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Objective: This study assessed the effect of cevimeline and different concentrations of gum arabic on the parotid gland of rats being given xerostomia-inducing methotrexate. Methods: One hundred twenty-five rats were divided into five equal groups of twenty-five each. The rats in Group I received basic diets, while those in Groups II, III, IV, and V received 20 mg/kg MTX as a single intraperitoneal dose on day one. Group III received 10 mg/kg CVM dissolved in saline orally and daily, and the other two groups received a 10% W/V aqueous suspension of GA. Therefore, Group IV received 2 ml/kg suspension orally and daily, while Group V received 3 ml/kg suspension orally and daily. After 9 days, the parotid glands were dissected carefully and prepared for hematoxylin and eosin (H&E) staining as a routine histological stain and caspase-3 and Ki67 immunohistochemical staining. Quantitative data from α-Caspase-3 staining and Ki67 staining were statistically analysed using one-way ANOVA followed by Tukey's multiple comparisons post hoc test. Results: Regarding caspase-3 and Ki67 immunohistochemical staining, one-way ANOVA revealed a significant difference among the five groups. For Caspase-3, the highest mean value was for group II (54.21 ± 6.90), and the lowest mean value was for group I (15.75 ± 3.67). The other three groups had mean values of 31.09 ± 5.90, 30.76 ± 5.82, and 20.65 ± 3.47 for groups III, IV, and V, respectively. For Ki67, the highest mean value was for group I (61.70 ± 6.58), and the lowest value was for group II (18.14a ± 5.16). The other three groups had mean values of 34.4 ± 9.27, 48.03 ± 8.40, and 50.63 ± 8.27 for groups III, IV, and V, respectively. Conclusion: GA, rather than the normally used drug CVM, had a desirable effect on the salivary glands of patients with xerostomia. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the performance of diffusion-weighted magnetic resonance imaging (DW-MRI) as a noninvasive tool for assessing parotid gland function in type 2 diabetes mellitus (T2DM) patients. Methods: T2DM patients (n = 40, male, age 40–60 years) and healthy controls (n = 40) were examined. Salivary function was assessed using 1.5D echo-planar diffusion-weighted imaging. The examinations were performed before and every 3 min until 10 times after acid stimulation with a 500-mg vitamin C pill. The maximum apparent diffusion coefficient (ADC) in the parotid glands (pADCmax) and time to peak ADC in the parotid glands (pTmax) during stimulation were obtained. ADC values at every time point and the peak value of ADC were compared between the two groups and statistically analyzed. Results: The ADC values in the parotid gland in the resting state in the patient group were slightly lower than those in the healthy controls ((1.02 ± 0.08) × 10−3 mm2/s vs. (1.11 ± 0.09) × 10−3 mm2/s, p > 0.05). The ADCs in the healthy controls increased after stimulation until they gradually reached their peak. The ADCs in the patient group first decreased after stimulation and then gradually increased until they peaked. The pADCmax in the patient group was significantly lower than that in the healthy controls ((1.45 ± 0.08) × 10−3 mm2/s vs. (1.7 ± 0.06) × 10−3 mm2/s, p < 0.05). The peak ADCs in the parotid gland and relative signal intensity after acid stimulation were significantly correlated (r = 0.666, p < 0.05). Conclusion: DW-MRI before and after acid stimulation is potentially useful for noninvasive prediction of diabetes-induced xerostomia severity. ADC value is a sensitive indicator of parotid gland dysfunction and may indirectly reflect the degree of this dysfunction caused by fat deposition. [ABSTRACT FROM AUTHOR]

The study was designed to evaluate the influence of hyperbaric oxygenation therapy (HBOT) on the parotid gland in patients following radiotherapy for head and neck tumours. HBOT response was monitored by 3T magnetic resonance imaging (MRI) using T2 mapping and subsequent measurement of mean T2 and T2 variability as well as by salivary tests (salivary flow, buffer capacity, and pH). Eighteen patients previously treated with irradiation doses between 50 and 80 Gy as well as 18 healthy gender and age matched controls were enrolled. MRI was performed prior to HBOT (40.2 ± 20 months after radiotherapy) and after 20 daily HBOT at 2.5 ATA (absolute atmosphere). Each HBOT consisted of breathing 100% oxygen for 90 minutes. Significant differences in mean T2 prior to HBOT were observed between the ipsilateral irradiated (121 ± 20 ms), contralateral parotids (107 ± 21) and control group (96 ± 12 ms). A positive correlation in patients between T2 variability and irradiation dose was detected in contralateral parotids before HBOT (R = 0.489, p = 0.0287). In addition, negative correlations were observed between mean T2 in the ipsilateral as well as the contralateral gland and salivary flow before and after HBOT. Negative correlations between mean T2, T2 variability and pH of unstimulated saliva were also observed in the sides of parotid before and after HBOT. The study confirmed that T2 mapping had a potential for monitoring the differences between irradiated and normal parotid glands. It could also be useful in the assessment of the glandular tissue response to HBOT. [ABSTRACT FROM AUTHOR]

AIM: Mandibular condylar fractures account for 25 to 52 % of all mandibular fractures. Though current literature favors open reduction and internal fixation (ORIF) of condylar-base and low condylar-neck fractures, extraoral approaches are usually considered to be complicated by the risk of facial nerve injury and other possible complications. This study was undertaken to demonstrate that the periangular transmasseteric infraparotid surgical approach (TMIP) to condylar-base and low condylar-neck fractures provides excellent access to the bony fragments with minimal risk of complications such as facial nerve and parotid gland injury. PATIENTS: In the period from January 2010 to December 2018, 81patients (96 fractures) with condylar-base and low condylar-neck fractures underwent ORIF via periangular transmasseteric infraparotid surgical approach. RESULTS: The results of this retrospective study showed minimal postoperative complications. The periangular transmasseteric infraparotid surgical approach allowed precise anatomic repositioning and fixation of the bony fragments in almost all cases except for two juvenile cases with noticeable scars and one case with plate fracture. There were no transient or permanent facial nerve palsies, parotid gland or salivary fistulae complications during a 12-month follow-up period. CONCLUSION: The periangular infraparotid transmasseteric approach to ORIF of condylar-base and low condylar-neck fractures is an effective and safe approach allowing accurate anatomic reposition and fixation of the fragments with minimum surgical complications. [ABSTRACT FROM AUTHOR]

Myeloid sarcoma (MS) is a rarely encountered extramedullary localized tumor that is composed of immature myeloid cells. We reported an extremely rare case of MS with concurrent bone marrow (BM) involvement that invaded into a preexisting sebaceous lymphadenoma in the parotid gland and neck lymph nodes. Prompted by this case, we also present a literature review of MS invasion into salivary glands. A 62-year-old man was initially diagnosed with carcinoma that arose in a sebaceous lymphadenoma in the parotid gland, through a total parotidectomy with neck dissection. After an extensive histopathological review that included immunohistochemistry, a pathologic diagnosis of MS with infiltration into the sebaceous lymphadenoma with concurrent BM involvement was confirmed. MS is difficult to diagnose accurately; herein, we analyzed the clinical presentations and effectiveness of the various diagnostic methods with a review of the literature. 0ere are 17 cases, including our case, reported in 13 studies. Of the cases in which the salivary glands were affected, 10 involved the parotid gland, six involved the submandibular gland, and one involved both. Isolated invasion of the salivary gland was found in one case of parotid gland invasion and three cases of submandibular gland invasion. In 13 cases, the salivary glands were affected by various other lesions. Although there were no incidences of isolated MS, six patients were diagnosed with secondary MS and eight patients with MS with BM involvement, including this case. 0e diagnosis of MS is difficult given its rarity, and a high index of suspicion and integrated radiologic and careful histopathologic evaluation are required. Most cases of MS infiltrating the salivary gland might be indicated by the possibility of BM involvement. MS with BM involvement predicts poor prognosis and the need for intensive systemic treatment. [ABSTRACT FROM AUTHOR]

Juvenile recurrent parotitis (JRP) is a recurrent parotid inflammation of nonobstructive, nonsuppurative nature. It manifests in childhood and usually resolves after puberty but may also persist into adulthood. JRP is characterized by recurrent episodes of unilateral or/and bilateral parotid swelling with pain, reduction of salivary secretion, swallowing difficulty, fever, and malaise. The cause of this condition remains obscure. Throughout the last two decades, many therapeutic methods have been used in order to reduce the frequency and severity of JRP. During the acute episodes, conservative approaches (antibiotics, analgesics, sialogogues, massage of the parotid gland, and mouth rinses) are used. Parotidectomy has been suggested in rare selective occasions. Recently, a promising concept of sialendoscopy, which is a minimal invasive endoscopic technique, has been applied. This review outlines the literature on JRP focusing on methods and challenges in diagnosing JRP along with the differential diagnosis of JRP and the function of the parotid during JRP. In addition, we describe the treatment options for JRP, pointing out the importance of sialendoscopy as a diagnostic and treatment procedure that offers improvement in patients' daily life. [ABSTRACT FROM AUTHOR]

Skin secretions of frogs have a high chemical complexity. They have diverse types of biomolecules, such as proteins, peptides, biogenic amines, and alkaloids. These compounds protect amphibians' skin against growth of bacteria, fungi, and protozoa and participate in defense system against attack from predators. Therewith, this work performed biochemical and biological profile of macroglands parotoid secretion from cane toad. For poison analysis, we performed molecular exclusion and reverse phase chromatography, electrophoresis, and mass spectrometry. Antimicrobial, antiplasmodial, leishmanicidal, cytotoxicity, genotoxicity, and inflammatory activity of crude and/or fractions of R. marina secretion were also evaluated. Fractionation prior to filtration from poison showed separation of low mass content (steroids and alkaloids) and high molecular mass (protein). Material below 10 kDa two steroids, marinobufagin and desacetylcinobufagin, was detected. Crude extract and fractions were active against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Plasmodium falciparum, Leishmania guyanensis, and Leishmania braziliensis. Crude extract was also active against cancer cells although it was not cytotoxic for normal cells. This extract did not show significant DNA damage but it showed an important inflammatory effect in vivo. The information obtained in this work contributes to the understanding of the constituents of R. marina secretion as well as the bioactive potential of these molecules. [ABSTRACT FROM AUTHOR]

The parotid gland is drained by the parotid duct which normally measures 50 mm in length and 3 mm in width. The parotid duct emerges at the anterior border of the gland and runs horizontally across the masseter muscle to pierce the buccinator and open into the vestibule of the mouth. The occurrence of double parotid duct is 7% and there are only very few literature references. The parotid gland develops as an epithelial bud arising from the oral ectoderm and invaginating into the underlying mesenchyme. The proximal part canalises to form the duct and the distal end diferentiates to form the acini. An early division of the parotid duct with acini from both ducts intermingling with each other results in the formation of double parotid duct. The cytodiferentiation and morphogenesis of parotid gland are infuenced by cell specifc gene expression and cell-matrix interactions that produce collagen responsible for branching and hyaluronidase responsible for acini formation. The awareness of presence of a double parotid duct is signifcant in ductal endoscopic procedures and in performing surgery in the parotid region. [ABSTRACT FROM AUTHOR]

Introduction. Angiofollicular lymph node hyperplasia (Castleman’s disease) is a nonmalignant lymphoproliferative disorder that generally involves the lymph nodes of young adults, most commonly in the mediastinum. Rarely, Castleman’s disease may present in the parotid gland. The disease can be further classified into unicentric or multicentric forms, with considerable differences in presentation, treatment, and prognosis. Case(s). We present cases of two pediatric patients, aged 7 and 11, who both presented with a slow-growing, painless parotid mass. In each case, the mass was excised via a superficial parotidectomy and the diagnosis made postoperatively upon further pathologic examination. At 6 months of follow-up, both had fully intact facial nerve function and no evidence of recurrence. Discussion. Castleman’s disease presents a diagnostic challenge in the head and neck region, as radiographic characteristics and fine needle aspiration results are often inconclusive. Definitive diagnosis requires surgical excision for pathologic examination. The unicentric form generally presents as a painless mass and can be successfully treated with complete excision. The multicentric form is associated with constitutional symptoms and its treatment remains controversial. Conclusion. Although rare, clinicians should be aware of both forms of Castleman’s disease when creating a differential diagnosis for parotid masses. [ABSTRACT FROM AUTHOR]

Xerostomia is a common side effect of ionizing radiation used to treat head and neck cancer. A groundbreaking Phase I human clinical trial using Adenoviral gene transfer of Aquaporin-1 (AQP1) to a single salivary gland of individuals suffering from radiation-induced xerostomia has recently been reported. Unfortunately, the limitations of the Adenoviral vector system used in this pioneering trial preclude its advancement to a Phase II trial, and we have thus undertaken to evaluate the therapeutic potential of ultrasound-assisted nonviral gene transfer (UAGT) as an alternative means of delivering AQP1 gene therapy to the salivary gland by comparing head-to-head with the canonical Adenoviral vector in a swine model. Swine irradiated unilaterally with a 10-Gy electron beam targeted at the parotid gland suffered from significant, sustained hyposalivation that was bilateral, despite irradiation being confined to the targeted gland. Unilateral AQP1 gene therapy with UAGT resulted in bilateral restoration of stimulated salivary flow at 48 h and 1 week post treatment (1.62±0.48 ml and 1.87±0.45 ml) to preinjury levels (1.34±0.14 ml) in a manner comparable to Adenoviral delivery (2.32±0.6 ml and 1.33±0.97 ml). UAGT can replace the Adenoviral vector as a means of delivering AQP1 gene therapy in the irradiated swine model, and it is a candidate for advancement to a Phase I human clinical trial. [ABSTRACT FROM AUTHOR]

The aim of this prospective open-label study was to treat disabling drooling in children with cerebral palsy (CP) with onabotulinumtoxin A (A/Ona, Botox®) into submandibular and parotid glands and find the lowest effective dosage and least invasive method. A/Ona was injected in 14 children, Mean age 9 years, SD 3 years, under ultrasonic guidance in six successive Series, with at least six months between injections. Doses and gland involvement increased from Series A to F (units (U) per submandibular/parotid gland: A, 10/0; B, 15/0; C, 20/0; D, 20/20; E, 30/20; and F, 30/30). The effect was assessed 2, 4, 8, 12, and 20 weeks after A/Ona (drooling problems (VAS), impact (0-7), treatment effect (0-5), unstimulated whole saliva (UWS) flow and composition)) and analyzed by two-way ANOVA. The effect was unchanged-moderate in A to moderate-marked in F. Changes in all parameters were significant in E and F, but with swallowing problems ≤5 weeks in 3 of 28 treatments. F had largest VAS and UWS reduction (64% and 49%). We recommend: Start with dose D A/Ona (both submandibular and parotid glands and a total of 80 U) and increase to E and eventually F (total 120 U) without sufficient response. [ABSTRACT FROM AUTHOR]

The goal of this study was to evaluate the actual anatomical and dosimetric changes of parotid glands in nasopharyngeal carcinoma patients during intensity modulated radiation therapy. With helical tomotherapy, its planning system, and adaptive software, weekly anatomical and dosimetric changes of parotid glands in 35 NPC patients were evaluated. Interweekly parotid volume varied significantly (P<0.03). The rate of volume change reached the highest level at the 16th fraction. The average V1 increased by 32.2 (left) and 28.6 (right), and the average D50 increased by 33.9 (left) and 24.93 (right), respectively. Repeat data comparison indicated that the V1 and D50 varied significantly among different fractions (both with P=0.000). The variation of parotid volume was inversely correlated with that of the V1 and D50 (both with P=0.000). In conclusion, parotid volume and actual dose vary significantly in NPC patients during IMRT. Replanning at the end of the fourth week of IMRT may have clinical benefits. [ABSTRACT FROM AUTHOR]

Secretory proteins produced by salivary glands are stored in granules and released into saliva. Rodent salivary glands are a reliable experimental model because they are morphologically and functionally similar to those of humans. To determine if the effects of microgravity on secretory proteins are increased on extended flights, their expression in mouse parotid glands, morphological, immunocytochemical, and biochemical/molecular methods were employed. Acinar cells of STS-135 (13 day) and Bion-M1 (30 day) flight animals showed an increase of autophagy and apoptosis, while duct cells contained vacuoles with endocytosed proteins. In STS-135, decreases were seen in the regulatory subunit of type II protein kinase A (RII) by Western blotting, and demilune cell and parotid protein (DCPP) and α- amylase (p<0.01) by immunogold labeling, while proline-rich proteins (PRPs, p<0.001) and parotid secretory protein (PSP, p<0.05) were increased. These results suggest microgravity effects on secretion are function-dependent. Microarray analyses showed significant changes in the expression of a number of genes, including components of the cyclic-3',5',-adenosine monophosphate (cyclic AMP) signaling pathway. Compared to habitat ground controls, mice from both flights exhibited altered expression of cyclic AMP-specific phosphodiesterases, adenylate cyclase isoforms, and several A-kinase anchoring proteins. Bion-M1 flight mice showed increases in gene expression for lysozyme and amylase, a decrease in PRPs, and RII expression was unchanged from control values. Secretory protein expression is altered by travel in space, representing a reversible adjustment to microgravity conditions. Ultimately, the goal is to develop a test kit using saliva -- an easily obtained body fluid -- to assess the physiologic effects of travel in space. [ABSTRACT FROM AUTHOR]

Background To determine the position and dose delivery changes rules of parotid gland (PG) during the course of intensity modulated radiation therapy (IMRT) in nasopharyngeal carcinoma patients (NPC). Materials and methods One hundred and forty one competed tomography (CT) images from 47 NPC patients (three images for each patient were acquired before treatment, at the 15th and 25th fraction during the treatment) who underwent radical IMRT were selected for this study. A total of 70-76Gy at 2.12-2.3 Gy/fraction/d was given to the GTVnx in 33 fractions. The distances between the lateral/medial aspects of PG and midline (DLM and DMM) at the level of odontoid process were measured. The dose differences between plan and actual delivery were calculated. Results The volume reductions of PGs between fractions 15 and 25 were larger than those in the first 15 fractions (4.68 ± 3.23 cc vs. 2.46 ± 4.55 cc for the right PG and 5.96 ± 2.99 cc vs. 2.06 ± 2.99 cc for the left PG). However, the percentage of gland volume receiving ≥30Gy (V30) of bilateral PGs decreased more significantly in the first 15 fractions than that between fractions 15 and 25 (5.61 ± 16.04% vs. 1.14 ± 21.54% for the right PG and 6.87 ± 15.58% vs. 0.81 ± 15.94% for the left PG). The gross tumor volume of the nasopharynx (GTVnx) decreased more significantly in the first 15 fractions than that between the 15th and 25th fraction (8.23 ± 13.61 cc vs. 3.30 ± 8.09 cc). The DMM of ipsilateral PGs reduced in the first 15 fractions (0.80 ± 2.96 mm) but increased between fraction 15 and 25 (-2.19 ± 3.96 mm). While ipsilateral PG shifted into target volume but shifted out target volume between fraction 15 and 25. Parotid glands V30 was correlated with GTVnx, GTVnx reduction and DMM reduction (p < 0.01). Conclusion Our results indicate that the reduction of GTVnx leads to the positional change of the parotid gland, which results in more significant dose change of the parotid gland in the first 15 fractions than that between fraction 15 and 25. [ABSTRACT FROM AUTHOR]

Basal cell adenoma is an uncommon type of monomorphous adenoma of the salivary gland. Its most frequent location is the parotid gland. Its recurrence rate and conversion to basal cell adenocarcinoma after inadequate surgical excision is high. We present a 68-year-old female patient with a basal cell adenoma of the upper lip arising from minor salivary gland. [ABSTRACT FROM AUTHOR]

Phospholipase D (PLD) has been implicated in many cellular functions, such as vesicle trafficking, exocytosis, differentiation, and proliferation. The aim of this study was to characterize the role of PLD in HSY cells, a human cell line originating from the intercalated duct of the parotid gland. As the function and intracellular localization of PLD varies according to cell type, initially, the intracellular localization of PLD1 and PLD2 was determined. By immunofluorescence, PLD1 and PLD2 both showed a punctate cytoplasmic distribution with extensive co-localization with TGN-46. PLD1 was also found in the nucleus, while PLD2 was associated with the plasma membrane. Treatment of cells with the primary alcohol 1-butanol inhibits the hydrolysis of phosphatidylcoline by PLD thereby suppressing phosphatidic acid (PA) production. In untreated HSY cells, there was only a slight co-localization of PLD with the clathrin coated vesicles. When HSY cells were incubated with 1-butanol the total number of clathrin coated vesicles increased, especially in the juxtanuclear region and the co-localization of PLD with the clathrin coated vesicles was augmented. Transmission electron microscopy confirmed that the number of Golgi-associated coated vesicles was greater. Treatment with 1-butanol also affected the Golgi apparatus, increasing the volume of the Golgi saccules. The decrease in PA levels after treatment with 1-butanol likewise resulted in an accumulation of enlarged lysosomes in the perinuclear region. Therefore, in HSY cells PLD appears to be involved in the formation of Golgi associated clathrin coated vesicles as well as in the structural maintenance of the Golgi apparatus. [ABSTRACT FROM AUTHOR]

Aim: In recent years, tularemia's importance has been reemerging in Turkey. However, the majority of tularemia cases were reported in the northern cities of Turkey. In this study, we report 16 oropharyngeal tularemia patients living in rural areas of the city of Konya, which is located in the southern zone of the Central Anatolia Region of Turkey. Materials and methods: Sixteen patients (12 males, 4 females; mean age: 33.7 years, range: 10-73 years) were included in this study. All patients were admitted to our clinic for the evaluation of intractable cervical mass lesions. The diagnosis of tularemia was made after exclusion of other possible causes of the neck masses and the presence of a positive tularemia agglutination test. Each patient was treated with streptomycin and doxycycline for at least 3 weeks. Results: All patients suffered oropharyngeal stiffness. There were neck mass and tonsillitis on the same side in 2 patients, neck mass and acute pharyngitis in 3 patients, and only swelling in the neck region in 11 patients. One of the patients with tonsillitis had common maculopapular skin rashes with fever. Thirteen patients had mass at level 2 zones, 11 patients had mass on the right side of the neck, and 5 patients had mass on the left side of the neck. The right parotid gland was involved in 1 patient. Nine patients were successfully treated with only antimicrobial agents. Lymph node excision was performed in 6 patients, and parotidectomy was performed in 1 patient. Conclusion: This is the second series of tularemia from the city of Konya in Turkey. Tularemia should be considered in the differential diagnosis of patients with intractable neck mass, because early diagnosis and treatment may prevent surgical interventions. [ABSTRACT FROM AUTHOR]

Objective. The relationship of cryoglobulinaemia with lymphoproliferation of mucosa-associated lymphoid tissue (MALT) as risk factors for lymphoma evolution in SS remains to be clarified. The different biologic background of SS-related cryoglobulinaemia as compared with cryoglobulinaemia linked to HCV infection was clarified by different clinical and biologic approaches.Methods. B-cell clonal expansion was analysed in the bone marrow of 27 consecutive cases with primary SS and mixed cryoglobulinaemia, HCV unrelated, in comparison with 55 HCV-related patients with cryoglobulinaemic vasculitis (CV) without SS. The results were related to the possible occurrence and localization of B-cell lymphoma in the single case. Secondly, the prevalence of mixed cryoglobulinaemia was investigated in 41 unselected patients with primary SS showing either parotid myoepithelial sialadenitis (MESA) or a frank B-cell non-Hodgkin's lymphoma. Thirdly, the levels of serum cryoglobulins and RF were followed in one patient with primary SS, CV and parotid B-cell lymphoma of MALT after bilateral subtotal parotidectomy.Results. A polyclonal pattern of B expansion in the bone marrow was significantly more frequent in SS-related (19/27 cases) than in HCV-related cryoglobulinaemia (19/55) (P = 0.003). Cryoglobulins were positive in a fraction of patients with SS and malignant lymphoma or with parotid MESA (13/18 and 7/23, respectively), whereas MALT involvement by the lymphoproliferative disorder was the rule. Finally, the levels of serum cryoglobulins and RF markedly decreased in the SS patient undergoing bilateral subtotal parotidectomy.Conclusion. Lymphoproliferation of MALT appears as the biologic background of cryoglobulinaemia in SS, differently from HCV-related cryoglobulinaemia. [ABSTRACT FROM PUBLISHER]

Development of facial nerve palsy (FNP) may lead to dramatic change in the patient's facial function, expression, and emotions. The ophthalmologist may play an important role in the initial evaluation, and the long-term management of patients with new-onset of FNP. In patients with expected temporary facial weakness, no efforts should be wasted to ensure proper corneal protection. Patients with permanent functional deficit may require combination of surgical procedures tailored to the patient's clinical findings that may require good eye comfort and cosmesis. [ABSTRACT FROM AUTHOR]

Presents information on a study which investigated the relationship between sodium/hydrogen exchanger isoform 1 and the response of parotid glands to beta-adrenergic agonists. Methodology; Results of the study; Discussion.

Oral dryness causes significant health problems both functional (difficulty speaking, chewing and swallowing) and structural in teeth (increased number of infections) and oral mucosa. The main objective of this study is to show an alternative treatment to help stimulate the salivary secretion thus improving the quality of life of the patient. In this study, a salivary stimulation equipment using vibrotactile stimuli is shown. The system has been placed bilaterally in the parotid glands and assessed the efficacy of the salivary secretion by sialometry before and after the stimulation. The new proposal is capable of stimulating salivary secretion, in a significative way after 7 minutes of use, at least in the cases analyzed, and fulfills low-cost, easy-to-use, and safe technical restrictions. In this setting, this paper suggests the performance of a deep clinical trial to measure the exact efficacy of the prototype and the times and frequencies needed to state the optimal treatment depending in each case., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2022 Francisco Gómez-García et al.)

Presents information on a study which examined the influence of age on the variability of parotid salivary gland flow rates. Methodology; Results of the study; Discussion.

Focuses on a study which described experiments indicating the suitability of one of the parotid gland cell lines, Par-C10, in Ussing chamber studies. Methodology used on cell culture, morphological evaluation and measurement of short-circuit current in Par-C10 monolayers; Elaboration on the results; Conclusion.

Focuses on the capability of mouse parotid acinar cells to synthesize IL-1beta and IL-6 and to release the cytokines from their granules after alpha- and beta-adrenergic stimulation. In situ hybridization of IL-1beta and IL-6 mRNA in parotid gland; Release of IL-1beta, IL-6, and amylase from parotid acinar cells; Ultrastructure localization of IL-1beta and IL-6.

If optimal investment in anti-predator defences depends on predation risk, invading new regions (and thus, encountering different predators) may favour shifts in that investment. Cane toads offer an ideal system to test this prediction: expensive anti-predator toxins are stored mainly in parotoid glands whose dimensions are easy to measure, and toad invasions have changed the suites of predators they encounter. Although plasticity may influence parotoid morphology, comparisons between parents and progeny revealed that gland dimensions were highly heritable. That heritability supports the plausibility of an evolved basis to variation in gland dimensions. Measurements of 3779 adult toads show that females have larger glands than males, invasive populations have larger glands than in the native-range, and that parotoid sexual size dimorphism varies strongly among invaded areas. Geographic variation in parotoid morphology may be driven by predation risk to both adult toads and offspring (provisioned with toxins by their mother), with toxins allocated to eggs exacerbating the risk of cannibalism but reducing the risk of interspecific predation. Investment into chemical defences has evolved rapidly during the cane toad's international diaspora, consistent with the hypothesis that organisms flexibly adjust resource allocation to anti-predator tactics in response to novel challenges.

Previous studies revealed high incidence of acquired N-glycosylation sites acquired N-glycosylation sites in RNA transcripts encoding immunoglobulin heavy variable region (IGHV) 3 genes from parotid glands of primary Sjögren's syndrome (pSS) patients. In this study, next generation sequencing was used to study the extent of ac-Nglycs among clonally expanded cells from all IGVH families in the salivary glands of pSS patients. RNA was isolated from parotid gland biopsies of five pSS patients and five non-pSS sicca controls. IGHV sequences covering all functional IGHV genes were amplified, sequenced, and analyzed. Each biopsy recovered 1,800-4,000 unique IGHV sequences. No difference in IGHV gene usage was observed between pSS and non-pSS sequences. Clonally related sequences with more than 0.3% of the total number of sequences per patient were referred to as dominant clone. Overall, 70 dominant clones were found in pSS biopsies, compared to 15 in non-pSS. No difference in percentage mutation in dominant clone-derived IGHV sequences was seen between pSS and non-pSS. In pSS, no evidence for antigen-driven selection in dominant clones was found. We observed a significantly higher amount of ac-Nglycs among pSS dominant clone-derived sequences compared to non-pSS. Ac-Nglycs were, however, not restricted to dominant clones or IGHV gene. Most ac-Nglycs were detected in the framework 3 region. No stereotypic rheumatoid factor rearrangements were found in dominant clones. Lineage tree analysis showed in four pSS patients, but not in non-pSS, the presence of the germline sequence from a dominant clone. Presence of germline sequence and mutated IGHV sequences in the same dominant clone provide evidence that this clone originated from a naïve B-cell recruited into the parotid gland to expand and differentiate locally into plasma cells. The increased presence of ac-Nglycs in IGHV sequences, due to somatic hypermutation, might provide B-cells an escape mechanism to survive during immune response. We speculate that glycosylation of the B-cell receptor makes the cell sensitive to environmental lectin signals to contribute to aberrant B-cell selection in pSS parotid glands.

Objective: To evaluate the early changes in the apparent diffusion coefficient (ADC) of the salivary glands during radiotherapy (RT) and their association with the degree of xerostomia at 6 months after RT in patients with nasopharyngeal carcinoma (NPC)., Materials and Methods: We enrolled 26 patients with NPC who underwent RT. Each patient underwent diffusion-weighted MRI of the salivary glands at rest and with gustatory stimulation within 1 week before RT and 2 weeks after the beginning of RT. The ADC at rest (ADC R ) and increase and increase rate with stimulation (ADC I , ADC IR ) of the submandibular and parotid glands were calculated. The differences in the variables' values between 2 weeks after the beginning of RT and baseline (ΔADC R , ΔADC I , and ΔADC IR ) were compared to the degree of xerostomia at 6 months after RT., Results: The ADC R of the submandibular and parotid glands were both significantly higher at 2 weeks after the beginning of RT than found at baseline (both p < 0.01). The ADC I and ADC IR for the parotid glands were both significantly lower at 2 weeks after the beginning of RT than found at baseline (both p < 0.01). ΔADC I and ΔADC IR of the parotid glands were associated with the degree of xerostomia at 6 months after RT ( r = -0.61 and -0.72, both p < 0.01)., Conclusion: The ADCs of the salivary glands change early during RT. The differences in the ADC increase and increase rate of the parotid glands between 2 weeks after the beginning of RT and baseline were associated with the degree of xerostomia at 6 months after RT.

Background: Shear wave elastography is a relatively new method of quantitative measurement of tissue elasticity. Assuming that malignant lesions are stiffer than benign ones, elastography may provide supplementary information for their discrimination. However, potential confounding factors impacting tissue stiffness should be investigated first., Aims: The objective of this study was to measure the stiffness of selected tissues of the head and neck in a normal population and to evaluate its relationship to age, sex, and body mass index., Methods: Stiffness of the thyroid, submandibular and parotid glands, masseter and sternocleidomastoid muscles, and cervical lymph nodes was measured bilaterally in 128 healthy volunteers (83 female and 45 male). At least 20 subjects in each decade of life (20-29, 30-39‥, 70+) were enrolled. Shear wave elastography was performed by a single radiologist in all the subjects. The stiffnesses obtained were correlated with age, sex, and body mass index., Results: The mean stiffness was 9.5 ± 3.6 kPa for the thyroid, 9.5 ± 4.6 kPa for the lymph node, 11.0 ± 3.4 kPa for the submandibular gland, 9.0 ± 3.5 kPa for the parotid gland, 9.9 ± 4.1 kPa for the sternocleidomastoid, and 10.0 ± 4.3 kPa for the masseter muscle. A slight general decrease in stiffness with increasing age was found. BMI and weight had a small impact on the minimum and maximum stiffness values. The sex of the subject did not affect elasticity., Conclusion: The mean stiffness of healthy head and neck organs has a relatively narrow distribution around 11 kPa. The changes of stiffness with age, BMI, and weight that were identified are too small to have clinical impact.

Presents an abstract of the paper entitled `Volume-sensitive Chloride Channels in Rat Parotid Acinar Cells,' by Jorge Arreola, James E. Melvin, and Ted Begenisich presented at the 1994 meeting of the Society of General Physiologists at the Marine Biological Laboratory in Woods Hole, Massachusetts. Characterization of the volume-sensitive chloride channels in parotid acinar cells.

Background: Saliva is a complex fluid, whose important role is to maintain the well being of oral cavity. Salivary gland hypofunction or hyposalivation is the condition of having reduced saliva production which leads to the subjective complaint of oral dryness termed xerostomia.(7) Management of xerostomia includes palliative therapy using topical agents or systemic therapy. Electrostimulation to produce saliva was studied in the past and showed moderate promise but never became part of mainstream therapy. Hence, this study was undertaken to evaluate the effect of transcutaneous electrical nerve stimulation (TENS) on whole salivary flow rate in healthy adults and to evaluate how long this effect of TENS lasts on salivary flow., Materials and Methods: One hundred healthy adult subjects were divided into five age groups with each group containing 20 subjects equally divided into males and females in each group. Unstimulated saliva was collected using a graduated test tube fitted with funnel and quantity was measured. Transcutaneous electrical nerve stimulation unit was activated and stimulated saliva was collected. Saliva was again collected 30 minutes and 24 hours post stimulation., Results: The mean unstimulated whole saliva flow rate for all subjects (n = 100) was 2.60 ml/5 min. During stimulation, it increased to 3.60 ± 0.39 ml/5 min. There was 38.46% increase in salivary flow. Ninety six out of 100 responded positively to TENS therapy. Salivary flow remained increased 30 minutes and 24 hours post stimulation with the values being 3.23 ± 0.41 ml/5 min and 2.69 ± 0.39 ml/5 min respectively. Repeated measures One way analysis of variance (ANOVA) test showed that the difference between these values were statistically significant., Conclusion: Transcutaneous electrical nerve stimulation therapy was effective for stimulation of whole saliva in normal, healthy subjects and its effect retained till 30 minutes and a little up to 24 hours. Transcutaneous electrical nerve stimulation may work best synergistically with other sialagogues and can be used for the management of xerostomia.

The standard of care for head and neck cancer typically includes surgical resection of the tumor followed by targeted head and neck radiation. However depending on tumor location and stage, some cases may not require surgical resection while others may be treated with chemoradiation. Unfortunately, these radiation treatments cause chronic negative side effects for patients. These side effects are associated with damage to surrounding normal salivary gland tissue and include xerostomia, changes in taste and malnutrition. The underlying mechanisms of chronic radiation-induced salivary gland dysfunction are unknown, however, in rodent models persistently elevated proliferation is correlated with reduced stimulated salivary flow. The rapalogue, CCI-779, has been used in other cell systems to induce autophagy and reduce proliferation, therefore the aim of this study was to determine if CCI-779 could be utilized to ameliorate chronic radiation-induced salivary gland dysfunction. Four to six week old Atg5f/f; Aqp5-Cre, Atg5+/+; Aqp5-Cre and FVB mice were treated with targeted head and neck radiation. FVB mice were treated with CCI-779, chloroquine, or DMSO post-radiation. Stimulated salivary flow rates were determined and parotid and submandibular salivary gland tissues were collected for analyses. Mice with a defect in autophagy, via a conditional knockout of Atg5 in the salivary glands, display increased compensatory proliferation in the acinar cell compartment and hypertrophy at 24-72 hours following radiation. FVB mice treated with post-therapy CCI-779 have significant improvements in salivary gland physiology as determined by stimulated salivary flow rates, proliferation indices and amylase production and secretion. Consequently, post-radiation use of CCI-779 allows for improvement of salivary gland function and reestablishment of glandular homeostasis. As CCI-779 is already FDA approved for other uses, it could have a secondary use to alleviate the chronic side effects in head and neck cancer patients who have completed anti-tumor therapy.

The exocytosis of salivary proteins is mainly regulated by cAMP, although soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), which mediate cAMP-dependent exocytic membrane fusion, have remained unidentified. Here we examined the effect of isoproterenol (ISO) and cytochalasin D (CyD) on the level of SNARE complexes in rat parotid glands. When SNARE complexes were immunoprecipitated by anti-SNAP23, the coprecipitation of VAMP2 was significantly increased in response to ISO and/or CyD, although the coprecipitation of VAMP8 or syntaxin 4 was scarcely augmented. These results suggest that the SNAP23-VAMP2 interaction plays a key role in cAMP-mediated exocytosis from parotid glands., (Copyright © 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)

Ruminant nutrition relies upon the symbiotic relationship that exists with microbial populations in the rumen. Urea transported across the ruminal epithelia and secreted by the salivary glands is a key source of nitrogen for microbial growth in the rumen. As ruminal urea transport can be mediated by specific UT-B urea transporters, this study investigated whether UT-B urea transporters were also present in the bovine salivary gland. Western blotting experiments detected only small amounts of UT-B protein in whole-cell lysate from the bovine parotid gland. In contrast, strong 32 to 34 and 40 kDa UT-B proteins were detected in parotid plasma membrane-enriched protein, showing the importance of using enriched samples. These signals were also detected in rumen and correspond to bovine UT-B1 and UT-B2 urea transporters, respectively. Further immunolocalization studies identified that these proteins were located in the ductal system of the parotid gland. This study, therefore, confirmed the presence of UT-B urea transporter protein in the bovine parotid salivary gland., (Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The adult human parotid duct is roughly 6-8 cm long. From the parotid gland, parotid duct traverses through masseter muscle, penetrates through buccinator muscle, and opens into the oral cavity. This unique form of the parotid duct is likely correlated with the function of the duct, directly affected by the movement of the buccinator muscle during mastication and swallowing. Histological structure of the duct is known to be different in each region, and details of smooth muscle present in the parotid duct are mostly unclear. In this study, we conducted SEM and histological observations of the area where the parotid duct penetrates the buccinator muscle, and an observation of smooth muscle to investigate its existence using alpha-smooth muscle antibody. We confirmed the presence of an abundance of skeletal muscle bundles likely originating from the buccinator muscle under the epithelium of the parotid duct wall in the region where it penetrates the buccinator muscle. We also observed that some of the muscle fibers were completely attached to the epithelium. We observed a lack of smooth muscle in this region of the duct wall. From these results, we suggest that the area of the duct penetrating buccinator muscle plays a role in regulating the salivary passage through the contraction of the surrounding buccinator muscle fibers.

There is emerging consensus that P2X₄ and P2X₇ ionotropic purinoceptors (P2X₄R and P2X₇R) are critical players in regulating [Ca²⁺]i dynamics and fluid secretion in the salivary gland. In contrast, details regarding their compartmentalization and selective activation, contributions to the spatiotemporal properties of intracellular signals and roles in regulating protein exocytosis and ion channel activity have remained largely undefined. To address these concerns, we profiled mouse parotid acinar cells using live-cell imaging to follow the spatial and temporal features of ATP-evoked Ca²⁺ dynamics and exocytotic activity. Selective activation of P2X7Rs revealed an apical-to-basal [Ca²⁺]i signal that initiated at the sub-luminal border and propagated with a wave speed estimated at 17.3 ± 4.3 μm s⁻¹ (n =6). The evoked Ca²⁺ spike consisted of Ca²⁺ influx and Ca²⁺-induced Ca²⁺ release from intracellular Ca²⁺ channels. In contrast, selective activation of P2X₄Rs induced a Ca²⁺ signal that initiated basally and propagated toward the lumen with a wave speed of 4.3 ± 0.2 μm s⁻¹ (n =8) that was largely independent of intracellular Ca²⁺ channel blockade. Consistent with these observations, P2X₇R expression was enriched in the sub-luminal regions of acinar cells while P2X₄R appeared localized to basal areas. In addition, we showed that P2X₄R and P2X₇R activation evokes exocytosis in parotid acinar cells. Our studies also demonstrate that the P2X₄R-mediated [Ca²⁺]i rise and subsequent protein exocytosis was enhanced by ivermectin (IVR). Thus, in addition to furthering our understanding of salivary gland physiology, this study identifies P2X₄R as a potential target for treatment of salivary hypofunction diseases.

It has been shown that the voltage (V(m)) dependence of ClC Cl(-) channels is conferred by interaction of the protopore gate with H(+) ions. However, in this paper we present evidence which indicates that permeant Cl(-) ions contribute to V(m)-dependent gating of the broadly distributed ClC-2 Cl() channel. The apparent open probability (P(A)) of ClC-2 was enhanced either by changing the [Cl(-)](i) from 10 to 200 mM or by keeping the [Cl(-)](i) low (10 mM) and then raising [Cl(-)](o) from 10 to 140 mM. Additionally, these changes in [Cl(-)] slowed down channel closing at positive V(m) suggesting that high [Cl(-)] increased pore occupancy thus hindering closing of the protopore gate. The identity of the permeant anion was also important since the P(A)(V(m)) curves were nearly identical with Cl(-) or Br(-) but shifted to negative voltages in the presence of SCN() ions. In addition, gating, closing rate and reversal potential displayed anomalous mole fraction behaviour in a SCN(-)/Cl() mixture in agreement with the idea that pore occupancy by different permeant anions modifies the V(m) dependence ClC-2 gating. Based on the ec1-ClC anion pathway, we hypothesized that opening of the protopore gate is facilitated when Cl(-) ions dwell in the central binding site. In contrast, when Cl(-) ions dwell in the external binding site they prevent the gate from closing. Finally, this Cl(-)-dependent gating in ClC-2 channels is of physiological relevance since an increase in [Cl(-)](o) enhances channel opening when the [Cl(-)](i) is in the physiological range.

Salivary glands produce a HCO(3)(-)-rich fluid that is important for the neutral milieu in the upper gastrointestinal tract. The molecular mechanism of this secretion is poorly understood. Par-C10, an immortalized rat parotid acinar line, has been shown to secrete Cl(-)- in response to Ca(2+-)-mobilizing stimuli. Our aim was to assess the capacity of polarized monolayers of Par-C10 cells to transport and secrete HCO(3)(-)-. Transepithelial electrolyte movement was evaluated by short-circuit current measurements. Intracellular pH (pH(i)) was measured by microfluorometry in cells loaded with BCECF. Monolayers of Par-C10 cells, grown on Transwell membranes, developed high transepithelial resistance and exhibited vectorial anion secretion which was activated by both ATP and forskolin. The currents were partially inhibited by bumetanide and by withdrawal of HCO(3)(-) indicating the dependence of ion movements on NKCC and on HCO(3)(-) ions, respectively. In HCO(3)(-)-free solutions the recovery of pH(i) from acid loading was abolished by EIPA. In the presence of HCO(3)(-) there was a strong EIPA-insensitive recovery from acid loading which was inhibited by H(2)DIDS. ATP and forskolin stimulated HCO(3)(-) efflux from the cells. Furthermore, HCl(-) withdrawal experiments showed the presence of DNDS-sensitive basolateral anion exchange. In conclusion Par-C10 cells achieve transepithelial transport that is sensitive to both intracellular Ca(2+)- and cAMP-dependent stimulation. We identified Na(+)/H(+) exchange, Na(+)-HCO(3)(-) cotransport and anion exchange at the basolateral side of the cells as being involved in intracellular pH regulation and vectorial HCO(3)(-) secretion. This cell line offers a good model for further studies to understand the molecular mechanisms of salivary HCO(3)(-) secretion.

Mouse parotid acinar cells express P2X4 and P2X7 receptors (mP2X4R and mP2X7R) whose physiological function remains undetermined. Here we show that mP2X4R expressed in HEK-293 cells do not allow the passage of tetraethylammonium (TEA+) and promote little, if any, ethidium bromide (EtBr) uptake when stimulated with ATP or BzATP. In contrast, mP2X7R generates slowly decaying TEA+ current, sustained Na+ current and promotes robust EtBr uptake. However, ATP-activated TEA+ current from acinar cells was unlike that generated by mP2X7R or mP2X4R. Functional interactions between mP2X4R and mP2X7R were investigated in HEK cells co-transfected with different mP2X4 : mP2X7 cDNA ratios and using solutions containing either TEA+ or Na+ ions. Co-expressed channels generated a TEA+ current that displayed faster decay during ATP stimulation than mP2X7R alone. Moreover, cells transfected with a 2 : 1 cDNA ratio displayed decaying kinetics similar to those observed in acinar cells. Concentration-response curves in Na+-containing solutions were constructed for heterologously expressed mP2X4R, mP2X7R and mP2X4R:mP2X7R co-expressions as well as acinar cells. The EC50 values determined were 11, 220, 434 and 442 microM, respectively. Na+ currents generated by expressing mP2X4R or mP2X7R alone were potentiated by ivermectin (IVM). In contrast, IVM potentiation in acinar cells and HEK cells co-expressing P2X4 and P2X7 (1 : 1 or 2 : 1 cDNA ratios) was seen only when the ATP concentration was lowered from 5 to 0.03 mM. Taken together our observations indicate a functional interaction between murine P2X7 and P2X4 receptors. Such interaction might occur in acinar cells to shape the response to extracellular ATP in salivary epithelia.

Objective: To identify key target genes and activated signaling pathways associated with the pathogenesis of Sjögren's syndrome (SS) by conducting a systems analysis of parotid glands manifesting primary SS or primary SS/mucosa-associated lymphoid tissue (MALT) lymphoma phenotypes., Methods: A systems biology approach was used to analyze parotid gland tissue samples obtained from patients with primary SS, patients with primary SS/MALT lymphoma, and subjects without primary SS (non-primary SS controls). The tissue samples were assessed concurrently by gene-expression microarray profiling and proteomics analysis, followed by weighted gene-coexpression network analysis., Results: Gene-coexpression modules related to primary SS and primary SS/MALT lymphoma were significantly enriched with genes known to be involved in the immune/defense response, apoptosis, cell signaling, gene regulation, and oxidative stress. Detailed functional pathway analyses indicated that primary SS-associated modules were enriched with genes involved in proteasome degradation, apoptosis, signal peptides of the class I major histocompatibility complex (MHC), complement activation, cell growth and death, and integrin-mediated cell adhesion, while primary SS/MALT lymphoma-associated modules were enriched with genes involved in translation, ribosome biogenesis and assembly, proteasome degradation, class I MHC signal peptides, the G13 signaling pathway, complement activation, and integrin-mediated cell adhesion. Combined analyses of gene expression and proteomics data implicated 6 highly connected "hub" genes for distinguishing primary SS from non-primary SS, and 8 hub genes for distinguishing primary SS/MALT lymphoma from primary SS., Conclusion: Systems biology analyses of the parotid glands from patients with primary SS and those with primary SS/MALT lymphoma revealed pathways and molecular targets associated with disease pathogenesis. The identified gene modules/pathways provide further insights into the molecular mechanisms of primary SS and primary SS/MALT lymphoma. The identified disease-hub genes represent promising targets for therapeutic intervention, diagnosis, and prognosis.

Parotid glands secrete about 25% of all saliva produced in the salivary glands. In the presence of a stimulus, the amount of saliva secreted from the parotid gland increases to 50% (1). In human adults, the parotid duct, approximately 6-8 cm long, traverses the masseter muscle and penetrates through the buccinator muscle into the oral cavity. Although various studies have been conducted on the parotid gland, there are only few suggesting the functional roles of the parotid duct, especially of the area penetrating the buccinator muscle. In the present study, we observed parotid ducts of human fetuses to morphologically analyze the function of the buccinator muscles in the flux of parotid saliva. Thirty fetal specimens ranging from five to ten months of age were dissected for anatomical and histological examinations. The area of the parotid duct penetrating the buccinator muscle was fully formed in six-month-old fetuses. Furthermore, this study confirms the existence of thin buccinator muscle fibers underneath the epithelium of the parotid duct's distal portion. Results suggest that the buccinator muscle may play a major role in preventing the reflux of salivary secretions by assisting the contraction of the parotid duct.

Salivary gland acinar cells secrete large amounts of water and electrolytes, where aquaporins (AQPs) are thought to be involved in the secretion. In the present study, we investigated expression/localization of AQP6, and the anion transporting properties of AQP6 in mouse parotid acinar cells. RT-PCR, western blotting and immunohistochemical analyses revealed expression of AQP6 in acinar cells, localized in apical membrane. Voltage ramp from -100 mV to +100 mV at a holding potential of -60 mV elicited outwardly-rectifying currents, in the presence of extracellular Cl(-) channel blockers and intracellular solution with 150 mM Cs(+). These outward currents were increased when extracellular Cl(-) was replaced by Br(-), NO(3)(-), I(-), or SCN(-), accompanying a negative shift of reversal potentials. The outward current was enhanced by extracellular Hg(2+). These results were consistent with the biophysical properties of transfected AQP6 oocytes or HEK cells, which indicate that the AQP6 channel is functionally expressed in parotid acinar cells, and suggest that AQP6 contributes to secretion of anions in parotid acinar cells.

Background and Purpose: Our aim was to compare different field strengths monitoring physiologic changes due to oral stimulation of parotid glands by using diffusion-weighted (DW) echo-planar imaging (EPI)., Materials and Methods: Twenty-seven healthy volunteers were examined with a DW-EPI sequence at 1.5T and 3T before and after oral stimulation with commercially available lemon juice. The b factors used were 0, 500, and 1000 s/mm(2). Apparent diffusion coefficient (ADC) maps were evaluated with a manually placed region of interest including the entire parotid gland. For comparison of results, a Student t test was used on the basis of the mean of the volunteer median values. To compare both field strengths, we calculated the Pearson correlation coefficient (r)., Results: DW-EPI MR imaging visualized the parotid glands of all volunteers. With 1.5T, the mean ADC before stimulation was 1.12 x 10(-3) mm(2)/s +/- 0.08 x 10(-3) mm(2)/s. After stimulation with lemon juice, the ADC increased to 1.18 x 10(-3) mm(2)/s +/- 0.09 x 10(-3) mm(2)/s. For 3T, the ADC before stimulation was 1.14 x 10(-3) mm(2)/s +/- 0.04 x 10(-3) mm(2)/s, with an increase to 1.17 x 10(-3) mm(2)/s +/- 0.05 x 10(-3) mm(2)/s after stimulation. For both field strengths, the increase in ADC after stimulation was significant (P < .001). High correlations between both field strengths were found pre- and poststimulation (r = 0.955, and 0.936, respectively)., Conclusion: DW-EPI MR imaging allows monitoring of physiologic changes due to oral stimulation of parotid glands by using DW imaging with high correlation between 1.5T and 3T.

Objective: To study the preservation of parotid glands function and relationship between parotid glands function and dose-volume histogram (DVH) in nasopharyngeal carcinoma (NPC) patients treated by intensity modulated radiation therapy (IMRT)., Methods: From August 2002 to December 2004, the excretion index (EI) and uptake index (UI) of parotids in 48 NPC patients underwent radical IMRT was examined by ECT at the beginning, the end of and the 3 months after radiotherapy. The relationship between parotid function (EI and UI) and DVH were analyzed., Results: The mean doses to the contralateral parotid and ipsilateral parotid were 22.8 +/- 4.5 Gy and 31.9 +/- 4.1 Gy, respectively. The symptom of xerostomia was mild at the end of radiotherapy. ECT showed EI of contralateral parotid were 0.35 +/- 0.25, 0.31 +/- 0.24 and 0.33 +/- 0.22 at the beginning, the end of and 3 months after radiotherapy (RT), respectively. UI were 7.12 +/- 3.56, 5.81 +/- 2.25 and 5.72 +/- 2.81 at the same intervals. This shows no statistical difference. The EI and UI of ipsilateral parotid at the completion of radiotherapy declined significantly (0.21 +/- 0.16 and 4.87 +/- 2.45, respectively) compared with those of pre-treatment (0.36 +/- 0.27 and 8.02 +/- 3.89, respectively) (P < 0.05). DVH showed: at the end of RT, the EI was significant difference between mean dose < 26 Gy and > or = 26 Gy group (P = 0.009) and decreased significantly in the group of V25 (the percentages of parotid volume irradiated with < 25 Gy) > or = 50% compared with the group of V25 < 50% (P < 0.01). The UIs were no significant difference in two groups (P > 0.05)., Conclusion: 26 Gy is a threshold dose for the preservation of parotid glands function. There is also a threshold volume irradiated for the preservation of the parotid glands function. Based on the precondition of assuring significant dose to the target volume (PTV), we should reduce the irradiated volume and dose to parotid glands as possible as we can so as to preserve its function.

The complexity of mammalian physiology requires a diverse array of ion channel proteins. This diversity extends even to a single family of channels. For example, the family of Ca2+-activated K channels contains three structural subfamilies characterized by small, intermediate, and large single channel conductances. Many cells and tissues, including neurons, vascular smooth muscle, endothelial cells, macrophages, and salivary glands express more than a single class of these channels, raising questions about their specific physiological roles. We demonstrate here a novel interaction between two types of Ca2+-activated K channels: maxi-K channels, encoded by the KCa1.1 gene, and IK1 channels (KCa3.1). In both native parotid acinar cells and in a heterologous expression system, activation of IK1 channels inhibits maxi-K activity. This interaction was independent of the mode of activation of the IK1 channels: direct application of Ca2+, muscarinic receptor stimulation, or by direct chemical activation of the IK1 channels. The IK1-induced inhibition of maxi-K activity occurred in small, cell-free membrane patches and was due to a reduction in the maxi-K channel open probability and not to a change in the single channel current level. These data suggest that IK1 channels inhibit maxi-K channel activity via a direct, membrane-delimited interaction between the channel proteins. A quantitative analysis indicates that each maxi-K channel may be surrounded by four IK1 channels and will be inhibited if any one of these IK1 channels opens. This novel, regulated inhibition of maxi-K channels by activation of IK1 adds to the complexity of the properties of these Ca2+-activated K channels and likely contributes to the diversity of their functional roles.

Various ClC-type voltage-gated chloride channel isoforms display a double barrel topology, and their gating mechanisms are thought to be similar. However, we demonstrate in this work that the nearly ubiquitous ClC-2 shows significant differences in gating when compared with ClC-0 and ClC-1. To delineate the gating of ClC-2 in quantitative terms, we have determined the voltage (V(m)) and time dependence of the protopore (P(f)) and common (P(s)) gates that control the opening and closing of the double barrel. mClC-2 was cloned from mouse salivary glands, expressed in HEK 293 cells, and the resulting chloride currents (I(Cl)) were measured using whole cell patch clamp. WT channels had I(Cl) that showed inward rectification and biexponential time course. Time constants of fast and slow components were approximately 10-fold different at negative V(m) and corresponded to P(f) and P(s), respectively. P(f) and P(s) were approximately 1 at -200 mV, while at V(m) > or = 0 mV, P(f) approximately 0 and P(s) approximately 0.6. Hence, P(f) dominated open kinetics at moderately negative V(m), while at very negative V(m) both gates contributed to gating. At V(m) > or = 0 mV, mClC-2 closes by shutting off P(f). Three- and two-state models described the open-to-closed transitions of P(f) and P(s), respectively. To test these models, we mutated conserved residues that had been previously shown to eliminate or alter P(f) or P(s) in other ClC channels. Based on the time and V(m) dependence of the two gates in WT and mutant channels, we constructed a model to explain the gating of mClC-2. In this model the E213 residue contributes to P(f), the dominant regulator of gating, while the C258 residue alters the V(m) dependence of P(f), probably by interacting with residue E213. These data provide a new perspective on ClC-2 gating, suggesting that the protopore gate contributes to both fast and slow gating and that gating relies strongly on the E213 residue.

Exocrine cells of the digestive system are specialized to secrete protein and fluid in response to neuronal and/or hormonal input. Although morphologically similar, parotid and pancreatic acinar cells exhibit important functional divergence in Ca(2+) signaling properties. To address whether there are fundamental differences in exocytotic release of digestive enzyme from exocrine cells of salivary gland versus pancreas, we applied electrophysiological and optical methods to investigate spatial and temporal characteristics of zymogen-containing secretory granule fusion at the single-acinar cell level by direct or agonist-induced Ca(2+) and cAMP elevation. Temporally resolved membrane capacitance measurements revealed that two apparent phases of exocytosis were induced by Ca(2+) elevation: a rapidly activated initial phase that could not be resolved as individual fusion events and a second phase that was activated after a delay, increased in a staircaselike fashion, was augmented by cAMP elevation, and likely reflected both sequential compound and multivesicular fusion of zymogen-containing granules. Optical measurements of exocytosis with time-differential imaging analysis revealed that zymogen granule fusion was induced after a minimum delay of approximately 200 ms, occurred initially at apical and basolateral borders of acinar cells, and under strong stimulation proceeded from apical pole to deeper regions of the cell interior. Zymogen granule fusions appeared to coordinate subsequent fusions and produced persistent structures that generally lasted several minutes. In addition, parotid gland slices were used to assess secretory dynamics in a more physiological context. Parotid acinar cells were shown to exhibit both similar and divergent properties compared with the better-studied pancreatic acinar cell regarding spatial organization and kinetics of exocytotic fusion of zymogen granules.

Aquaporin-5 (AQP5), an apical plasma membrane (APM) water channel in salivary glands, lacrimal glands, and airway epithelium, has an important role in fluid secretion. M(3) muscarinic acetylcholine receptor (mAChR)-induced changes in AQP5 localization in rat parotid glands were investigated with immunofluorescence or immunoelectron microscopy, detergent solubility, and gradient density floatation assays. Confocal microscopy revealed AQP5 localization in intracellular vesicles of interlobular duct cells in rat parotid glands and AQP5 trafficking to the APM 10 min after injection of the mAChR agonist cevimeline. Conversely, 60 min after injection, there was a diffuse pattern of AQP5 staining in the cell cytoplasm. The calcium ionophore A-23187 mimicked the effects of cevimeline. Immunoelectron microscopic studies confirmed that cevimeline induced AQP5 trafficking from intracellular structures to APMs in the interlobular duct cells of rat parotid glands. Lipid raft markers flotillin-2 and GM1 colocalized with AQP5 and moved with AQP5 in response to cevimeline. Under control conditions, the majority of AQP5 localized in the Triton X-100-insoluble fraction and floated to the light-density fraction on discontinuous density gradients. After 10-min incubation of parotid tissue slices with cevimeline or A-23187, AQP5 levels decreased in the Triton X-100-insoluble fraction and increased in the Triton X-100-soluble fraction. Thus AQP5 localizes in the intracellular lipid rafts, and M(3) mAChR activation induces AQP5 trafficking to the APM with lipid rafts via intracellular Ca(2+) signaling and induces AQP5 dissociation from lipid rafts to nonrafts on the APM in the interlobular duct cells of rat parotid glands.

Using patch-clamp and molecular biological techniques, we identified and characterized membrane currents most likely generated by an electrogenic Na+-HCO3- cotransporter (NBCe) in acutely dissociated bovine parotid acinar (BPA) cells. When BPA cells were dialysed with a N-methyl-D-glucamine (NMDG)-glutamate-rich pipette solution, switching a Na-glutamate-rich, nominally HCO3--free bath solution to the one containing 25 mM HCO3-, but not Cl-, elicited a whole-cell current with a linear current-voltage relation. The HCO3- evoked current was abolished by total replacement of extracellular Na+ (Na+o) with NMDG or by 0.5 mM 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid (DIDS), and was only partially supported by Li+o, but not by K+o, Cs+o, and cholineo. The reversal potential shift of DIDS (0.5 mM)-sensitive current induced by a change of [Na+]o corresponded to an apparent coupling ratio of HCO3- to Na+ of 2. RT-PCR analysis showed the presence of transcripts of NBCe1-B, but not NBCe1-A in BPA cells. Electrophysiological and pharmacological properties of whole-cell currents recorded from HEK293 cells transfected with the NBCe1-B, which was cloned from BPA cells resembled those of the native currents. Non-invasive measurements of membrane potential changes in the cell-attached patch configuration indicated that an NBCe activity is present in intact unstimulated BPA cells bathed in a 25 mM HCO3--containing solution. Collectively, these results not only suggest that an NBCe is present, functional and may be mediated, at least in part, by NBCe1-B in BPA cells, but also provide the first electrophysiological characterization of transport properties of NBCe expressed in native exocrine glands.

Upon continuous stimulation, the pore of the monovalent cation-selective P2X7 receptor (P2X7R) expands to accommodate large molecules such as N-methyl-D-glucamine (NMDG+). How the change in P2X7R permeability is regulated is not known. Here we report that extracellular Cl- (Cl-(o)) regulates the outward current, whereas extracellular Na+ (Na+(o)) regulates the inward current of large molecules by P2X7Rs. The P2X7R-mediated current was measured in parotid acinar and duct cells of wild type and P2X7R-/- mice and in HEK293 cells expressing the human or mouse P2X7R isoforms. In symmetrical NaCl, triethylammonium chloride, and NMDG+ chloride solutions, the P2X7R current followed a linear current/voltage relationship. In symmetrical NaCl, removal of Cl-(o) reduced the inward Na+ current by approximately 35% and the outward Na+ current by only 10%. By contrast, in the absence of Na+(i) and the presence of Na+(o) or NMDG+(o), the removal of Cl-(o) reduced the inward Na+ or NMDG+ currents by 35% but the outward NMDG+ current by >95%. The effect of Cl-(o) was half-maximal at approximately 60 mm. Reducing Cl-(i) from 150 to 10 mm did not reproduce the effects of Cl-(o). All currents were eliminated in P2X7R-/- cells and reproduced by expressing the P2X7Rs in HEK cells. These findings suggest that Cl-(o) primarily regulates the outward P2X7R current of large molecules. When cells dialyzed with NMDG+ were stimulated in the presence of Na+(o), subsequent removal of Na+(o) resulted in a strongly outward rectifying NMDG+ current, indicating maintained high selectivity for Na+ over NMDG+. During continuous incubation in Na+-free medium, the permeability of the P2X7Rs to NMDG+ gradually increased. On the other hand, when the cells were incubated in symmetrical NMDG+ and only then stimulated with ATP, the NMDG+ current by P2X7Rs followed a linear current/voltage relationship and did not change with time. These findings suggest that the P2X7R has a "Na+(o) memory" and that Na+(o) regulates the inward permeability of P2X7Rs to large molecules. The novel regulation of P2X7R outward and inward permeability to large molecules by Cl-(o) and Na+(o), respectively, may have an important protective function, particularly in secretory epithelial cells.