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Specification of cell fates within the salivary gland primordium.

The Drosophila salivary gland is a simple tubular organ derived from a contiguous epithelial primordium, which is established by the activities of the homeodomain-containing proteins Sex combs reduced (SCR), Extradenticle (EXD), and Homothorax (HTH). EGF signaling along the ventral midline specifies the salivary duct fate for cells in the center of the primordium, while cells farther away from the source of EGF signal adopt a secretory cell fate. EGF signaling works, at least in part, by repressing expression of secretory cell genes in the duct primordium, including fork head (fkh), which encodes a winged-helix transcription factor. FKH, in turn, represses trachealess (trh), a duct-specific gene initially expressed throughout the salivary gland primordium. trh encodes a basic helix-loop-helix PAS-domain containing transcription factor that has been proposed to specify the salivary duct fate. In conflict with this model, we find that three genes, dead ringer (dri), Serrate (Ser), and trh itself, are expressed in the duct independently of trh. Expression of all three duct genes is repressed in the secretory cells by FKH. We also show that SER in the duct cells signals to the adjacent secretory cells to specify a third cell type, the imaginal ring cells. Thus, localized EGF- and Notch-signaling transform a uniform epithelial sheet into three distinct cell types. In addition, Ser directs formation of actin rings in the salivary duct.

Animals↗

Effects of retinoic acid on morphological features and biological markers of a neoplastic human salivary intercalated duct cell line in culture.

Retinoic acid has marked effects on the growth, morphological features, and biological markers of a neoplastic human salivary intercalated duct cell clone in culture, whereas the cell clone was not affected by other retinoids such as retinol and retinal. A cell clone with ultrastructure and biological markers specific to the intercalated duct cells of human salivary glands was cultivated in the presence of retinoic acid. Major alterations, such as expression of tonofilaments, Mr 68,000 cytokeratin, and involucrin, were observed in those cells with a phenotype similar to that of keratinizing squamous cells. In addition, the coexpression of Mr 68,000 cytokeratin and carcinoembryonic antigen in these altered cells was found. Both the anchorage-independent and anchorage-dependent growths were markedly suppressed in the presence of retinoic acid. After the removal of retinoic acid from the culture, the treated cells returned rapidly to the phenotype of the untreated cells. These findings indicate that reversible differentiation into the keratinizing squamous cells of a neoplastic human salivary intercalated duct cell clone occurs in growth medium containing retinoic acid.

Adenocarcinoma↗

Endothelial constitutive nitric oxide synthase (ecNOS) localization in normal and neoplastic salivary tissue.

BACKGROUND: Nitric oxide (NO.) has been implicated in the process of carcinogenesis in various organs. This study was designed to investigate the expression of endothelial constitutive nitric oxide synthase (ecNOS) in normal and neoplastic salivary tissues. METHODS: Paraffin-embedded tissue from 48 salivary tumors and adjacent non-neoplastic tissue was immunohistochemically evaluated for both frequency (percentage) and intensity (1-4+) of staining using a commercially available anti-ecNOS monoclonal antibody. RESULTS: Expression of ecNOS was predominantly localized to vascular endothelium, skeletal muscle, and to salivary duct luminal epithelium in normal salivary tissue (n = 37). All salivary tumors demonstrated at least 1 + cytoplasmic staining for ecNOS without apparent correlation to most clinical parameters. A tendency toward increased frequency and intensity of ecNOS expression in oncocytic cells, relative to cells with myoepithelial or acinar differentiation, was noted. CONCLUSIONS: Expression of ecNOS is localized to the luminal cells of normal salivary ducts. Limited expression of ecNOS was found in all the salivary gland tumors examined. This suggests a common histogenesis for this diverse group of tumors, which may reflect different degrees of differentiation toward luminal duct epithelium. The possible role of ecNOS and NO. in salivary gland carcinogenesis is intriguing and warrants further study.

Adult↗

Hybrid carcinomas of the salivary glands: report of nine cases with a clinicopathologic, immunohistochemical, and p53 gene alteration analysis.

Hybrid carcinomas of the salivary gland are a recently defined and rare tumor entity, consisting of two histologically distinct types of carcinoma within the same topographic area. In this study, we examined nine such cases, which mainly arose in the parotid gland (seven cases), with an additional one each from submandibular and lacrimal glands, and analyzed their clinicopathologic profiles, including immunohistochemical features and p53 gene alterations. The prevalence of hybrid carcinomas was 0.4% among the 1863 cases of parotid gland tumors in our series. The nine patients comprised five men and four women, ranging in age from 40 to 81 years (mean, 62 y). Tumor size ranged from 2 to 10 cm (mean, 4.2 cm). Of the seven patients who were followed up, two were alive with disease and five were alive with no evidence of disease, although the follow-up period was short. Three cases had cervical lymph nodal metastases. The combinations of carcinoma components in our hybrid carcinomas were as follows: epithelial-myoepithelial carcinoma and basal cell adenocarcinoma in two cases, epithelial-myoepithelial carcinoma and squamous cell carcinoma in one case, salivary duct carcinoma and adenoid cystic carcinoma in two cases, myoepithelial carcinoma and salivary duct carcinoma in one, acinic cell carcinoma and salivary duct carcinoma in one, and squamous cell carcinoma and salivary duct carcinoma in two. Although the proportion of each carcinoma component in a tumor mass varied from case to case, the minor component always represented >or= 10% of the area. Differences in cellular composition were studied by immunohistochemistry and electron microscopy. The Ki-67-labeling index apparently differed between the two carcinoma elements in five cases. Diffusely positive p53 immunoreactivity was observed in four cases, restricted to the more aggressive component in each pair. Furthermore, p53 gene alteration analysis of these p53-positive cases revealed that all and three cases demonstrated loss of heterozygosity at p53 microsatellite loci and p53 gene point mutations, respectively, which were detected only in the p53-immunoreactive carcinoma component. Therefore, there is the possibility that such molecular-genetic events take an integral part for inducing the transformation from histologically lower to higher grade tumor during the hybrid carcinoma genesis of the salivary glands.

Adult↗

Analysis of chromosome 9p21 deletion and p16 gene mutation in salivary gland carcinomas.

The chromosomal locus 9p21 contains the p16(INK4a/CDKN2/MTS1) tumor suppressor gene that has been implicated in a variety of tumor types, including carcinomas of the head and neck, esophagus, and pancreas. To determine whether the loss of this gene is involved in salivary gland cancers, 35 carcinomas and paired nonneoplastic specimens were analyzed for loss of heterozygosity (LOH) of polymorphic genetic markers located in the region of interest. Five types of salivary gland tumors were studied: mucoepidermoid carcinoma, salivary duct carcinoma, adenoid cystic carcinoma, acinic cell carcinoma, and polymorphous low-grade adenocarcinoma. Seven of 9 salivary duct carcinomas showed LOH of 1 or more polymorphic markers. In 1 case of salivary duct carcinoma with LOH, we confirmed a deletion of bp 240-254 within exon 2. In addition, another salivary duct carcinoma showed a homozygous deletion of p16 in differential polymerase chain reaction analysis. Loss of heterozygosity was found in 1 of 10 adenoid cystic carcinomas and 1 of 8 mucoepidermoid carcinomas and was absent in the remaining subtypes. No mutations in exon 1 or exon 2 or homozygous deletion of p16 were found in these 2 particular neoplasms with LOH. These results suggest that inactivation of p16 is important in the development or progression of at least some salivary duct carcinomas, but we found no evidence that its alteration plays a role in the other subtypes examined.

Adenocarcinoma↗

Clinical application of magnetic resonance sialographic 3-dimensional reconstruction imaging and magnetic resonance virtual endoscopy for salivary gland duct analysis.

PURPOSE: In a previous study, we showed that the new and fast sequence 3-dimensional (3D)-fast asymmetric spin-echo sequencing could be applied in magnetic resonance (MR) sialographic 3D reconstruction imaging of the parotid gland ducts and in producing virtual endoscopic views of the parotid gland with MR data. MATERIALS AND METHODS: In the present study, we examined the clinical application of these MR sialographic 3D reconstruction imaging and virtual endoscopy of the salivary gland ducts using MR data sets with 3D-fast asymmetric spin-echo sequencing. RESULTS: The MR sialographic 3D reconstruction images showed a complete view in the branch paths from all angles, and the MR virtual endoscopic views showed conditions in the endoluminal tracts of the large branches in 20 patients with salivary gland duct abnormalities, including Sjögren syndrome, cyst, tumor, sialadenitis, and salivary calculi. CONCLUSION: The clinical use of MR sialographic 3D reconstruction imaging and MR virtual endoscopy for salivary gland ducts may enhance understanding of the 3D relationship between the ducts and the surrounding tissue, as well as the endoluminal circumstance within ducts. Possible future applications abound, and further investigation in this field is expected.

Adolescent↗

Effect of epidermal growth factor on the cellular proliferation and phenotype of a neoplastic human salivary intercalated duct cell line or its derivatives.

A neoplastic human salivary intercalated duct cell line (HSG) and its derivatives, HSG with a myoepithelial cell phenotype (HSG-AZA1) and HSG with an acinar cell phenotype (HSG-AZA3), which were induced by 5-azacytidine treatment of HSG cells, were cultivated in the presence of epidermal growth factor (EGF). Morphological changes occurred; cells that were spindle shaped or stellate and had long cytoplasmic processes appeared in all of the treated cells. Major alterations, such as expression of neuron-specific enolase, neurofilaments, or synaptophysin as well as formation of neurite-like structures densely packed with microfibrils and microtubules, were observed in these cells with a phenotype similar to that of neuron-like cells. Both the anchorage-independent and anchorage-dependent growths of the treated HSG cells and only anchorage-dependent growth of the treated HSG-AZA1 cells were suppressed, whereas the anchorage-dependent growth of the treated HSG-AZA3 cells was enhanced according to the increasing concentrations of EGF. In addition, it has been found by statistical analysis that the growth of HSG cells in culture is suppressed and that of HSG-AZA3 cells in culture is stimulated according to the increasing concentrations of EGF added, whereas the growth of cultured HSG-AZA1 cells is hardly affected by EGF treatment. The HSG, HSG-AZA1, and HSG-AZA3 cell had 1.8 x 10(5), 1.83 x 10(5), and 1.34 x 10(5) EGF receptors, respectively. The amounts of the EGF internalized into cells were larger in HSG-AZA3 cells than in HSG or HSG-AZA1 cells. These findings indicate that the conversion of HSG, HSG-AZA1, and HSG-AZA3 cells into neuron-like cells occurs in growth medium containing EGF, with a concomitant modulation of the growth potentials of the cells which were examined in a different manner.

Amylases↗

Role of mandibular striated duct in salivary excretion of urea in dogs.

The role of mandibular striated duct in salivary excretion of endogenous urea was studied by a technique of retrograde ductal injection of HgCl2 and a stop flow analysis. (1) It was verified in this study that treatment with HgCl2 markedly inhibited Na+ reabsorption in the striated duct so that water reabsorption driven by the Na+ reabsorption would be inhibited substantially. (2) The saliva/plasma concentration ratios (S/P ratios) of urea in HgCl2-treated saliva (about 0.71) were significantly higher than those in untreated saliva (about 0.37). (3) The excretion rate of urea in HgCl2-treated saliva (about 0.02 mg/min/kg) was also significantly higher than those in untreated saliva (about 0.01 mg/min/kg) despite the same flow rate for both saliva samples. Furthermore, (4) The S/P ratios of urea in the second (0.25-0.50 ml) poststop-flow saliva (about 0.65) were significantly higher than those in prestop-flow saliva (about 0.35). These results suggest a possibility that salivary urea is reabsorbed in the dog mandibular striated duct and that the enhancement of the S/P ratio as well as the salivary level of urea by the pre-treatment with HgCl2 may be due to the inhibition of urea reabsorption.

Animals↗

Salivary gland duct involvement in oral epithelial dysplasia and squamous cell carcinoma.

The clinical implications and prognostic significance of oral dysplastic or cancerous epithelium involving salivary gland ducts have not been previously investigated. Screened routine tissue sections of 1216 cases of oral epithelial dysplasias and squamous cell carcinomas revealed 26 examples (2.14%) that exhibited unequivocal ductal involvement. Ductal involvement was more likely to occur in floor of mouth lesions and in lesions exhibiting severe dysplasia or carcinoma in situ. Clinical follow-up on 23 cases showed that the recurrence rate of the preinvasive lesions that exhibited ductal involvement was equal to that of the squamous cell carcinomas. The depth of ductal dysplasia did not correlate with recurrence rate. These results suggest that the involvement of salivary gland ducts by oral epithelial dysplasias and carcinomas in situ is an uncommon but significant finding. Surgical stripping or ablation of such lesions should extend at least 3 mm below the surface to ensure eradication of these reservoirs of dysplastic cells.

Aged↗

Virtual endoscopic view of salivary gland ducts using MR sialography data from three dimension fast asymmetric spin-echo (3D-FASE) sequences: a preliminary study.

OBJECTIVES: We performed magnetic resonance (MR) sialography of parotid gland and/or submandibular gland ducts using three-dimensional fast asymmetric spin-echo (3D-FASE) sequencing. The objective was to make three-dimensional (3D) reconstruction images and virtual endoscopic views of the parotid gland ducts using MR sialography data sets of 3D-FASE sequences. METHODS: We reviewed the MR sialography data sets with 3D-FASE sequencing of 10 control volunteers and six patients. Three-dimensional reconstruction images and virtual endoscopic views of the parotid gland and/or submandibular gland ducts were generated with maximum intensity projection (MIP), shaded surface display (SSD), and volume rendering techniques (VRT). RESULTS: The main parotid gland and/or submandibular gland ducts, large branches within the glands, and small branches were fairly well defined in a very short acquisition time on MR sialographic images with 3D-FASE sequencing in nine of the 10 healthy volunteers. The 3D-reconstruction images of the parotid gland ducts and/or submandibular gland ducts showed the entire length of the branch paths and complete images from all angles, and the virtual endoscopic views showed the endoluminal tracts of the main ducts and the large branches in nine. In the patient with Sjogren's syndrome, chronic sialoadenitis, and salivary calculi in the Wharton ducts, the MR sialographic images showed diffuse areas of punctate high signal intensity, dilatation of Stensen's duct, or stones of Wharton's duct, respectively. Furthermore, the 3D-reconstruction images of the salivary gland ducts showed the stenoses and stones in the branch paths and complete images from all angles, and the virtual endoscopic views showed the stenoses and stones in the endoluminal tracts of the main and large branches. CONCLUSIONS: Our initial experience showed that virtual MR endoscopy could be performed to observe the endoluminal tracts of parotid and submandibular glands. The clinical use of the virtual MR endoscopy for salivary gland ducts has not been established yet. Future applications of the 3D-reconstruction images and virtual endoscopic views using MR sialography data sets of 3D-FASE sequences are very attractive and further expansion of this field is expected.

Adolescent↗

Duct carcinoma of the minor salivary glands: a case report.

A tumour was removed from the oral vestibule of a 62-year-old woman. It showed histological features of both salivary duct carcinoma and papillary adenocarcinoma. The possibility that these entities might be but varieties of duct adenocarcinoma is suggested.

Carcinoma, Intraductal, Noninfiltrating↗

Expression of nitric oxide synthase in pleomorphic adenomas of the parotid.

The actions of nitric oxide (NO) in the pathology of solid tumours are complicated and many are poorly understood because NO has both inhibitory and tumour-promoting activities. In the current study we aimed to find out immunohistochemically whether the expression of both the inducible (iNOS) and endothelial (eNOS) forms of the enzyme nitric oxide synthase (NOS) were changed in pleomorphic adenomas of the parotid compared with normal salivary tissue. There was a significant difference in staining for iNOS between the tumour and normal salivary tissue, with tumour epithelial cells being stained in 29 cases of the 30 cases studied (P< 0.0001). The luminal cells of the salivary ducts also stained, but not the normal salivary tissue. Immunohistochemistry for the eNOS isoenzyme showed moderate staining of the tumour epithelium in only three specimens. There was also mild staining in the salivary duct cells of the normal glandular tissue and in endothelium of blood vessels in both tumour and normal glandular tissue in the same 29 cases.

Adenoma, Pleomorphic↗

[Surgical treatment of salivary incontinence in cerebral palsy].

Swallowing dysfunction is the main cause of drooling in cerebral palsy. Medical treatment is inefficient. Surgical treatment involves neurectomy, translocation of the salivary duct or salivary gland resection. Following this, reeducation is advisable. Indications are based on the degree of drooling and on the degree of histories mental retardation. The authors present 2 case histories of drooling in patients with cerebral palsy treated by submandibular gland resection and parotid duct ligation.

Adolescent↗

Cellular expression of Noc2, a Rab effector protein, in endocrine and exocrine tissues in the mouse.

Noc2 is a Rab effector which participates in regulated exocytosis. It is expressed abundantly in endocrine cells but at low levels in exocrine tissues. Noc2-deficient mice, however, exhibit marked accumulation of secretory granules in exocrine cells rather than endocrine cells. In the present study, we investigated localization of Noc2 immunohistochemically in various endocrine and exocrine tissues in normal mice. Western blotting detected a Noc2-immunoreactive band of 38 kDa in isolated pancreatic islets, the adrenal gland, pituitary gland, and thyroid gland. Immunostaining for Noc2 labeled endocrine cells in the adrenal medulla and adenohypophysis, pancreatic islet cells, thyroid parafollicular cells, and gut endocrine cells, supporting the notion that Noc2 is a Rab effector protein shared by amine/peptide-secreting endocrine cells. Besides endocrine tissues, granular ducts in salivary glands contained Noc2. Although immunostaining failed to detect Noc2 in acinar cells of all exocrine glands examined, reverse transcriptase-polymerase chain reaction analysis detected the mRNA expression in exocrine pancreas. Ultrastructurally, Noc2 immunoreactivity was associated with the limiting membrane of granules in both pancreatic endocrine and salivary duct exocrine cells. The cellular and subcellular localizations of Noc2 should yield key information on its functional significance as well as account for the phenotype in Noc2-deficient mice.

Adaptor Proteins, Signal Transducing↗

Keratin 19-like immunoreactivity in receptor cells of mammalian taste buds.

Three monoclonal antibodies, 4.62, LP2K and 170.2.14, were used to evaluate keratin 19-like immunoreactivity in gustatory epithelia. Keratin 19-like immunoreactivity was restricted to the intragemmal cells for all types of mammalian taste buds examined. These taste buds included fungiform, foliate and vallate taste buds in rat, gerbil and rabbit, and nasopalatine, epiglottal and palatine taste buds in rat. There was no keratin 19-like immunoreactivity in basal cells or in perigemmal cells lateral to the immunoreactive taste receptor cells. Denervation of the rat vallate papilla eliminated all taste buds, as well as all immunoreactive taste cells. That the immunoreactive material in the taste cells was keratin 19 was supported by the comparable staining of rat taste buds with each of three monoclonal antibodies specific for keratin 19. Furthermore, as predicted, these antibodies selectively stained luminal cells of rat bile ducts, bladder, salivary ducts, trachea, ureter and uterus. It was concluded that monoclonal antibodies against keratin 19 can usefully distinguish intragemmal taste receptor cells from keratinocytes, and from the perigemmal and basal cells of gustatory epithelia. Anti-keratin 19 antibodies may serve to identify differentiated taste cells in gustatory epithelia undergoing taste bud development, renewal, degeneration or regeneration.

Animals↗

Human monoclonal antibodies directed against melanoma tumor-associated antigens.

To analyze the humoral immune response to melanoma, human-mouse hybridomas were generated by the fusions of regional lymph node lymphocytes of patients with the mouse myeloma cell line M5. Six stable hybridomas were cloned from six separate lymphocyte parents obtained from three patients. Ascites were obtained from nude mice after i.p. injection with cultured hybridoma cells. The monoclonal antibodies, four immunoglobulin Gs and two pentameric immunoglobulin Ms, were partially purified to remove mouse immunoglobulin and then conjugated to biotin for immunocytochemical and immunohistochemical studies. With the avidin:biotin:peroxidase complex method to detect and amplify binding by the biotin-conjugated human monoclonal antibodies, we found the six antibodies to be reactive against cytoplasmic determinants in five short-term melanoma cultures and formalin-fixed paraffin-embedded melanoma tumors from four patients. The antigenic target of the antibodies identified was not carcinoembryonic antigen. Two antibodies, 2-139-1 and 6-26-3, were studied in more detail. Each stained 25 of 25 specimens of melanomas. Little or no reactivity was detected against fixed sections of normal skin, which included tissues such as epidermis, dermis, monocytes, lymphocytes, and vascular endothelium. More striking was the absence of binding to melanocytes in the basal layer of the skin or to pigmented nevus cells. Both antibodies showed cross-reactivity against other tumors, in particular colonic and prostatic carcinomas. In the normal colon, reactivity was restricted to the surface of the columnar epithelium; no reactivity was detected against normal prostatic epithelium. Reactivity was also not observed against liver and lung. However, the epithelia of the renal tubules, pancreatic ducts, and salivary ducts were all reactive. These human monoclonal antibodies identify cytoplasmic melanoma-associated tumor antigens that appear different from the membrane antigens defined by serological approaches and by most mouse monoclonal antibodies.

Antibodies, Monoclonal↗