Metabolomics analysis of saliva from patients with primary Sjögren's syndrome.

The recent development of salivary proteomics has led to the identification of potential biomarkers for diagnosing patients with primary Sjögren's syndrome (pSS). Here we sought to identify differentially produced salivary metabolites from pSS patients and healthy controls (HCs) that might be used to characterize this disease. We obtained salivary samples from 12 female pSS patients (mean age 44.2 ± 13.01) and 21 age-matched female HCs. The metabolite profiles of saliva were analysed by gas chromatography-mass spectrometry. The total metabolite levels in each of the samples were calculated and compared across the study participants. A total of 88 metabolites were detected across the study samples, 41 of which were observed at reduced levels in the samples from pSS patients. Principal component analysis (PCA) revealed a loss in salivary metabolite diversity in the pSS patient samples compared to the HC samples. The reduced presence of glycine, tyrosine, uric acid and fucose, which may reflect salivary gland destruction due to chronic sialoadenitis, contributed to the loss of diversity. Comparative PCA of the pSS patients revealed the presence of two subpopulations based on their metabolite profiles, and these two subpopulations showed a significant difference in the prevalence of major salivary glanditis (P = 0.014). In this study, we found that the salivary metabolite profile of pSS patients was less diverse than that of HCs and that the metabolite profiles in pSS patients were affected by the presence of major salivary glanditis.

Exogenous thrombospondin stimulates the proliferation of non-thrombospondin-producing cells.

The effects of thrombospondin (TSP) on the proliferation of two different human carcinoma cell lines (KIM-1 and CW-2) were investigated. The characterization of these two carcinoma cells by immunohistochemistry using anti-TSP antibody and anti-TSP-receptor antibody showed that the KIM-1 had TSP-receptors and TSP, while the CW-2 had only TSP-receptors. The addition of exogenous TSP (10 or 20 microg/ml) to culture medium stimulated the cell proliferation of CW-2 but not that of KIM-1. The cell count for CW-2 was increased dosage-dependently from 10.3 0.6x104/ml at zero TSP concentration to 12.9 0.6x104/ml at 10 microg/ml TSP concentration and to 14.7 0. 4x104/ml at 20 microg/ml TSP (each p<0.0001). In conclusion, though TSP promoted the proliferation of non-TSP-producing cells, it did not promote proliferation of TSP-producing cells. Therefore, it is predicted that TSP was already at saturated activity concentration in the TSP-producing cell line (KIM-1).

C-CAM expression in the developing rat central nervous system.

C-CAM, a transmembrane glycoprotein belonging to the immunoglobulin superfamily, can mediate intercellular adhesion by homophilic, Ca(2+)-independent binding. Immunohistochemical analysis of adult rat tissues has demonstrated that C-CAM is expressed in various epithelia, vessel endothelia, and hematopoietic cells. By molecular cloning and sequence analysis several isoforms differing both in the extracellular and the cytoplasmic domains have been found. Here we have analyzed the expression of C-CAM in the developing rat central nervous system. No neuronal expression was observed, but biochemical and immunohistochemical analyses demonstrated that C-CAM becomes expressed in the microvessels from embryonic day E-13; the intensity of the staining increased through day E-15 and then gradually decreased during the perinatal and early postnatal period. The expression of C-CAM in the walls of the microvessels was confirmed by in situ hybridization. Immunoelectron microscopy showed that C-CAM was localized both to the abluminal surface of the endothelial cells and to cellular processes of primordial pericytes where these two cell types are in contact with each other. No staining was found on the luminal endothelial cell surfaces or inter-endothelial cell contact areas. During the perinatal period C-CAM also became expressed on the opposite side of the pericytes and on other cells, possibly astrocytes, in contact with these areas of the pericytes. These observations suggest that C-CAM may be involved in heterotypic, homophilic adhesion between endothelial cells, pericytes and astrocytes, and in maturation of the vessel walls.

Detection of passage and absorption of chicken egg yolk immunoglobulins in the gastrointestinal tract of pigs by use of enzyme-linked immunosorbent assay and fluorescent antibody testing.

Chicken egg yolk IgG can be absorbed and transferred as efficiently as colostral antibodies in the blood of neonatal pigs. Egg yolk IgG has a half-life of 1.85 days in newborn pig serum. This is shorter than the reported half-life (12 to 14 days) of homologous IgG in serum of pigs. Similar to colostral antibodies, egg yolk IgG absorption from intestine ceased at about 34 hours of age, after a logarithmic decrease in absorption rate from birth. Egg yolk IgG absorption inhibition time in the gastrointestinal tract took 1.73 hours to decrease by half. Egg yolk IgG was protective against experimentally induced diarrhea in pigs when it was administered at high dose, and multiple dosing was instituted. Adverse effects were not observed when chicken egg yolk IgG was administered orally to pigs.

[Analysis of nuclear DNA heterogeneity of the hepatocellular carcinoma (HCC)].

To evaluate the objective proliferative activity in HCC nuclear DNA contents were measured by means of microspectrophotometry and at the same time the immunohistochemical technique using anti-PCNA antibody was employed. Surgically resected 26 HCCs were analyzed in terms of cell proliferative activity and regional heterogeneity. The analysis was performed by immunohistochemical demonstration of PCNA and pathologic histochemical study in formalin-fixed, paraffin-embedded specimens and cytophotometric measurements of nuclear DNA contents in fresh specimens. The results were as follows. 1) Nine HCCs showed regional ploidy heterogeneity. 2) PCNA labeling index and histological grade of the marginal area was much higher than that of the central area. From these results, we concluded that in the process of the HCC progression proliferative activity was decreased in the central area and was not decreased in the marginal area.

Immunocytochemical discovery of the 22- to 23-Kd subunit of cytochrome b558 at the surface of human peripheral phagocytes.

A monoclonal antibody raised against cytochrome b558 reacted specifically with the 22- to 23-Kd protein, the small subunit of this cytochrome. Cytochemical studies showed that the epitope was located on the surfaces of human neutrophils and monocytes. The small subunit of cytochrome b558, therefore, was expressed at least in part on the outer surface of these cells.

Passive protective effect of chicken egg yolk immunoglobulins against experimental enterotoxigenic Escherichia coli infection in neonatal piglets.

Passive protection of neonatal piglets against fatal enteric colibacillosis was achieved with powder preparations of specific antibodies against K88, K99, and 987P fimbrial adhesins of enterotoxigenic Escherichia coli. The antibody powders were obtained by spray drying the water-soluble protein fraction of egg yolks from immunized hens after the lipid components were precipitated with an aqueous dispersion of acrylic resins (Eudragit L30D-55; Rohm pharma). The anti-K88, -K99, and -987P antibody preparations reacted specifically against the corresponding fimbrial antigens in an enzyme-linked immunosorbent assay. The orally administered antibodies protected in a dose-dependent fashion against infection with each of the three homologous strains of E. coli in passive immunization trials with a colostrum-deprived piglet model of enterotoxigenic E. coli diarrhea. Scanning electron microscopy revealed adherence of enterotoxigenic E. coli in intestinal epithelial surfaces of control piglets, whereas in treated piglets treated with high-titer antibodies, a resistance to bacterial adhesion was observed. An enzyme immunoassay with avidin-biotin complex demonstrated specific local antibody activity in target areas of the small intestines. In vitro, E. coli K88+, K99+, and 987P+ strains adhered equally to porcine duodenal and ileal epithelial cells but failed to do so in the presence of homologous anti-fimbrial antibodies. Absorption of egg yolk antibodies with fimbrial immunosorbent removed the anti-fimbrial antibody fraction and reduced significantly the protective nature of the antibody preparation in a passive immunization experiment, suggesting that anti-fimbrial antibodies were the active components.