Expression of interferon-gamma-inducible protein-10 in human endometrial stromal cells

Human endometrial stromal cells (ESC) can produce a variety of chemokines, especially after inflammatory stimulation. Interferon-gamma-inducible protein-10 (IP-10) is a potent chemoattractant for lymphocytes, and belongs to the family of non-ELR CXC chemokines. The expression of IP-10 in ESC after stimulation with interferon-gamma (IFN-gamma), interleukin-1 beta (IL-1 beta), tumour necrosis factor-alpha (TNF-alpha), or lipopolysaccharide (LPS) was evaluated using an enzyme-linked immunosorbent assay and Northern blot analysis. A small amount of IP-10 protein was detected in the culture media of unstimulated ESC. The expression of IP-10 mRNA was detected in ESC. IFN-gamma, IL-1 beta, TNF-alpha and LPS significantly stimulated the expression of IP-10 mRNA and protein in ESC. These results suggest that the production of IP-10 by ESC is regulated by inflammatory mediators. The modulation of IP-10 concentrations in the local environment may contribute to the normal and pathological processes of human reproduction by regulating leukocyte trafficking in the endometrium.     

Immunoregulatory defects of V alpha 24V+ beta 11+ NKT cells in development of Wegener's granulomatosis and relapsing polychondritis

The frequency of either CD4(-)8(-) (double negative; DN) or CD4(+) V alpha 24(+)V beta 11(+) NKT cells, the expression of CD1d and the binding of CD1d-tetramer loaded with alpha-galactosylceramide (alpha-GalCer) to NKT cells were analysed in peripheral blood mononuclear cells (PBMCs) of patients with Wegener's granulomatosis (WG), relapsing polychondritis (RP) and healthy subjects (HS). DN and CD4(+) V alpha 24(+)V beta 11(+) NKT cells as well as CD1d-alpha-GalCer tetramer-positive NKT cells, were significantly decreased in number in both WG and RP patients compared to those from HS. When cytokine profiles were analysed in these PBMCs upon stimulation with phorbol ester and calcium ionophore, CD4(+) T cells from patients with WG and RP exhibited a Th1 bias, whereas CD4(+) NKT cells from WG patients in remission showed a Th2 bias. These findings suggest that NKT cells (especially CD4(+) NKT cells) play a regulatory role in Th1 autoimmunity in patients with WG and RP. The reduction in NKT cell counts appears to be associated with the low responsiveness to alpha-GalCer. The dysfunction of NKT cells to recognize ligands such as alpha-GalCer may also contribute to the defects observed in NKT cells from WG and RP patients.     

Immunohistochemical analysis of centromere protein F expression in buccal and gingival squamous cell carcinoma

Centromere protein F (CENP-F) expression (localization and characteristics) in relation to tumor clinicopathological parameters was immunohistochemically examined and evaluated in 47 archival biopsy specimens of buccal and gingival squamous cell carcinomas (SCC). Centromere protein F expression was detected in 79% of the samples. An increase in the labeling index (LI) with WHO grading was obtained ( P  < 0.05). Correlations were obtained between the CENP-F LI and tumor size ( P  < 0.05). Immunoelectron microscopy showed CENP-F nuclear staining as punctate or fine dots. The present study shows that CENP-F expression and detection of a more specific cell subpopulation presents a theoretical advantage for the analysis of the precise cell cycle of G₂ to M cells, compared to Ki-67.     

Direct in vivo protein transduction into a specific restricted brain area in rats

Attempts at protein transduction into specific restricted brain areas have remained unsuccessful. We attempted targeted, direct in vivo protein transduction by microinjecting beta-galactosidase (beta-gal) with hemagglutinating virus of Japan envelope (HVJ-E) vector into the rat nucleus tractus solitarius (NTS). The medulla oblongata including the NTS was removed 6h post-injection and cryostat sections were histochemically stained to detect beta-gal enzymatic activity. beta-gal-positive cells were present in these sections as was beta-gal activity determined by colorimetric analysis. beta-gal-positive cells were not present in the rats microinjected only beta-gal protein without HVJ-E vector. Our findings suggest that direct in vivo protein transduction into specific restricted brain areas is possible. The type of targeted delivery system we present may have wide applications in the administration of therapeutic proteins to the central nervous system.     

The effect of vitamin A depletion on antigen-stimulated trapping of peripheral lymphocytes in local lymph nodes of rats

The effect of vitamin A depletion on antigen-stimulated trapping of peripheral lymphocytes in lymphatic organs was studied in rats. Distribution of [3H]-uridine-labelled syngenic peripheral lymphocytes was quantified by assaying radioactive content of brachial and axillary lymph nodes, spleen and liver of normal and vitamin A-depleted F344/Ducrj rats immunized with sheep red blood cells. Localization of labelled cells in the ipsilateral brachial lymph nodes of the normal rats was stimulated by three times upon immunization with sheep erythrocytes as compared with the contralateral nodes. Recruitment of cells in axillary lymph nodes, spleen and liver was not significantly different from non-immunized values. The vitamin A-depleted rats exhibited marked deterioration in antigen-stimulated trapping of labelled cells in the draining brachial lymph nodes. These results suggest that this effect of vitamin A depletion is due to derangement of integrity of lymphocyte-trapping mechanism in the draining lymph nodes and not to any change in nature of lymphocytes per se.     

Cellular Adherence, Glucosyltransferase Adsorption, and Glucan Synthesis of Streptococcus mutans AHT Mutants

Streptococcus mutans AHT mutants M1, M2, and M13 failed to adhere to a glass surface, whereas mutants M9 and M35 exhibited decreased and increased adherence, respectively, as compared with the parent strain, when grown in sucrose broth. Extracellular glucosyltransferase prepared from glucose-grown cultures of the adherent strains (wild type, M9, and M35) induced adherence of heat-killed cells of the homologous and heterologous streptococcal strains as well as of Escherichia coli K-12 and uncoated resin particles. The glucosyltransferase was adsorbed on all the streptococcal cells and glucan-coated resins, but not on E. coli cells and the uncoated resins. Glucosyltransferase from the nonadhering mutants (M1, M2, M13) neither was significantly adsorbed on nor induced adherence of any of the cells and resins. Cell-free enzymes from the glucose-grown adherent strains produced water-soluble and water-insoluble glucans, whereas those from the nonadhering mutants produced only water-soluble glucans. Small amounts of alkali-soluble, cell-associated glucan were recovered from the sucrose-grown nonadhering mutants. Thus, the relative proportions of glucosyltransferase isozymes elaborated by the S. mutans mutants, insofar as they affect the physico-chemical properties of the glucans produced, seem to determine the adherence abilities of the cells. The adsorption of glucosyltransferase on glucan molecules on the cell surface is not required for the adherence of S. mutans, but de novo glucan synthesis is important in the adherence process.     

Modelling HA protein-mediated interaction between an influenza virus and a healthy cell: pre-fusion membrane deformation.

We present a mathematical model for pre-fusion interaction between an influenza virus and a healthy cell. Our model describes the role played by hemagglutinin (HA) protein clusters in bringing the viral membrane into close contact with the host cell membrane as a first step of the fusion process between the two membranes. The viral membrane is modelled as a lipid bilayer with bending rigidity. Using the calculus of variations, we compute the deformation of the viral membrane under the influence of HA protein clusters. Our numerical results support the hypothesis of dimple formation in the fusion site proposed in the literature. The asymmetric nature of the protein molecules due to various reasons such as tilting is the primary cause for the dimple formation. We discuss the effects of spontaneous curvature, the protein cluster radius, fusion-site size and the bending moment exerted by the protein cluster. We also examine the effects of membrane tension and the presence of a host cell on the dimple shape. Our results support previous experimental observations.     

Bone bonding behavior of three kinds of apatite containing glass ceramics

We have produced three kinds of apatite-containing glass ceramics of the same chemical composition: MgO (4.6), CaO (44.9), SiO2 (34.2), P2O5 (16.3), CaF2 (0.5) (in weight ratio). They contain different crystal combinations and have different mechanical properties. The first glass ceramic (A-GC) was prepared by heating a glass plate to 870 degrees C. It contains only oxy- and fluoroapatite (35 wt%). The second glass ceramic (A-W-GC), and the third (A-W-CP-GC), were prepared by heating glass powder compacts to 1050 degrees C and 1200 degrees C, respectively. A-W-GC contains oxyapatite and fluoroapatite (Ca10(PO4)6(O,F2] (35 wt%) and beta-wollastonite (40 wt%). A-W-CP-GC contains oxyapatite and fluoroapatite (20 wt%), beta-wollastonite (CaO X SiO2) (55 wt%), and beta-whitlockite (3CaO X P2O5) (15 wt%). The bending strengths of A-GC, A-W-GC, and A-W-CP-GC were 88MPa, 178MPa, and 213MPa, respectively, in air. Rectangular ceramic plates (15mm X 10mm X 2mm) were implanted into a rabbit tibia. Ten and 25 weeks after implantation, the segment of tibia with implant was excised for examination. The segment was held by a special jig and the traction breaking load (failure load) was measured by an autograph. A-GC showed a lower load than A-W-GC and A-W-CP-GC. The loads for A-W-GC and A-W-CP-GC were almost equal. The failure loads did not change significantly between 10 and 25 weeks for any of the materials. The interface was examined by Giemsa surface staining, contact micro-radiography, and SEM-EPMA. Giemsa surface staining and CMR revealed direct bonding between the materials and the bone for all the three materials. SEM-EPMA showed that Si and Mg content decreased, Ca content did not change, and P content increased at the reaction zone between all three glass ceramics and bone. This was observed at 10 weeks, as well as at 25 weeks, after implantation. The reaction zone was narrowest with A-GC, wider with A-W-GC, and widest with A-W-CP-GC.     

Inhibitory effect of oral administration of Lactobacillus casei on 3-methylcholanthrene-induced carcinogenesis in mice

The present study was designed to determine whether tumor induction by 3-methylcholanthrene (MC), a carcinogenic hydrocarbon, can be inhibited by oral administration of Lactobacillus casei strain Shirota (LC). C3H/HeN mice were divided into four groups and assigned to the following treatments: treated with MC and given control or LC-containing diet; treated with vehicle only and given control or LC-containing diet. MC (1 mg) was injected intradermally at 7 weeks of age and the tumor incidence was monitored; LC was mixed into a diet at a concentration of 0.05% (w/w) and the diet was fed from the day of MC injection throughout the study. Spleen cells were analyzed for the immune parameters at 12 and 16 weeks after the MC injection. Oral feeding of mice with LC reduced tumor incidence (P < 0.05). MC treatment lowered the in vitro response to concanavalin A (Con A) of spleen cells, the secretion of interleukin-2 in spleen cell culture after stimulation of the cells with Con A and the proportions of CD3⁺, CD4⁺ and CD8⁺ splenic cells. However, the analysis of the spleen cells obtained from the mice treated with MC and given the LC-containing diet revealed that these disrupted host immune parameters were maintained at the level of normal controls. These results suggest that oral feeding of mice with LC inhibits MC-induced tumorigenesis by modulating the disrupted host immune responses during MC carcinogenesis. Received: 14 April 1999