Preparation of Soluble Mucin Solutions from the Salivary Glands.

Studying salivary gland mucins is important for elucidating the pathogenesis of salivary gland diseases, including tumors and xerostomia, and developing diagnostic methods for them. Classic methods for isolating mucins from salivary glands require sacrificing several animals to obtain sufficient quantities of mucin and are time-consuming. Supported molecular matrix electrophoresis (SMME) was used to characterize mucins and their glycans. With this method, mucins can be analyzed within 2 days using less than 100 mg of tissue and without using expensive equipment, such as an ultracentrifuge. This chapter describes a method for preparing mucin solutions for SMME analysis of salivary gland mucins.

Expression and localisation of MUC1 modified with sialylated core-2 O-glycans in mucoepidermoid carcinoma.

Mucoepidermoid carcinoma (MEC) is the most frequent of the rare salivary gland malignancies. We previously reported high expression of Mucin 1 (MUC1) modified with sialylated core-2 O-glycans in MEC by using tissue homogenates. In this study, we characterised glycan structures of MEC and identified the localisation of cells expressing these distinctive glycans on MUC1. Mucins were extracted from the frozen tissues of three patients with MEC, and normal salivary glands (NSGs) extracted from seven patients, separated by supported molecular matrix electrophoresis (SMME) and the membranes stained with various lectins. In addition, formalin-fixed, paraffin-embedded sections from three patients with MEC were subjected to immunohistochemistry (IHC) with various monoclonal antibodies and analysed for C2GnT-1 expression by in situ hybridisation (ISH). Lectin blotting of the SMME membranes revealed that glycans on MUC1 from MEC samples contained α2,3-linked sialic acid. In IHC, MUC1 was diffusely detected at MEC-affected regions but was specifically detected at apical membranes in NSGs. ISH showed that C2GnT-1 was expressed at the MUC1-positive in MEC-affected regions but not in the NSG. MEC cells produced MUC1 modified with α2,3-linked sialic acid-containing core-2 O-glycans. MUC1 containing these glycans deserves further study as a new potential diagnostic marker of MEC.

Characterization of tumor-associated MUC1 and its glycans expressed in mucoepidermoid carcinoma.

Mucoepidermoid carcinoma (MEC) is one of the most frequently misdiagnosed tumors. Glycans are modulated by malignant transformation. Mucin 1 (MUC1) is a mucin whose expression is upregulated in various tumors, including MEC, and it has previously been investigated as a diagnostic and prognostic tumor marker. The present study aimed to reveal the differences in the mucin glycans between MEC and normal salivary glands (NSGs) to discover novel diagnostic markers. Soluble fractions of salivary gland homogenate prepared from three MEC salivary glands and 7 NSGs were evaluated. Mucins in MEC and NSGs were separated using supported molecular matrix electrophoresis, and stained with Alcian blue and monoclonal antibodies. The glycans of the separated mucins were analyzed by mass spectrometry. MUC1 was found in MEC but not in NSGs, and almost all glycans of MUC1 in MEC were sialylated, whereas the glycans of mucins in NSGs were less sialylated. The core 2 type glycans, (Hex)2(HexNAc)2(NeuAc)1 and (Hex)2(HexNAc)2(NeuAc)2, were found to be significantly abundant glycans of MUC1 in MEC. MEC markedly produced MUC1 modified with sialylated core 2 glycans. These data were obtained from the soluble fractions of salivary gland homogenates. These findings provide a basis for the utilization of MUC1 as a serum diagnostic marker for the preoperative diagnosis of MEC.

Temporary Closure of Inpatient Ward due to Mumps Virus Reinfection in Elderly Patient.

We report a case of suspected reinfection with the mumps virus in an elderly patient which resulted in temporary closure of an inpatient ward. A 65-year-old man with colorectal cancer was admitted to the digestive surgery ward at our hospital to undergo a stoma closure operation. He was subsequently referred to our department with right swelling in the preauricular region on postoperative day 4. The swelling subsided within a few days, and the patient was discharged. A serum titer test revealed a high level of antibodies to the mumps virus, however. Therefore, staff who had come into close contact with the patient were examined and the decision taken to stop admitting new patients to the ward. When symptoms are detected in a patient has already had mumps, it is important to consider the possibility of reinfection. Furthermore, it is necessary for medical workers to undergo a serum antibody test to the mumps virus and receive a further vaccination if antibody levels are too low to confer immunity.