Observation and determination of periodontal tissue profile using optical coherence tomography.

BACKGROUND AND OBJECTIVE: Diagnosis is a crucial step in periodontal treatment. The aim of this study was to evaluate the effectiveness of optical coherence tomography (OCT) for observation and determination of periodontal tissue profiles in vivo. MATERIAL AND METHODS: In experiment 1, refractive indices of purified water, porcine gingiva and human gingiva at 1330 nm were determined for the analysis of OCT images of periodontal tissues. In experiment 2, OCT examination was performed in the midlabial apico-coronal plane of mandibular anteriors in 30 Asian volunteers with healthy gingiva. Sulcus depth was measured on intra-oral photographs taken during probing. In the OCT images, the gingival, epithelial and connective tissue thickness, and the position of alveolar bone crest were determined and finally, the biologic width was measured. RESULTS: Refractive indices of purified water, porcine gingiva and human gingiva were 1.335, 1.393 and 1.397, respectively. Cross-sectional images of gingival epithelium, connective tissue and alveolar bone were depicted in real-time. The sulcular and junctional epithelium could be visualized occasionally. Laser penetration and reflection were limited to a certain depth with an approximate maximal imaging depth capability of 1.5 mm and OCT images of the periodontal structure were not clear in some cases. The average maximal thickness of gingiva and epithelium and biologic width at the mandibular anteriors were 1.06 ± 0.21, 0.49 ± 0.15 and 2.09 ± 0.60 mm, respectively. CONCLUSION: OCT has promise for non-invasive observation of the periodontal tissue profile in detail and measurement of internal periodontal structures including biologic width in the anterior region.

Helicobacter pylori CagA protein can be tyrosine phosphorylated in gastric epithelial cells.

Attachment of Helicobacter pylori to gastric epithelial cells induces various cellular responses, including the tyrosine phosphorylation of an unknown 145-kD protein and interleukin 8 production. Here we show that this 145-kD protein is the cagA product of H. pylori, an immunodominant, cytotoxin-associated antigen. Epithelial cells infected with various H. pylori clinical isolates resulted in generation of tyrosine-phosphorylated proteins ranging from 130 to 145 kD in size that were also induced in vitro by mixing host cell lysate with bacterial lysate. When epithelial cells were infected with [(35)S]methionine-labeled H. pylori, a radioactive 145-kD protein was detected in the immunoprecipitates with antiphosphotyrosine antibody or anti-CagA (cytotoxin-associated gene A) antibody. Consistently, the 145-kD protein recognized by the anti-CagA and antiphosphotyrosine antibodies was induced in epithelial cells after infection of wild-type H. pylori but not the cagA::Km mutant. Furthermore, the amino acid sequence of the phosphorylated 145-kD protein induced by H. pylori infection was identical to the H. pylori CagA sequence. These results reveal that the tyrosine-phosphorylated 145-kD protein is H. pylori CagA protein, which may be delivered from attached bacteria into the host cytoplasm. The identification of the tyrosine-phosphorylated protein will thus provide further insights into understanding the precise roles of CagA protein in H. pylori pathogenesis.

Outside-In signaling of soluble and solid-phase fibrinogen through integrin alphaIIbbeta3 is different and cooperative with each other in a megakaryoblastic leukemia cell line, CMK.

The function and the outside-in signaling pathways of alphaIIbbeta3 were examined in relation to cell adhesion using a megakaryoblastic leukemia cell line, CMK. After 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment, the cells adhered to the culture plate and underwent megakaryocytic differentiation with expression of alphaIIbbeta3. Binding of soluble fibrinogen to the cells via alphaIIbbeta3 was dependent on cell adhesion. Cell detaching reduced the affinity of this integrin for soluble fibrinogen, although its surface expression was almost unchanged. In contrast, detached cells became tightly adherent to the fibrinogen-coated plate (solid-phase fibrinogen). The same ligand, fibrinogen, present either in soluble or solid-phase form, triggered differential signaling pathways mediated by alphaIIbbeta3. By the stimulation with soluble fibrinogen, Syk was tyrosine-phosphorylated but FAK was dephosphorylated, whereas solid-phase fibrinogen promptly caused tyrosine phosphorylation of FAK followed by delayed phosphorylation of Syk. In addition, the binding of soluble fibrinogen to the cells adherent to fibrinogen-coated plate resulted in tyrosine phosphorylation of integrin beta3 and a complex formation of integrin beta3 with Syk. This implies the cooperation of both soluble and solid-phase fibrinogen-mediated signaling pathways.

Establishment of a new murine-phenotypic angiosarcoma cell line (ISOS-1).

A cell line, designated ISOS-1, was established from a tumor formed by transplantation of a human angiosarcoma into mice with severe combined immunodeficiency (SCID). The cells showed endothelial properties, based on the uptake of Dil-Ac-LDL and binding of UEA-I/GSA-I lectins, but were negative for CD11b and Pan Cytokeratin. However, the cells lost differentiated characteristics such as expression of von Willebrand factor, contact inhibition growth and tube formation activity. These findings indicate that ISOS-1 is a poorly-differentiated endothelial cell line. At the 81st passage, all of the cells were positive for H-2Dd in various intensity, but not HLA-ABC. The metaphase chromosomes consistently showed a characteristic mouse, but not human, telocentric form. Furthermore, this cell line produced fatal tumor growth in SCID mice and also in BALB/c mice. These results suggest that ISOS-1 is a murine-phenotypic angiosarcoma cell line.

Interleukin-3 activates Syk in a human myeloblastic leukemia cell line, AML193.

Protein-tyrosine kinases and phosphatases play an important role in cytokine-mediated cell growth. The proliferation of a human myeloid leukemia cell line, AML193, is dependent on interleukin-3 (IL-3) or granulocyte colony-stimulating factor. In the current study, we demonstrated that a non-receptor-type protein-tyrosine kinase, Syk, was immediately activated by the stimulation with IL-3 or granulocyte colony-stimulating factor in AML193 cells. We further investigated the relation of Syk with IL-3-mediated signaling and found that the IL-3 receptor beta subunit was immunoprecipitated with Syk. Since the IL-3 receptor beta subunit is known to mediate growth signaling, our results indicate that Syk may be involved in the proliferation of myeloid cells.

Cyclic AMP-elevating agents negatively regulate the activation of p72syk in N-formyl-methionyl-leucyl-phenylalanine receptor signaling.

Here we investigated the involvement of the non-receptor protein-tyrosine kinase p72syk in formyl methionyl-leucyl-phenylalanine (fMLP) receptor signaling. The activity of p72syk began to rise from 15 s and reached to maximum within 2-5 min after 5 microM fMLP stimulation in porcine polymorphonuclear neutrophils (PMNs). Cyclic AMP (cAMP)-elevating agents, prostaglandin E2 (PGE2) and forskolin, or dibutyryl cAMP partially suppressed p72syk activities stimulated by fMLP in PMNs. Pretreatment with an inhibitor of cAMP-dependent protein kinase abolished the suppression of the fMLP-induced p72syk activation by these cAMP-elevating agents. It was also observed that cAMP-dependent protein kinase phosphorylates p72syk on serine residues in vitro. These results indicate a possibility that cAMP-dependent protein kinase negatively regulates the activation of p72syk in fMLP-receptor signaling.

Translocation, activation and association of protein-tyrosine kinase (p72syk) with phosphatidylinositol 3-kinase are early events during platelet activation.

We have previously reported that a non-receptor-type protein-tyrosine kinase p72syk, exists in both membrane and cytosolic fractions in porcine platelets and is activated after thrombin stimulation. To facilitate the understanding of the function of p72syk, we have investigated the topological features, kinase activities and the interaction with another signal-transducing molecule, namely phosphatidylinositol 3-kinase, during platelet activation. Membrane and cytosolic fractions were separated from thrombin-treated porcine platelets, and the amount of p72syk was quantified by the immunoblot technique or the kinase activity of each fraction was determined by an immunoprecipitation kinase assay. After stimulation by thrombin, cytosolic p72syk rapidly translocated to the membrane fraction within 10 s and there was also a significant increase in the amount of p72syk in the cytoskeletal fraction. The autophosphorylation activity of membrane-associated p72syk significantly increased approximately tenfold and reached a maximum at 10 s; the activity subsequently decreased to almost the basal level within 120 s. For similar time courses, association of p72syk with phosphatidylinositol 3-kinase and tyrosine phosphorylation of p72syk were observed. These results suggest that translocation, activation, and association of p72syk with transducing molecules such as phosphatidylinositol 3-kinase, events which occur during platelet activation, may participate in early signal-transduction events.

CPTK71, a cytosolic protein-tyrosine kinase previously purified from bovine platelets, is identical with p72syk.

In the previous study, we reported purification and characterization of cytosolic protein-tyrosine kinase, CPTK71, from bovine platelets (Nakamura, S., Yanagi, S. and Yamamura, H. (1988) Eur. J. Biochem. 174, 471-477). In the present study, we have investigated the relationship between CPTK71 and p72syk which was purified from cytosolic fraction of porcine spleen and sequenced from porcine cDNA library. The elution patterns of CPTK71 activity in each purification step closely corresponded with the pattern of immunoreactivity with p72syk. Immunoprecipitation study revealed that specific antibodies against p72syk precipitated the activity of both phosphorylating [Val5]angiotensin II and autophosphorylation of highly purified CPTK71. In addition, the same result was obtained using another anti-p72syk antibody which recognized different amino acid sequence of 72syk. From these results, we conclude that CPTK71 is identical or closely related to p72syk.

Effects of DS-4574, a new orally active antiallergic agent, in experimental allergic and inflammatory models.

The anti-allergic and anti-inflammatory activities of DS-4574, which possesses leukotriene antagonism and inhibits the release of immunologically stimulated mediators such as histamine and leukotrienes, were evaluated in several animal models. DS-4574 had dose-dependent inhibitory effects on IgE-mediated passive cutaneous anaphylaxis and the passive Arthus reaction in rats and the phase I response of Forssman antibody-induced bronchoconstriction. In contrast, this compound had no effect on the phase II response of Forssman antibody-induced bronchoconstriction in guinea pigs, the reverse cutaneous anaphylaxis in rats, complement-dependent hemolysis of sheep erythrocytes and the delayed-type hypersensitivity induced by methylated bovine serum albumin in mice. The results obtained in a double sensitization with two IgE antibodies suggested that DS-4574, as well as disodium cromoglycate, did not impair antigen-antibody combination but prevents the release of chemical mediators such as histamine. DS-4574 also had a weak inhibitory activity on carrageenin paw edema in rats, arachidonic acid ear edema in mice and adjuvant arthritis in rats. In addition, this compound inhibited increased vascular permeability in rat skin induced by leukotriene D4 and platelet activating factor-induced pleurisy in rats in a dose-dependent manner. These results indicate that DS-4574 inhibited type III allergic reactions and some inflammatory reactions. Therefore, DS-4574 could be useful in the treatment of allergic diseases such as asthma.

Synthesis and antiallergy activity of [1,3,4]thiadiazolo[3,2-a]-1,2,3-triazolo[4,5-d]pyrimidin-9 (3H)-one derivatives. I.

A series of 6-substituted [1,3,4]thiadiazolo[3,2-a]-1,2,3-triazolo[4,5-d]pyrimidin-9(3H)-one derivatives 4a--z were synthesized from 5-substituted 1,3,4-thiadiazol-2-amines 5 by the following consecutive reactions: pyrimidine ring closure with bis(2,4,6-trichlorophenyl) malonate, nitration, chlorination, amination, hydrogenation and diazotization. The structure of 4 was confirmed by an alternate synthesis of 4, involving reaction of 5-substituted 2-azido-1,3,4-thiadiazole 13 with ethyl cyanoacetate, followed by the Dimroth rearrangement and ring closure. The antiallergic activities (anti-passive peritoneal anaphylaxis, anti-passive cutaneous anaphylaxis and anti-slow reacting substance of anaphylaxis activities) of the products were evaluated.

Antiallergic activity of 6-(2-cyclohexylethyl)-[1,3,4]thiadiazolo- [3,2-a]-1,2,3-triazolo-[4,5-d]pyrimidin-9(3H)-one (DS-4574) in rats.

The antiallergic activity of DS-4574 was evaluated in several commonly used rat models for allergic diseases. In passive cutaneous anaphylaxis, DS-4574 given intravenously and orally induced dose-dependent inhibition with ID50 values of 0.55 and 2.8 mg/kg, respectively. In contrast, this compound had no antagonistic activity against the histamine- and serotonin-induced cutaneous vascular permeability. In lung anaphylaxis, DS-4574 inhibited pulmonary function changes induced by the antigen in a dose-dependent manner when it was given intravenously and orally, the ID50 values being 0.04 and 0.89 mg/kg, respectively. DS-4574 also inhibited antigen-induced histamine and leukotriene release in passive peritoneal anaphylaxis following oral administration. In addition, this compound prevented antigen-induced histamine release in passively sensitized mast cells in vitro. These potent activities of DS-4574 in in vivo and in vitro models of immediate-type hypersensitivity reactions suggest that this compound could be useful in the treatment of allergic diseases including asthma.

Temporal minor slow and sharp activity in psychiatric patients.

TMSSA was found in 51 (4.7%) of 1091 psychiatric patients, with a higher incidence found in the older group (40 years of age or over). Although TMSSA was found in various disorders, a close association with cerebrovascular dementia was suggested. It was uncommon in patients with primary degenerative dementia. No increase of TMSSA was noted in cerebrovascular disorders without dementia. TMSSA was found fairly often in patients with affective disorders, neurosis, and other functional disorders, suggesting underlying mild cerebral dysfunction in some. TMSA, a variant of TMSSA, was very similar to TMSSA except for its greater frequency in cerebrovascular disorders without dementia.

[Reappearance and recovery of alpha rhythm after eye opening].

To obtain more informations from routine EEG recording, we studied changes in alpha rhythm after eye opening that had been almost disregarded hitherto. The study was carried out on 26 normal controls and 137 patients with four categories of diseases including 52 patients with psychiatric diseases, 8 with trifling neurological symptoms, 68 with defined organic brain diseases, and 9 with headache without defined organic cause. Following routine EEG recording, each subject was asked to keep their eyes opened for three minutes. The major changes in alpha rhythm after eye opening were composed of two patterns; "reappearance" and "recovery". The reappearance was defined as appearance of 5 or more alpha waves in series; and the recovery, as the amount and amplitude of the alpha rhythm became equal to those of the back ground alpha-rhythm which was seen while their eyes were kept closed. The reappearance was noticed in 84.6% of normal controls and in 89.1% of all patients. In patients with organic brain diseases, the reappearance was seen in 88.2%, while it was in 68.6% of the patients without organic brain disease. The recovery was noticed in 34.3% of all patients (p less than 0.01), while it was only in 15.4% of normal controls. The incidence of the recovery in the patients with organic brain diseases was 41.2%, the value of which was significantly higher than that of the rest of patients (28.8%, 15 out of 52 patients, p less than 0.05). These patients with the recovery tended to show slower frequency and higher amplitude in the back ground alpha-rhythm than the patients without recovery.

Mode couplings due to external forces distributed along a polarization-maintaining fiber: an evaluation.

A new interferometric scheme for measuring mode conversion distributed locally along a polarizationmaintaining fiber is presented. Using this technique the power coupling coefficient, varying with magnitude and angle of external pressure transversely applied to a fiber, was evaluated both theoretically and experimentally. The coupling point location is determined with +/- 1.5-cm accuracy and resolution of better than 10 cm for a 220-m long fiber having modal birefringence of 4.4 x 10 (- 4). The coupling coefficient was proportional to the external force in the range from 5 x 10 (- 3) to 0.1 kg/mm. The relationships determined experimentally reflected those predicted by theory.

Longitudinal coherence properties of light waves propagating through a birefringent fiber.

Longitudinal coherence properties of the waves propagating through a birefringent fiber are investigated theoretically and experimentally. Significant loss due to the polarization-dispersion slope is observed clearly for the interference between the two orthogonally polarized HE(11) modes. The results obtained experimentally reflect the theoretical predictions well for both the modulus of the degree of coherence and its curve shape versus the optical path difference in the wavelength region from 816 to 1540 nm.

Comparison of insulin-like growth factor I receptor and insulin receptor purified from human placental membranes.

Insulin-like growth factor (IGF)-I receptor purified from human placental membranes as previously described (LeBon, T. R., Jacobs, S., Cuatrecasas, P., Kathuria, S., and Fujita-Yamaguchi, Y. (1986) J. Biol. Chem. 261, 7685-7689) was characterized. The IGF-I receptor was similar to the insulin receptor with respect to subunit structure (beta-alpha-alpha-beta), apparent sizes of deglycosylated alpha (Mr = approximately 88,000) and beta (Mr = approximately 67,000) subunits, and amino acid composition of the subunits. Monoclonal antibody specific to each receptor recognized its own receptor whereas polyclonal anti-human insulin receptor antibody cross-reacted with the IGF-I receptor, indicating that the receptors share one or more antigenic sites. Further characterization of the purified IGF-I receptor tyrosine-protein kinase activity indicated that by analogy with the insulin receptor the monomeric alpha beta form of the IGF-I receptor appears to have higher kinase activity than the intact receptor in the alpha 2 beta 2 form. The most significant difference between the two receptors was found in the N-terminal amino acid sequences of their alpha subunits, which apparently show 60% identity. The IGF-I receptor alpha subunit lacks residues corresponding to the N-terminal 4 amino acids of the insulin receptor alpha subunit. These results provide the first direct proof that the IGF-I receptor is a molecule distinct from the insulin receptor despite numerous similarities.

Differential effects of aging on adrenocorticotropin receptors, adenosine 3'5'-monophosphate response, and corticosterone secretion in adrenocortical cells from Sprague-Dawley rats.

This study examined the effect of aging on adrenal cell function in Sprague-Dawley rats, as judged by the binding of iodinated Phe2, Nle4-ACTH-(1-38) to the adrenal cells, and the ability of the cells to respond to ACTH stimulation by the production of cAMP and corticosterone in vitro. Collagenase-dispersed adrenal cells obtained from 2-, 12-, and 18-month-old-rats were used. The maximum corticosterone concentration after incubation with ACTH was significantly lower (P less than 0.001) in the 12-month-old (40 +/- 7 ng/micrograms DNA) and 18-month-old rats (28 +/- 3 ng/micrograms DNA) compared to that in 2-month-old controls (102 +/- 9 ng/micrograms DNA). The ED50 values of ACTH-induced corticosterone production measured in the cell suspension were similar in 2- and 12-month-old groups (30 pg/ml), and the diminished production of corticosterone in the 12-and 18-month-old rats persisted after incubation with N6,O2-dibutyryl cAMP. Neither the number nor the binding affinity of adrenal receptors for [125I]I-Tyr23,Phe2,Nle4-ACTH-(1-38) changed from 2-12 months of age. Furthermore, increases in concentrations of intra- and extracellular cAMP after ACTH stimulation were not significantly different in the 2-, 12-, and 18-month-old groups. Similarly, adrenal hydrolysis of cAMP by low and high Km phosphodiesterases did not change significantly with advancing age. These results provide strong evidence that there is a diminished capacity for corticosterone production with aging in the rat, and that the site of the defect lies distal to binding of trophic hormone to its receptor and to the production of its secondary messenger. Finally, an age-related decline in adrenal steroidogenic capacity could be viewed as a counterregulatory mechanism invoked in old rats to compensate, at least partially, for elevated plasma ACTH and corticosterone concentrations.