rudiments were removed at the stalk-bulb stage (15 days in utero) and cultured for 6 days on a millipore filter in supplemented McCoy's 5A media. Rudiments were taken at daily intervals, fixed and processed for electron microscopy. The overall development of the explanted rudiments closely paralleled their maturation in vivo although cultured glands lagged 24-36 h behind their normal counterparts. Direct epithelial-mesenchymal contacts were seen after the morphogenetic patterning of the gland had been established but prior to functional differentiation of the rudiment. Epithelial-nerve contacts were not seen although healthy axons were seen in the stroma throughout the culture period. The study indicates that epithelial-nerve contacts are probably not required for morphogenesis of cytodifferentiation of the rat SMG. However, direct epithelial-mesenchymal contacts appear to be an integral part of the developmental sequence of the rat SMG.

Animals↗

The role of tissue mast cells in exocrine secretion: studies in the submandibular gland of the cat.

Histamine was found to be stored in the submandibular gland of the cat mainly in the mast cells. The amine is released from mast cells by compound 48/80 and pilocarpine. This was demonstrated in the in vivo and in vitro experiments and by histological examination. During the physiological stimulation of the gland, via the electrical stimulation of the chorda tympani nerve, significant changes of histamine content were not found. This could be explained by the increased synthesis of histamine during physiological stimulation. The physiological role of histamine in salivary secretion was demonstrated. The similarity between the roles of histamine, mast cells and chromaffine-like cells in salivary and gastric secretion is discussed.

Animals↗