Prostate-specific antigen: An unfamiliar protein in the human salivary glands.

OBJECTIVES: The presence of prostate-specific antigen (PSA) in saliva and salivary glands has been reported. Nevertheless, its release pathway in these glands remains to be elucidated. Here, we showed PSA subcellular distribution focusing on its plausible route in human salivary parenchyma. MATERIALS AND METHODS: Sections of parotid and submandibular glands were subjected to the immunohistochemical demonstration of PSA by the streptavidin-biotin method revealed by alkaline phosphatase. Moreover, ultrathin sections were collected on nickel grids and processed for immunocytochemical analysis, to visualize the intracellular distribution pattern of PSA through the observation by transmission electron microscopy. RESULTS: By immunohistochemistry, in both parotid and submandibular glands PSA expression was detected in serous secretory acini and striated ducts. By immunocytochemistry, immunoreactivity was retrieved in the cytoplasmic compartment of acinar and ductal cells, often associated with small cytoplasmic vesicles. PSA labeling appeared also on rough endoplasmic reticulum and in the acini's lumen. A negligible PSA labeling appeared in most of the secretory granules of both glands. CONCLUSIONS: Our findings clearly support that human parotid and submandibular glands are involved in PSA secretion. Moreover, based on the immunoreactivity pattern, its release in oral cavity would probably occur by minor regulated secretory or constitutive-like secretory pathways.

Histopathological and biochemical alterations of the parotid gland induced by experimental hypothyroidism in adult male rats and the possible therapeutic effect of Nigella sativa oil.

Thyroid hormones are essential for metabolic rate regulation and play a role on the integrity of the salivary glands. Nigella sativa is a widely used plant in medicine. This study aimed to evaluate the effect of Nigella sativa oil (NSO) on the hypothyroidism-induced parotid gland pathological alterations. Rats were divided into four groups: control group, hypothyroid group: received daily oral carbimazole for 3 weeks, hypothyroid-NSO group: NSO was orally given for 4 weeks after hypothyroidism induction and NSO group: administrated NSO only for 4 weeks. After 7 weeks, all rats were sacrificed, serum thyroid hormones were estimated, and parotid glands were assessed by histopathological, immunohistochemical, ultrastructural and morphometric analyses. Hypothyroid group showed a significant decrease in thyroid hormones with increase in thyroid-stimulating hormone (TSH) levels and decrease in body and parotid weights compared to the control rats that were improved with NSO treatment. Sections of the hypothyroid group showed fibrosis, acinar cytoplasmic vacuolations, vascular congestion, ductal dilatation, wide intercellular canaliculi, nuclear pyknosis and decreased number of secretory granules. Also, there were decreased B-cell lymphoma 2 (Bcl-2) and increased p53, Bcl-2 Associated X (Bax) and alpha-smooth muscle actin (α-SMA) immune-expressions; with decreased Bax/ Bcl-2 ratio that all were attenuated by NSO. NSO ameliorates hypothyroidism-induced parotid changes by altering p53, Bax and Bcl-2 pathway.

Localization of phosphatidylinositol 4-phosphate 5-kinase (PIP5K) α, ß, γ in the three major salivary glands in situ of mice and their response to ß-adrenoceptor stimulation.

Phosphatidylinositol 4-phosphate 5-kinase (PIP5K), which is composed of three isozymes (α, ß and γ), catalyzes the production of phosphatidylinositol bisphosphate (PIP2). This phospholipid functions in membrane trafficking, as an anchor for actin cytoskeletons and as a regulator of intramembranous channels/transporters. It is also a precursor of such second messengers as diacylglycerol, inositol triphosphate and phosphatidylinositol (3,4,5)-triphosphate. In the present study, the expression and localization of endogenous PIP5Ks were examined in the three major salivary glands of young adult mice in situ. In western blotting of normal control glands, immunoreactive bands for individual PIP5Ks were detectable, with the highest density in the parotid gland and the weakest density in the submandibular gland. In immuno-light microscopy under non-stimulated condition, weak immunoreactivity for PIP5Kα was confined to the apical plasmalemma in parotid, but not sublingual or submandibular, acinar cells. Immunoreactivity for PIP5Kß was weak to moderate and confined to ductal cells but not acinar cells, whereas that for PIP5Kγ was selectively and intensely detected in myoepithelial cells but not acinar cells, and it was weak in ductal cells in the three glands. In western blot of the parotid gland stimulated by isoproterenol, a ß-adrenoceptor agonist, no changes were seen in the intensity of immunoreactive bands for any of the PIP5Ks. In contrast, in immuno-light microscopy, the apical immunoreactivity for PIP5Kα in parotid acinar cells was transiently and distinctly increased after the stimulation. The increased immunoreactivity was ultrastructurally localized on most apical microvilli and along contiguous plasma membrane, where membranous invaginations of various shapes and small vesicles were frequently found. It was thus suggested that PIP5Kα is involved in post-exocytotic membrane dynamics via microvillous membranes. The present finding further suggests that each of the three isoforms of PIP5K functions through its product PIP2 discretely in different cells of the glands to regulate saliva secretion.

Bone marrow-derived mesenchymal stem cells ameliorate parotid injury in ovariectomized rats.

BACKGROUND AIMS: Parotid hypofunction causes life-disrupting effects, and there are no effective medications for xerostomia. We hypothesized that mesenchymal stem cells (MSCs) have repairing effects on parotid glands of ovariectomized (OVX) rats. METHODS: Forty-five adult female rats were divided into three equal groups: group I (Control group), group II (OVX-group) and group III (OVX rats that received MSCs at 4 and 8 weeks post-ovariectomy). At 12 weeks post-ovariectomy, histological (Masson's trichrome and periodic acid-Schiff with alcian blue stains), immunohistochemical (caspase-3 and CD44) and morphometric studies and salivary flow rate and saliva pH determination were carried out. RESULTS: Histologically, the OVX group displayed numerous irregular vacuolated acini, thickened septa with marked cellular infiltration and vascular congestion. Degenerated organelles and few or irregular secretory granules with a different density were observed. Caspase-3-positive cells were highly expressed. MSC-treated glands exhibited a considerable degree of preservation of glandular architecture with numerous CD44-expressing and few caspase-3-expressing cells. Significant decrease of the salivary flow rate in the OVX group was detected, which reverted to normal levels in group III. CONCLUSIONS: MSCs ameliorated the damaging effects of ovariectomy on the parotid glands.

Developmental changes in the parotid salivary gland of prenatal buffalo: an ultrastructural study / Cambios en el desarrollo de la glándula salival de la parótida de búfalo prenatal: un estudio ultraestructural

SUMMARY: The present study was undertaken to elucidate ultrastructural changes in development of parotid salivary gland of buffalo during different stages of prenatal life. The ultrastructural studies revealed that the cytoplasm of acinar cells was filled with mitochondria, rough endoplasmic reticulum and Golgi complex in mid and late foetal age groups. Medium electron dense secretory granules first appeared in the acinar cells of parotid gland at 30 cm CVRL (141st day). However, at 49.5 cm CVRL (185th day) two types of electron dense granules were identified on basis of granule density viz., dark and light. The dark electron dense granules were more in number, whereas light granules were comparatively less having electron lucent content within them was identified. The mean diameter of dark and light granules was measured about 0.45±0.1 µm and 0.30±0.1 µm, respectively, which showed that the dark granules were comparatively larger in size. The secretory granules were increased in number during the late foetal age group. The myoepithelial cells were located at the base of the acinar cells as well as intercalated and striated ducts, and were stellate in shape. The ultrastructure of myoepithelial cell revealed parallel stream of myofilaments in the cytoplasm and its processes. Lipofuscin pigments were also observed in between the acinar cells of parotid gland.

RESUMEN: El estudio se realizó para elucidar los cambios ultraestructurales en el desarrollo de la parótida del búfalo durante las diferentes etapas de la vida prenatal. Los estudios ultraestructurales revelaron que el citoplasma de las células acinares estaba saturado de mitocondrias, de retículo endoplasmático rugoso y Complexo golgiensis en las edades fetal media y tardía. Se observó un número mayor de gránulos oscuros densos de electrones, mientras que los gránulos ligeros fueron comparativamente menor en número con contenido de electrones. El diámetro medio de gránulos oscuros y ligeros se midió aproximadamente 0,45 / pm 0,1 / mu m y 0,30 / pm 0,1 / mu m, respectivamente, lo que mostró que los gránulos oscuros eran comparativamente mayores en tamaño. Los gránulos secretores aumentaron en número durante el último grupo de edad fetal. Las células mioepiteliales se localizaron en la base de las células acinares, así como en conductos intercalados y estriados, y tenían una forma estrellada. La ultraestructura de las células mioepiteliales reveló una corriente paralela de miofilamentos en el citoplasma y sus procesos. También se observaron pigmentos de lipofuscina entre las células acinares de la glándula parótida.

Análisis morfométrico ultraestructural de células normales de glándula parótida de rata / Ultrastructural morphometric analysis from normal parotid gland cells of rat

A partir de 10 ratas hembras con un peso aproximado de 250 g y 4 meses de vida, fueron obtenidas quirúrgicamente muestras de glándula parótida las que se trataron con técnicas de microscopía electrónica de transmisión para posteriormente obtener microfotografías de células parotideas con aumentos finales de hasta 21300 X. En las citadas microfotografías se aplicaron técnicas morfométricas con el objetivo de cuantificar las fracciones volumétricas que los distintos componentes ocupan en estas células normales, describiendo de esta manera sus volúmenes y relacionándolos con la funcionalidad que desempeñan en esta célula normal. Se evaluaron las fracciones volumétricas pertenecientes a: citoplasma, núcleo, mitocondrias, retículo endoplasmático rugoso (RER), gránulos de zimógeno, eu y heterocromatina. De igual forma, se cuantificó las áreas celulares y nucleares. Contando con los datos numéricos producto de la evaluación morfométrica de sus componentes se podrá determinar el patrón de distribución de sus organelos y de funcionalidad de esta célula activa en la síntesis y secreción de proteínas representada por los gránulos de zimógeno de diastasa y diversas proteínas salivales.

From 10 female rats weighing approximately 250 g and aged 4 months, samples of parotid gland were obtained surgically which were treated with transmission electronic microscopy in order to obtain microphotographs with final increases of up to 21,300 X. Morphometric techniques were applied to these microphotographs to quantify the volumetric fractions that the different components occupy in these normal cells, thus describing their volumes and relating them to their functionality in this normal cell. Volumetric fractions were evaluated pertaining to: cytoplasm, nucleus, mitochondria, rough endoplasmic reticulum (RER), zymogen granules, eu and heterochromatin. Likewise, cell and nuclear areas were quantified. With the numerical data from the morphometric evaluation of its components, it was possible to determine the distribution pattern of the organelles and functionality of this cell active in protein synthesis and secretion represented by diastase zymogen granules and various salivary proteins.

Subcellular distribution of melatonin receptors in human parotid glands.

The hormone melatonin influences oral health through a variety of actions, such as anti-inflammatory, anti-oxidant, immunomodulatory and antitumour. Many of these melatonin functions are mediated by a family of membrane receptors expressed in the oral epithelium and salivary glands. Using immunoblotting and immunohistochemistry, recent studies have shown that the melatonin membrane receptors, MT1 and MT2, are present in rat and human salivary glands. To date, no investigation has dealt with the ultrastructural distribution of the melatonin receptors. This was the aim of the present study, using the immunogold method applied to the human parotid gland. Reactivity to MT1 and, with less intensity, to MT2 appeared in the secretory granules of acinar cells and in the cytoplasmic vesicles of both acinar and ductal cells. Plasma membranes were also stained, albeit slightly. The peculiar intracytoplasmic distribution of these receptors may indicate that there is an uptake/transport system for melatonin from the circulation into the saliva.

Is sonoelastography a helpful method for evaluation of parotid tumors?

Sonoelastography is a novel technique, useful in a noninvasive assessment of lesions in multiple organs. The aim of the study was to examine whether the combination of conventional ultrasonography (US) with sonoelastography might improve the reliability of parotid tumor evaluation. Fourty-three consecutive patients with parotid tumors were surgically treated at a single tertiary center at the Department of Otolaryngology, Head and Neck Surgery. The sample included 27 women and 16 men, aged 15-80 (the mean age = 54 years). The reference group constituted of 54 healthy volunteers. High resolution grayscale ultrasonography (US) was performed preoperatively using a 15 MHz linear array transducer. Elastograms (ES) were scored by the conventional Ueno 5-point scale from ES1 (blue-soft) to ES5 (the entire lesion and surrounding area shaded red-stiff). In addition, detailed stiffness values in kPa were collected. The group consisted of 33 patients with benign and 10 patients with malignant tumors. The mean stiffness value was 146.6 kPa in 10 malignant tumors (mostly ES4) and 88.7 kPa in 33 benign tumors (mostly ES2 and ES3). The differences in tissue stiffness between normal parotid parenchyma in the reference group and the mean value for all tumors in the examined group were statistically significant (p < 0.001), and so was the case with the differences between the benign and malignant tumors (p < 0.001). Low stiffness scores (ES1,2) were found in 2 malignant and 15 benign tumors while high scores (ES3,4) were found in 8 malignancies and 18 benign tumors. Sonoelastography overlapping elasticity to the grayscale images supports additional informations. Preferential selection of the lesions characterized by high stiffness (ES4) improves the differential diagnosis of parotid tumors but the large degree of uncertainty of this method should also be pointed out.

Radioprotective effect of lidocaine on function and ultrastructure of salivary glands receiving fractionated radiation.

PURPOSE: Radiation-induced xerostomia still represents a common side effect after radiotherapy for head-and-neck malignancies. The aim of the present study was to examine the radioprotective effect of lidocaine hydrochloride during fractionated radiation in an experimental animal model. METHODS AND MATERIALS: To evaluate the influence of different radiation doses on salivary gland function and the radioprotective effect of lidocaine, rabbits were irradiated with 15, 25, 30, and 35 Gy (equivalent doses in 2-Gy fractions equivalent to 24, 40, 48, and 56 Gy, respectively). Lidocaine hydrochloride (10 and 12 mg/kg) was administered before every radiation fraction in the treatment groups. Salivary gland function was assessed by flow sialometry and sialoscintigraphy, and the morphologic changes were evaluated using transmission electron microscopy. RESULTS: Functional impairment was first observed after 35 Gy and pretreatment with lidocaine improved radiation tolerance of both parotid and submandibular glands. The use of 12 mg/kg lidocaine was superior and displayed significant radioprotection with regard to flow sialometry and sialoscintigraphy. The ultrastructure was largely preserved after pretreatment with both lidocaine doses. CONCLUSIONS: Lidocaine represents an effective radioprotective agent and a promising approach for clinical application to avoid radiation-induced functional impairment of salivary glands.

Selective culture of different types of human parotid gland cells.

BACKGROUND: Advances in salivary gland tissue engineering can benefit patients diagnosed with xerostomia. Complexity of the gland explains the urgent demand for a reliable protocol to isolate and expand various gland cells that can be used for further study. METHODS: Three cells with different morphologies were isolated from the same human parotid glands using different culture medium systems and then were identified by the expressions from mRNA to the protein level. RESULTS: Among the 34 specimens, parotid gland acinar cells, myoepithelial cells, and fibroblasts expressing specific markers that belonged to individual cell types, were successfully isolated and expanded from 30 specimens without a complex mechanical process and expensive flow technique. CONCLUSION: The proposed protocol is simple with a high success rate to culture various gland cells, making it highly promising for use in future tissue engineering studies.

Ultrastructural assessment of the radioprotective effects of sodium selenite on parotid glands in rats.

The aim of this study was to assess the radioprotective effects of sodium selenite on parotid glands in rats by ultrastructural analysis of acinar cells. Four experimental groups were assessed; control, irradiated, selenium, and selenium/irradiated. The sodium selenite dose was 0.5 mg/kg, administered intraperitoneally 24 h before irradiation in the head and neck region with a single 15-Gy dose of gamma radiation. At 4, 8, 12, 48 and 72 h after irradiation, all animals were sacrificed and the parotid glands were removed. Radiation caused cellular changes from 4 h, and the organelles that presented the greatest alterations were the mitochondria and the secretion glands; nuclear alterations were also observed. Sodium selenite was found to have a radioprotective action, as the selenium/irradiated group presented with less damage when compared to the irradiated group. However, sodium selenite caused cellular alterations that were evident after 8 h, but with less damage when compared to those caused by radiation, which demonstrates a favorable risk-benefit for its use as a radioprotector. Thus, this research shows that sodium selenite has an effective radioprotective action in the parotid gland, which may contribute to the reduction of the adverse effects brought by the radiotherapy.

Expression and localization of prominin-1 in isoproterenol-treated rat parotid gland.

Prominin-1 (CD133) is a pentaspan cholesterol-binding membrane glycoprotein. Chronic treatment with isoproterenol, a beta-receptor agonist, induces several dramatic effects on salivary glands, such as enhanced DNA synthesis and proliferation of salivary acinar cells. In addition, the biosynthetic pathways of membrane lipids may be altered by the isoproterenol stimulation. The purpose of this study was to investigate the effect of isoproterenol administration on prominin-1 expression profiles in rat parotid gland by means of PCR and immunohistochemistry. Rats were chronically treated with the beta-adrenergic agonist for 1, 3, and 7 days. Our results showed that isoproterenol-treatment caused a down-regulation of prominin-1 on day 3 and 7 of treatment, as well as a differential immunostaining distribution pattern. This study suggests that isoproterenol-treatment may represent a useful tool to explore the molecular mechanisms involved in the synthesis and release of prominin-1. Such efforts could contribute to the development of diagnostic tools based on the detection of prominin-1 in biological fluids, such as saliva.

Amylase and cyclic amp receptor protein expression in human diabetic parotid glands.

BACKGROUND: Salivary dysfunction and oral disorders have been described in both type 1 and type 2 diabetes mellitus. However, the cellular and molecular consequences of diabetes on oral tissues remain to be ascertained. The purpose of this investigation was to study, by means of electron microscopy, the morphologic and molecular changes that occur in salivary glands during diabetes. METHODS: Biopsy samples of parotid glands were excised from non-diabetic and diabetic (type 1 and type 2) consenting patients and processed by standard methods for routine morphology and electron microscopic immunogold labeling. Specific antibodies were used to determine and quantify the expression of secretory proteins (alphaamylase and the regulatory subunit of type II protein kinase A). RESULTS: Morphologic changes in the diabetic samples included increased numbers of secretory granules, and alterations in internal granule structure. Quantitative analysis of immunogold labeling showed that labeling densities were variable among the parotid gland samples. In type 1 diabetes amylase expression was greater than in non-diabetic glands, whereas in type 2 diabetes it was not significantly changed. Expression of type II regulatory subunits was slightly, although not significantly, increased in acinar secretory granules of type 1 diabetic samples and was unchanged in type 2 diabetic samples. CONCLUSIONS: Our data show that diabetes elicits specific changes in secretory protein expression in human salivary glands, thus contributing to the altered oral environment and oral disease associated with diabetes.

Radiation-induced damage to microstructure of parotid gland: evaluation using high-resolution magnetic resonance imaging.

PURPOSE: To elucidate the radiation-induced damage to the microstructure of the parotid gland using high-resolution magnetic resonance imaging. METHODS AND MATERIALS: High-resolution magnetic resonance imaging of the parotid gland was performed before radiotherapy (RT) and during the RT period or < or =3 weeks after RT completion for 12 head-and-neck cancer patients using a 1.5-T scanner with a microscopy coil. The maximal cross-sectional area of the gland was evaluated, and changes in the internal architecture of the gland were assessed both visually and quantitatively. RESULTS: Magnetic resonance images were obtained at a median parotid gland dose of 36 Gy (range, 11-64). According to the quantitative analysis, the maximal cross-sectional area of the gland was reduced, the width of the main duct was narrowed, and the intensity ratio of the main duct lumen to background was significantly decreased after RT (p <.0001). According to the visual assessment, the width of the main duct tended to narrow and the contrast of the duct lumen tended to be decreased, but no significant differences were noted. The visibility of the duct branches was unclear in 10 patients (p = .039), and the septum became dense in 11 patients (p = .006) after RT. CONCLUSION: High-resolution magnetic resonance imaging is a noninvasive method of evaluating radiation-induced changes to the internal architecture of the parotid gland. Morphologic changes in the irradiated parotid gland were demonstrated during the RT course even when a relatively small dose was delivered to the gland.

Spontaneous oscillations in intracellular Ca(2+) concentration via purinergic receptors elicit transient cell swelling in rat parotid ducts.

Using multiphoton microscopy, we established that rat parotid ductal cells exhibit spontaneous oscillations in intracellular Ca(2+) concentration ([Ca(2+)](i)). These oscillatory Ca(2+) responses were observed during continuous perfusion with a physiological salt solution at 37 degrees C in the absence of calcium mobilizing agonist stimulation. The timing and patterns of these spontaneous Ca(2+) oscillations varied among individual ductal cells, and the average number of Ca(2+) responses in a single responding ductal cell was 2.1 in a 10-min recording period. High-speed scanning (0.6 s/image) revealed that most spontaneous elevations in [Ca(2+)](i) were initiated at the luminal side of ductal cells and spread toward the basal side within 2 s. Electron microscopic analysis after Ca(2+) imaging indicated that spontaneously oscillating ducts contained numerous granules at the luminal side, which is characteristic of granular ducts. These Ca(2+) oscillations were completely blocked by the purinergic receptor inhibitors 4-[[4-formyl-5-hydroxy-6-methyl-3-[(phosphonooxy)methyl]-2-pyridinyl]azo]-1,3-benzenedisulfonic acid (PPADS) and suramin but were not blocked by the muscarinic antagonist atropine or the alpha-adrenergic antagonist phentolamine. Simultaneous observation of fura-2 fluorescence and differential interference contrast (DIC) images showed that spontaneous elevations of [Ca(2+)](i) were well correlated with changes in shape of ductal cells. Using a plasma membrane fluorescence probe, SynaptoGreen C4, we found that the changes in DIC images reflected spontaneous cell swelling of ductal cells. Our findings present the possibility that purinergic receptors mediate spontaneous Ca(2+) oscillations in parotid ductal cells and regulate electrolyte reabsorption from the primary saliva in the resting state.

Adenosis poliquística esclerosante de la glándula parótida / Sclerosing polycystic adenosis of the parotid gland

La adenosis poliquística esclerosante de la parótida (APEP) es una enfermedad infrecuente, caracterizada por elementos histológicos inflamatorios reactivos de las glándulas salivales, pero la presencia de displasia y atiplas hace también posible pensar se trate de un pseudotumor o una neoplasia. La APEP posee además semejanzas histopatológicas con la enfermedad fibroquística de la mama y comparten receptores de progesterona y estrógenos en las células ductales. La edad promedio de ocurrencia de esta patología es de 44,5 años y generalmente afecta las glándulas salivares mayores. Presentamos aquí el caso clínico de una mujer de 25 años de edad que el año 2002 consultó por un aumento de volumen de la parótida derecha, de larga data. Después de estudios diagnósticos y tratamientos fue intervenida quirúrgicamente y el análisis histopatológico sugirió el diagnóstico APEP.

Sclerosing polycystic adenosis (SPA) of the parotid gland is a rare disease characterized histologically by a reactive inflammation of the salivary glands, although the presence of displasia and atypia raise the possibility that SPA might represent a neoplastic lesion. SPA bears histopathological resemblance to fibrocystic disease of the breast, and both glands show progesterone and oestrogen receptors in the ductal cells. The mean age of occurrence is 44.5 year-old, and it mostly affects major salivary glands. We report the case of a 25-years-old woman, who in 2002 presented with increased volume in the right parotid gland. After medical studies and several surgical treatments, the histopathological study revealed it to be SPA.

Proinflammatory cytokines tumor necrosis factor-alpha and interferon-gamma alter tight junction structure and function in the rat parotid gland Par-C10 cell line.

Sjögren's syndrome (SS) is an autoimmune disorder characterized by inflammation and dysfunction of salivary glands, resulting in impaired secretory function. The production of the proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) is elevated in exocrine glands of patients with SS, although little is known about the effects of these cytokines on salivary epithelial cell functions necessary for saliva secretion, including tight junction (TJ) integrity and the establishment of transepithelial ion gradients. The present study demonstrates that chronic exposure of polarized rat parotid gland (Par-C10) epithelial cell monolayers to TNF-alpha and IFN-gamma decreases transepithelial resistance (TER) and anion secretion, as measured by changes in short-circuit current (I(sc)) induced by carbachol, a muscarinic cholinergic receptor agonist, or UTP, a P2Y(2) nucleotide receptor agonist. In contrast, TNF-alpha and IFN-gamma had no effect on agonist-induced increases in the intracellular calcium concentration [Ca(2+)](i) in Par-C10 cells. Furthermore, treatment of Par-C10 cell monolayers with TNF-alpha and IFN-gamma increased paracellular permeability to normally impermeant proteins, altered cell and TJ morphology, and downregulated the expression of the TJ protein, claudin-1, but not other TJ proteins expressed in Par-C10 cells. The decreases in TER, agonist-induced transepithelial anion secretion, and claudin-1 expression caused by TNF-alpha, but not IFN-gamma, were reversible by incubation of Par-C10 cell monolayers with cytokine-free medium for 24 h, indicating that IFN-gamma causes irreversible inhibition of cellular activities associated with fluid secretion in salivary glands. Our results suggest that cytokine production is an important contributor to secretory dysfunction in SS by disrupting TJ integrity of salivary epithelium.

Anatomía microscópica de las glándulas salivales por medio de una técnica histológica convencional y no convencional / Microscopio anatomy of the salivary glands for mean of conventional and not-conventional histologic technique

Las técnicas de fijación y conservación permiten detener los procesos de desorganización de los tejidos y son necesarios para analizar la anatomía microscópica de ellos. El propósito de este estudio fue analizar las características histológicas de las glándulas parótida y submandibular obtenidas a partir de tres cadáveres humanos fijados y conservados mediante: a) solución conservadora en base a formaldehido (muestra I) y b) cámara de frío por 12 horas (muestra II), ambas muestras procesados para hematoxilina-eosina (H-E); c) plastinación con resina epóxica (muestra III) y procesado para H-E y con azul de metileno- eosina sin inclusión previa. Se analizaron las características de los adenómeros y sistema de conductos glandulares. Las mejores características se encontraron en la muestra II, con un buen nivel de detalle en el parénquima glandular, una mayor basofilia se presentó en la muestra I. La muestra III presentó un bajo nivel de detalle a la observación microscópica, los mejores resultados se obtuvieron utilizando azul de metileno. Las mayores dificultades en el procesamiento histológico de las piezas plastinadas se encontraron en el corte y en el tiempo necesario para la tinción. Los resultados sugieren que es posible obtener preparaciones histológicas a partir de necropsias en cadáveres fijados y conservados para la docencia e investigación anatómica.

The techniques of fixation and conservation allow to stop the processes of tissues disorganization and they are necessary to analyze the microscopic anatomy of them. The purpose of this study was to analyze the histologic characteristic of the parotid and submandibular glands obtained from three human cadavers fixed and conserved by means of: a) conservative solution based on formaldehyde (Sample I) and b) camera of cold for 12 hours (Sample II), both samples processed for hematoxilin-eosin (H-E); c) plastination with epoxic resin (Sample III) and processed for H-E and with methylene blue - eosin without previous inclusion. The characteristics of the adenomer and glandular ducts system were analyzed. The best characteristics were in the sample II, with a good detail level in the glandular parenchyma, a greater basophilia was presented in the sample I. The sample III it presented a low detail level to the microscopic observation, the best results were obtained using methylene blue. The biggest difficulties in the histologic process of the plastinated specimens were in the cut and time for tintion. The results suggest that it is possible to obtain histologic preparations from autopsies in fixed cadavers and conserved for teaching and anatomical investigation.