Immune complexome analysis reveals the presence of immune complexes and identifies disease-specific immune complex antigens in saliva samples from patients with Sjögren's syndrome.

Sjögren's syndrome (SS) is a chronic autoimmune disease that mainly damages the salivary and lacrimal glands. Immune complex (IC) formation triggers local inflammation through IC deposition and decreased antigen function. Some ICs can leak from the lesion and into the saliva, but no salivary ICs have been reported to date. We used immune complexome analysis to comprehensively identify antigens incorporated into IC (IC-antigens) in saliva samples from patients with SS (n = 9) or with xerostomia (n = 7). Neutrophil defensin 1 (67%), small proline-rich protein 2D (67%), myeloperoxidase (44%), neutrophil elastase (44%), cathepsin G (33%), nuclear mitotic apparatus 1 (33%) and phosphatidylinositol 4-phosphate 3-kinase C2 domain-containing subunit gamma (33%) were identified as new IC-antigens specifically and frequently detected in the saliva of SS patients. Of these, neutrophil defensin 1, myeloperoxidase, neutrophil elastase and cathepsin G are neutrophil intracellular proteins, which suggests that repeated destruction of neutrophils due to abnormal autoimmunity may be involved in the pathogenesis of SS. We also analyzed serum samples from three SS patients. There was little overlap of IC-antigens between two of the samples (fewer than 30% of the IC-antigens in the saliva samples), suggesting that many ICs are formed locally and independently of the circulation. In addition, we found that four SS-specific salivary antigens show sequence homology with several proteins of oral microbiomes but no antigen has homology with Epstein-Barr virus proteins. The homology between some IC-antigens and oral microbiome proteins may indicate the impact of oral infection on local autoimmunity through molecular mimicry theory.

Simultaneous determination of phenolic xenoestrogens by solid-phase extraction and high-performance liquid chromatography with fluorescence detection.

A highly sensitive and selective method for simultaneous determination of some hydroxyl group-containing endocrine disruptors, including bisphenol A (BPA), bisphenol B (BPB), bisphenol E (BPE), bisphenol F (BPF) and 4-nonylphenol (4-NP), was developed. The method consists of precolumn derivatization of the analytes, solid-phase extraction (SPE) and subsequent chromatographic analysis by high-performance liquid chromatography (HPLC) with fluorescence detection. 4,4'-Cyclohexylidenebisphenol (BPZ) was used as an internal standard. Derivatization was carried out using 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl) as a label. Parameters of the derivatization reaction (temperature, time, concentration of reagent, stability, etc.) and of the solid-phase extraction (recovery, solvent, etc.) were studied in detail. Detection limits of compounds studied in standard solutions ranged from 0.08-1.3 ppb (ng/ml). The proposed method was successfully applied to plastic samples; BPA was found in both polycarbonate and polyvinyl chloride plastics, while 4-NP was found in plastics made of polyvinyl chloride and another polymer.

A case of homicidal choking mistaken for suicide.

This report presents an autopsy case of the homicidal choking of an adult, who died as a result of tissue paper being thrust into his mouth. An in-patient (a 29-year-old Japanese man) at a mental hospital was choked to death by another male in-patient, his roommate, who thrust a large amount of tissue paper into his mouth after rendering him slightly unconscious by cervical compression. At the time of discovery, this mental patient was believed to have committed suicide by inserting tissue paper into his own mouth. Autopsy findings and re-investigation of the case revealed that the subject had been murdered. This is a very rare case of choking used as a method of homicide.

Comparison of plastic prostheses and self-expandable metallic stents in the treatment of malignant esophageal stenosis.

OBJECTIVE: We compared the efficacy of plastic prostheses and self-expandable metallic stents in the treatment of malignant esophageal stenosis and/or fistula. SUBJECTS AND METHODS: Subjects were 31 patients with esophageal cancer, 4 with esophagotracheal fistula, and 1 with esophageal stenosis. A plastic prosthesis was inserted in 15 patients (group A) and a self-expandable metallic stent in 21 patients (group B). We evaluated food ingestion improvement, effectiveness, complications, mean survival, food ingestion duration, the percentage of food ingestion capability in total survival time, and inhospital mortality. RESULTS: No differences were seen in food intake improvement or in the effectiveness between groups, whereas fatal complications were higher in Group A. No significant differences were seen in mean survival, food ingestion duration, percentage of food ingestion capability, or inhospital mortality between groups. CONCLUSION: We concluded that a self-expandable metallic stent was safer than a plastic prosthesis because of fewer serious complications such as bleeding, and recommended the use of metallic stents in the treatment of malignant esophageal stenosis and/or fistula.

[Histopathological and immunohistochemical studies on mucous cysts].

The present study investigated the histopathology, histochemistry of mucopolysaccharides, and immunohistochemistry of oral mucous cysts. The materials were obtained from ninety cases that were histopathologically diagnosed as oral mucous cysts at the Department of Oral Pathology, Meikai University School of Dentistry. Mucopolysaccharide staining was done with PAS, alcian blue (AB, pH 2.5) and high iron diamine (HID). Immunohistochemical studies were focused on secretory component (SC), lactoferrin (Lf), alpha-amylase (Am), IgA, lysozyme (Ly), and keratin (Kr). The following results were obtained: 1. Histopathological findings. (1) Retention and/or retention-like type cysts occurred in was twenty-six cases and the extravasation type in sixty-four cases. (2) Cases showing epithelial lining of the cystic wall were only eight in number, and many cystic walls were contained granulation tissue (fifty cases). (3) As for inflammation of the cystic wall, the degree was slight, and infiltrated cells were mainly macrophages (so-called mucinophages) and lymphocytes. (4) Regarding adjoining salivary glands, acinar cells showed atrophic changes, and hypertrophy of mucous acinar cells was evident. Many ducts showed dilatation, and stromal connective tissue showed fibrosis and hyalinization. 2. Histochemical findings on mucopolysaccharides. (1) Mucous materials in cystic cavity, mucous acinar cells, and secretory materials in ductal lumens were intensely stained by PAS and AB. But stainability with AB was less than that with PAS staining. Serous acinar cells and ductal epithelium were negative to PAS and AB staining. (2) Stainability of the above with HID was less than at with PAS or AB. Cystic walls were not stained by HID. Mucous acinar cells reactive with HID were intensely stained, but the number of the positive cells was limited when compared with the numbers of PAS-and AB-positive cells. 3. Immunohistochemical findings. (1) As for mucous materials in the cystic cavity, reactions for Sc, Am, IgA, and Ly were all positive, whereas those for Lf and Kr were negative. (2) Staining of cystic walls was generally weak: the walls were negative for IgA, Ly and Kr showed and borderline or slightly positive for Sc, Lf, and Am. (3) Mucous acinar cells were negative for all markers examined in this study, but serous acinar cells and/or demilunes were markedly positive for Sc, Ly, and Am. (4) In ductal epithelial cells, Ly and Kr were negative, but IgA was borderline or slightly positive. Sc and Am gave intensely positive staining. (5) Secretory materials in ductal lumens, Sc were intensely positive for Lf, and Ly; slightly or moderately positive for IgA; and slightly positive or negative for Am.(ABSTRACT TRUNCATED AT 400 WORDS)

[Relationship between occlusal interference and hearing ability].

Clicking sounds are one of the symptoms that appear in the temporomandibular joint dysfunction at a high rate. But they are sometimes found in healthy subjects also, so that clicking sounds are seldom recognized as an pathologic condition. Study reports on clicking sounds in the past concentrated mainly on their cause, process of their appearance and their spectral properties. These studies were, however, all based on the understanding that clicking sounds were a kind of symptom or signals that accompanied the unbalanced jaw relationship. They did not examine pathologic conditions or functional change caused by the very existence of clicking sounds. This author has been reporting on the influence of clicking sounds on hearing abilities. This time the author considered it necessary to study the cause for rise of the hearing level of the clicking subjects in relation to the unbalanced positions of their jaws. The author therefore examined the relationship between the phases of occlusal contact and hearing abilities of the clicking subjects. From the results, we obtained the following conclusions: 1. As for the frequency of appearance of clicking sounds, 28 out of the 175 healthy subjects (aged either 21 or 22) had clicking sounds on one side, accounting for 16.0%, at the hazard rate of 1%. There was no differences between the sexes and between the right and left sides. 2. Of the 37 subjects, the cases in which the centric position fall on the intercuspal position were only three among the 17 non-clicking subjects (Group I), accounting for 17.6% and none, among the 20 clicking subjects.(ABSTRACT TRUNCATED AT 250 WORDS)

Morphological modifications during long-term survival of Meckel's cartilage hypertrophic chondrocytes transplanted in the mouse spleen.

To examine the ultimate fate of hypertrophic chondrocytes, Meckel's cartilage bars from 18-day-old mouse embryos were transplanted into isogenic mouse spleen for 3, 7, 14 and 21 days and observed at the light and electron microscopic levels. The midportions of these Meckel's cartilage bars were used as explants; they were characterized by many hypertrophic chondrocytes containing euchromatic round nuclei, a large amount of glycogen particles, and some vacuoles. Grafted cartilage adapted well to the splenic tissue, showing intense metachromasia around the territorial matrix. Ultrastructural observations indicated that the number of large vacuoles and glycogen aggregates in the hypertrophic cells became markedly reduced with grafting time, whereas the Golgi apparatus and rough endoplasmic reticulum were well-developed. Needle-like crystals showing initial apatite deposition appeared in association with matrix vesicles; these proliferated as time elapsed after transplantation. On day 14 after transplantation, cells displaying such various structural features as pyknotic nuclei, large vacuoles, and cytoplasmic shrinkage were noted in addition to intact hypertrophic chondrocytes. Following resorption of the calcified cartilage by multinucleated giant cells, many osteoblasts appeared along the border of the calcified matrix. Some remaining hypertrophic cells in the calcified matrix had transformed into osteocyte-like cells. On day 21, the resorbed area of the calcified cartilage was invaded by many blood vessels. Hypertrophic chondrocytes, now exposed from cellular lacunae, and the osteocyte-like cells in the calcified matrix displayed involutional changes. The present study showed that, although the hypertrophic chondrocytes in Meckel's cartilage essentially underwent regressive changes, they retained the ability to stimulate endochondral ossification within the microenvironment of the spleen. In addition, some of these cells were transformed into osteocyte-like cells.

High-performance liquid chromatography with peroxyoxalate chemiluminescence detection of bisphenol A migrated from polycarbonate baby bottles using 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride as a label.

A highly sensitive and selective high-performance liquid chromatographic method with peroxyoxalate chemiluminescence detection for the determination of bisphenol A at sub-ppb levels is described. Bisphenol A was derivatized with 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride and the excess unreacted reagent was removed by a simple solid-phase extraction procedure with recoveries of approximately 60%. The separation was carried out isocratically on an ODS column and the derivatized bisphenol A was detected by peroxyoxalate chemiluminescence. A mixture of bis[2-(3,6,9-trioxadecanyloxycarbonyl)-4-nitrophenyl]oxalate (0.6 mM) and hydrogen peroxide (25.0 mM) dissolved in acetonitrile was used as a chemiluminescence reagent solution with a mixture of imidazole-HNO3 buffer (40.0 mM, pH 7.0): acetonitrile (17:83, v/v) as a mobile phase. The linear standard curve was obtained over the range from 0.57 (2.5) to 22.8 (100) ppb (nM) (r=0.996) with a detection limit of 0.38 ppb (2.8 fmol on column) at a signal-to-noise ratio of 3. The method was successfully applied to the determination of bisphenol A in hot water in contact with commercially available baby bottle samples.