Masao Aono

Effects of polymyxin B on superoxide anion release and priming in human polymorphonuclear leukocytes.

We studied the effect of a potent inhibitor of protein kinase C, polymyxin B (PMXB), on superoxide anion (O−2 release by human polymorphonuclear leukocytes (PMNL). PMXB was compared with another inhibitor of protein kinase C, 1‐(5‐isoquinoline‐sulfonyl)‐2‐methyl piperazine (H‐7). Both PMXB and H‐7 inhibited phorbol myristate acetate (PMA)‐stimulated O−2 release. Formyl‐methionyl‐leucyl‐phenylalanine (FMLP)‐stimulated O−2 release by cytochalasin B–treated PMNL was not inhibited significantly by either PMXB or H‐7. 1‐Oleoyl‐2‐acetyl‐glycerol (OAG,25–100 μM) stimulated PMNL to release O−2 with a long lag‐time (8–10 min). Although H‐7 inhibited OAG‐stimulated O−2 release, PMXB augmented the OAG‐stimulated response by increasing rate and reducing lag time. The augmenting effect of PMXB was evident only when added after stimulation by OAG, with maximum effect observed at 3 min after addition of OAG. The augmenting effect was also seen with PMXB immobilized on agarose beads. PMXB did not affect the respiratory burst response to 1,2‐dioctanoylglycerol. PMXB‐augmented, OAG‐stimulated O−2 release was inhibited by the addition of H‐7 before OAG. In contrast to the effect on O−2 release, OAG‐stimulated protein phosphorylation was inhibited similarly by either PMXB or H‐7, when these agents were added 3 min after stimulation by OAG. These results suggested that initial activation of protein kinase C by OAG is essential for O−2 release, but that PMXB acts in a manner independent from protein kinase C to augment OAG‐stimulated O−2 release. When priming by OAG for enhanced O−2 release (as opposed to direct stimulation of O−2 release) in FMLP‐stimulated PMNL was examined, PMXB inhibited O−2 release in OAG‐primed PMNL, suggesting that protein kinase C is involved in priming of PMNL by OAG.

The Modulation of Polymorphonuclear Leukocyte Function by Bacteroides Gingivalis

B acteroides gingivalis has been recognized as a pathogen causing adult periodontal disease. In this study, we examined the culture supernatants of B. gingival is for their ability to alter the function of polymorphonuclear leukocytes (PMNs). After a 60-minute incubation with the culture supernatants of B. gingivalis, it was found that there was a significant reduction of f-methionyl-leucyl-phenylalanine (FMLP) receptor in human and guinea pig PMNs. Superoxide anion (O- 2) production of PMNs stimulated by FMLP was also suppressed when cells were pre-incubated with the culture supernatants of B. gingivalis. A similar suppression was observed when these cells were stimulated by phorbol myristate acetate. An exohemagglutinin fraction of the culture supernatants showed suppressive activity at a low dose (10 μg/mL). The data presented in this paper suggest that B. gingivalis releases substances which suppress the functions of PMNs via the modulation of cell surface receptors and internal cellular events. These suppressive effects may be important in the pathogenesis of this micro-organism.

Effects of disodium ethane-1-hydroxy-1, 1-diphosphonate (EHDP) on interleukin 1 production by macrophages

The effects of disodium ethane‐1‐hydroxy‐1,1‐diphosphonate (EHDP) on the in vitro functions of guinea pig macrophages were studied. A high dose (1 mg/ml) of EHDP inhibited interleukin 1 (IL 1) production by oil‐induced peritoneal macrophages stimulated with muramyl dipeptide (MDP), lipopolysaccharide (LPS), phorbol myristic acetate (PMA), heat‐aggregated IgG2 or calcium ionophore A23187. On the other hand, low doses (<0.125 mg/ml) of EHDP augmented the MDP induced IL 1 production by macrophages. This biphasic effect was also observed when macrophages were exposed to EHDP at 37 C for 24 hr and then stimulated with IL 1 inducers. Superoxide anion generation induced by formyl peptide or PMA was not affected by preincubation of the macrophages with doses of EHDP up to 1 mg/ml. Adherence and spreading of macrophages was inhibited by EHDP in a dose dependent manner without affecting cell viability. These results demonstrated that EHDP acted on macrophages directly and modulated IL 1 production in vitro.

Long-term occlusal guidance of a severely intoxicated patient with yusho (PCB poisoning): a case report.

The peculiar dentoorofacial characteristics and 12 years of interdisciplinary management of a patient who was severely intoxicated with the man-made chemical polychlorinated biphenyls (PCBs) are described in this case report. Cephalometric measurements showed that the SNA and SNB angles were reduced but that the developments in height and skeletal maturity were in normal ranges. Gingival hyperpigmentation that was caused by high blood PCB concentration was extremely slow to fade. A cystic mass, diffused trabeculae, and irregular calcification, which were shown on the radiograph and which were caused by high blood levels of PCB, changed gradually. However, the patient had periodontal disease because of horizontal alveolar bone resorption and a deep periodontal pocket, despite good plaque control. After the PCB poisoning the tooth roots were hypoplastic and dilacerated. One root was extracted because of chronic periodontitis. Some teeth were impacted, malposed, or ankylosed.

Muramyl Dipeptide Induced Augmentation of the Proliferative Response of Thymocytes to Phytohemagglutinin

N‐Acetylmuramyl‐l‐alanyl‐d‐isoglutamine (MDP) augmented the proliferative response of thymocytes to phytohemagglutinin (PHA). The augmenting effect of MDP disappeared by passage of glass‐nonadherent thymocytes through Sephadex G‐10 (G‐10) column or by removal of low density cells by the Ficoll‐Conray gradient centrifugation. The diminished augmenting effects of MDP on the proliferative response of glass‐nonadherent‐G‐10 nonadherent thymocytes was restored by the addition of the G‐10 adherent cells. When G‐10 adherent cell fraction was extensively depleted of macrophages by glass adherence and EA‐rosetting, it was found that neither the macrophage‐depleted G‐10 adherent cell fraction nor the macrophage fraction supported by itself the proliferative response of G‐10 nonadherent thymocytes. However, addition of macrophage fraction together with the macrophage‐depleted G‐10 adherent cells did support the proliferation of G‐10 nonadherent thymocytes. It was further shown that peritoneal exudate macrophages could be substituted for thymic macrophage fraction. These results suggested that both the G‐10 adherent‐glass nonadherent cells and macrophages were essential for the MDP‐induced augmentation of the proliferative response of thymocytes to PHA and these cells exerted different accessory roles in this response.

[Basic studies on CaO-P2O5-MgO-SiO2-CaF system glass ceramics. 1. Morphology under the phase-contrast microscope and growth of cultured cells].

In order to determine the biocompatibility of glass ceramics which is one of the new biomaterials, in vitro studies were carried out by a cell culture method using four established cell lines. Materials used were glass ceramic disks with a diameter of 3 mm, and polystyrene coverslips of the same size as controls of the growth curve. Cells of each line were inoculated into 24-well multiplates at an appropriate density onto glass ceramic disks, and examined by phase contrast microscopy on the 1st, 3rd, 6th and 8th day. In addition, doubling time and saturation density were calculated from the growth curve. The results obtained were as follows. 1) Phase-contrast microscopy revealed that cells of each line attached to the disk within 24 hours and their numbers increased with time. After 8 days of cultivation, all of them reached confluence. 2) Contact with the glass ceramics did not cause cellular death or degeneration. Furthermore, the cultured cells showed the same morphological features as the control cells. 3) According to the growth curves, doubling time of all cells cultured with glass ceramics was shorter than that of the control cultures. On the other hand, saturation density was reduced to a minimum of 80% of the controls. These findings led to the conclusion that glass ceramic materials do not prevent the growth of cultured cells. According to the above results, glass ceramics possess the characteristics needed for bone grafts and implant materials.

Immunohistological study on movement of immune cells in periodontitis. 3. Analysis of T cell subsets.

辺縁性歯周炎歯肉組織中において, T cellおよびそのサブセットとランゲルハンス細胞 (LC) の動態を中心に, 免疫担当細胞についてモノクローナル抗体を用いた免疫組織化学的検索を行った。対照には歯肉炎罹患歯肉を用いた。その結果, 総浸潤細胞数の増加につれ, 浸潤細胞中のTcel1の数的比率は低下し, Bcell, マクロファー一ジ, および上皮細胞中のLCは増加する傾向にあった. T suppressor/cytotoxic cell (Ts/c) の比率は細胞浸潤 (+), (++) の場合共に対照群に比べて有意に低下し, このためT helper/inducer cellとTs/cの比は対照群に比べて大きくなっていた。また, 上皮細胞に対するLCの数的比率の増加と, Tcellおよびその各サブセットのそれが相関しており, 両者の関連の深さが伺われた。

[Bactericidal effect of human polymorphonuclear leukocytes--superoxide anion production of human polymorphonuclear leukocytes from peripheral blood, or gingival crevicular fluid].

辺縁性歯周炎発症における多形核白血球 (PMN) の役割を調べるために, 末梢血及び歯肉溝i浸出PMNの歯周病原性細菌に対する活性酸素 (とくにスーパーオキサイド: O2-) の産生やミエロペルオキシダーゼ (MPO) の測定を行い以下の所見を得た。1. 健康ヒト末梢血, 歯周炎患者末梢血及び歯肉溝滲出液中のPMNは, いずれも各種嫌気性菌に対してO2-産生を示した。2. 各種嫌気性菌の比較では, Actinobacillus actinomycetemcomitans, Bacteroides gingivalis及びcapnocytophagaに対してO2-産生が強く認められた。3. Bacteroides gingivalis菌体に対するPMNからのO2-産生は, 本菌が生菌の場合でも死菌の場合でも同程度に認められたが, 80℃ 以上の熱処理を加えるとO2-産生の低下が観察された。4. PMNのMPO活性については, 歯肉溝浸出液由来のPMNの方が末梢血よりも低下している症例が見られた。

Immunohistological study on movement of immune cells in periodontitis. 2. Movement of langerhans cells in periodontitis gingiva.

辺縁性歯周炎の発症年代と炎症の程度によって, ランゲルハンス細胞 (LC) の動態に相違があるかどうかについて検索を行った。慢性辺縁性歯周炎罹患歯肉52症例, 78部位について, ウサギ抗ヒトS-100蛋白を一次抗体としたペロキシダーゼ抗-ペロキシダーゼ (PAP) 法を用いて, 歯肉上皮内のLCを同定し, 上皮細胞中に占めるその割合を算出した。対照には臨床的健康歯肉および歯肉炎罹患歯肉を用いた。その結果, LCは炎症性細胞浸潤が多くなる程増加する傾向があった。辺縁性歯周炎と対照群とでは, 高年層の場合に限り, 前者に有意に多く, 若年層と対照群とでは有意差はなかった。これらのことからLCは炎症歯肉において抗原提示細胞としての機能を発揮しているものと考えられ, さらに若年層の辺縁性歯周炎は中高年層のそれと病態が若干異なる可能性が示唆された。

Epidemiological study of periodontal disease in Fukuoka region. Report 2 Prevalence of periodontal disease in th middle and old age people.

一般の中高年齢者107名を対象として歯周疾患の疫学調査を行った。診査は, 残存歯数, modified-Periodontal Disease Index (m-P.D.I.) 歯肉出血率, 治療必要度, 歯石沈着の有無について行い, アンケートとして口腔清掃指導を受けた経験の有無を調べた。その結果, 残存歯数及び歯石沈着の見られる者の割合の増齢的な減少が認められた。m-P.D.I. の年代別平均値は増齢的に増加しており, 特に深い歯周ポケットを有する者の割合が増加していた。また増齢的に, 外科的治療を含む複雑な治療を必要とする者の割合も増加しており, 増齢的な歯周疾患の悪化傾向が認められた。歯肉出血率は50代が他の2者に比べて有意に低く, 年齢による歯周疾患の病態の違いが示唆された。口腔清掃指導を受けた経験を有する者は全体の24.3%で, 経験の有無による口腔内の状態に差は認められなかった。