今回著者は歯周疾患の免疫組織化学的研究の一環として, イヌ歯肉コンポーネントに対するモノクローナル抗体を作製し, その対応抗原の組織局在性および免疫学的性状について検索した。
3回, 17日間の免疫ににり感作されたマウス脾細胞とマウス骨髄腫細胞P3U1との細胞融合により, 数種類の興味ある抗体産生性ハイブリドーマを得, 限界希釈法によるクローニングでD1, D2と命名する2種類のモノクローナル抗体を得た。
1, D2ともIgMで, D1はD 歯肉上皮の細胞膜および毛細血管, D2は細胞質および毛細血管と反応した。また自腔内 (舌, 硬口蓋, 口唇, 頬粘膜) および皮膚の重層扁平上皮においても, 歯肉に対するそれと同様な染色パターンを示し, 重層扁平上皮以外でも胃, 小腸, 肝臓, 腎臓腹大動脈など広範囲において興味ある分布状況を示した。
さらに生化学的検索により, モノクロ抗体D1の対応抗原の分子量は50Kダルトンの分子であることが判明した。
以上のことからイヌ歯肉に対するモノクロ抗体の作製が可能であり, その特異性を利用した研究は今後歯周疾患を免疫組織学的に研究する上で, 有用な手段となることが明らかとなった。

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Cortical motor areas of the jaw and orofacial regions were studied by intracortical micro-stimulation (ICMS:<30μA, 200μsec duration and 300 Hz in 40 msec train duration) in the cat lightly anesthetized with Ketamine-HCL. EMG activities produced by ICMS to the cerebral cortex were bi-polarly recorded from jaw and orofacial muscles, and the onset of latencies of EMG activities were analyzed. Furthermore, patterns of motor effects were examined by the ablation of jaw and orofacial motor areas. Jaw and orofacial motor effects were produced by ICMS delivered to anterior parts of the coronal and lateral sigmoid gyri (C area), and to the middle to the anterior part of the orbital gyrus (O area). Cytoarchitectonically, jaw and orofacial motor areas were restricted to areas 3a, 4 γ and6aβ in the C area, and to areas 43 and 6aβ in the O area. Different patterns of movements in jaw and orofacial regions were evoked from these two areas. In the C area one or, at most two, types of simple movements were produced by ICMS to one location, while in the O area more co-ordinated movements than those in the C area were produced and rhythmic jaw movement was evoked by repetitive stim-ulation (<30μA, 30 Hz and 200μsec duration) to the O area. The onset of latencies of the EMG activities of the facial muscles of the whisker pad and eyelid regions, evoked by ICMS to the O area were longer than those evoked by ICMS to the C area.

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Light and electron microscopic observations were carried out on the secretory granules of the labial gland of the lizard, Japalura polygonata polygonata. The results obtained are.
1) Japalura polygonata polygonata has superior and inferior labial glands, which can be described as a branched tubular type.
2) The glandular cells are arranged in a layer and appear to be columnar in shape.
3) The secretory granules of the superior labial gland stain intensely with naphthol yellow S and iron hematoxylin and are weakly positive with PAS and alcian blue.
4) The secretory granules of the superior labial gland can be divided into two types when examined with the electron microscope. One is of moderate electron density, while the other has a moderate dense core and a narrow less dense halo. The PA-methenamine silver method shows a positive reaction which is limited to the narrow halo of the latter granule.
5) The secretory granules of the inferior labial gland except those of the acini which are oriented superolaterally in the lower lip stain intensely with PAS and alcian blue, and they are also slightly positive with naphthol yellow S and iron hematoxylin.
6) The secretory granules of the inferior labial gland can be classified into six types by the degree of the electron density of their core and halo. One of these types has a tripartite structure. Almost all of the secretory granules are positive for the PA-methenamine silver reaction.
These findings suggest that the superior and inferior labial glands of Japalura polygonata polygonata have different characteristics from each other, because the secretory granules of the superior labial gland mainly contain protein, while those of the inferior labial gland contain mucopolysaccharides.

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The adrenergic nerve terminals in the rat submandibular gland, with the conventional method of fixation, contained many small round clear or granular vesicles; whereas, with the chromaffin reaction method of Tranzer and Richards, frequent tubular or flattened vesicles were observed in addition to the small round granular or clear vesicles. The factors influencing the occurrence of tubular or flattened vesicles after the chromaffin reaction method were examined.
The fixation procedure of chromaffin rection method was divided into three steps; prefixation, storage in the reaction medium and postfixation. Total of 11 experimental series were set up replacing some of these steps with corresponding ones used in conventional fixation method. The tubular or flattened vesicles were exclusively observed in the experimental series using 0.2M chromate buffer as the storage medium. With 0.08M chromate buffer which has the same osmolarity with 0.1M cacodylate buffer used in conventional fixation method, tubular or flattened vesicles could not be observed. On the other hand, the variation of the pH (from 6.0 to 7.2) of the buffer did not alter the profiles of synaptic vesicles; whereas, many synaptic vesicles were stained with dense chromaffin-reaction-products at pH 6.0 but not at pH 7.2.
In conclusion, the tubular or flattened vesicles may be an artefact resulting from high osmolarity of the storage buffer used in the chromaffin reaction method.

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Adherence of a Gram positive, anaerobic bacterium Propionibacterium ances to epithelial cells was examined using an in vitro assay system. The adherence of three strains ATCC 6919, 11827, and 11828 of P.acnes to human buccal epithelial cells was dependent on bacterial concentration, incubation time, and pH. Heat treatment of the bacterial cells decreased the subsequent bacterial adherence to the epithelial surfaces but the bacterial adherence was not completely eliminated by heat treatment. Trypsin added to an adherence assay system significantly inhibited the adherence of P. acnes ATCC 11828.
Scanning electron microscopy demonstrated projecting structures up to 300mμ long on the surface of strain ATCC 11828 associated with the bacterial attachment to the surface of the epithelial cells. Negative staining technique and an ultrathin section of a strain ATCC 11828 adherent to an epithelial cell also showed fuzzy structures mediating the bacterial binding to the epithelial cell. These results suggest that P.acnes adheres to human buccal epithelial cells by the heat-resistant fuzzy strurtures on the surface of the bacteria.

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あらかじめ⁴⁵Caをラベルしたラット胎児骨の培養法を用いて, 種々のサイトカイン活性溶液, 精製サイトカインあるいは遣伝子導入大腸菌由来のレコンビナントサイトカインについて骨吸収刺激活性を測定した。その結果, PHA・Pで刺激したヒト末梢血単核球の培養上清やマウス腹腔マクロファージやウサギ炎症性多形核白血球由来の部分精製IL-1様活性溶液に骨吸収刺激活性が認められた。これらの成績を下にして, 種々の精製サイトカイン, あるいはレコンビナントサイトカインの骨吸収活性を調べたところ, ヒトレコンビナントIL-1 (β) とレコンビナントTNFの両マクロファージ因子に骨吸収刺激活性が認められた。T細胞由来のサイトカインではレコンビナントIL-2, CSF (G), IFNγ には骨吸収活性が見出されなかったが, マウスWEHI-3細胞から精製したIL-3溶液に活性が認められた。以上の成績は, 骨吸収機構にはマクロファージや多形核白血球の関与が明らかなこと, そして, 骨髄細胞の分化が骨吸収 (そして恐らく骨形成) 機構に深く関与していることを示唆するものと考えられる。

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Trichomonas tenax (T. tenax) が合成組織培養培地で培養できるかおよび随伴細菌に対する抗体と補体の組み合わせ処理でその随伴細菌が除去できるか否かを調べた。RPMI1640およびイーグルMEM培養液はそれぞれ単独でも, T. tenaxをいくらか増殖でき, かつ血清を添加すると, 濃度に比例してその増殖は増加した。また, 抗体と補体の組み合わせ処理で, 48時間培養まで, その随伴細菌の増殖がほぼ完全に抑制された。T. tenaxの純培養には成功しなかったが, その培養が, 従来の非合成のEgg-York培地だけではなく, 合成組織培養培地でも培養できることまた抗体存在下で随伴細菌の増殖を抑制できることは, 抗生物質処理のような耐性獲得がないことから, それぞれ, T. tenaxの生物学的性状の研究に役立つと思われる。また, 本実験に用いた組織培養培地で培養細胞とT. tenaxのいずれも培養できることから, 培養細胞とT. tenaxとの相互関係の研究にも利用できる。

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低亜鉛食飼摂取ニワトリの舌背表面を走査電子顕微鏡で観察すると, 正常のニワトリでは, 舌背前2/3の部位に密に分布する昆布状の糸状乳頭の分布頻度が著しく減少していた。正常のニワトリでも, 低亜鉛食飼摂取ニワトリでも, 舌背後1/3の部位は, ほぼ同様の表面形態を呈しており, 糸状乳頭は分布しなかった。走査電子顕微鏡による強拡大像においては, 正常ニワトリの舌背後1/3の部位の粘膜上皮表面には, やや不明瞭なmicroridgeあるいはmicropitとよばれる小孔状構造が観察されるのに対して, 低亜鉛食飼摂取のニワトリの舌背後1/3の部位の粘膜上皮表面には, 輪郭の明瞭なmicroridgeが広く観察された。透過電子顕微鏡で観察すると, 舌背前2/3の部位の糸状乳頭直下の角化層の厚みは, 低亜鉛食飼摂取群において著明な減少をきたしており, 比較的多数の核が角化層に相当する部位でも観察されるようになる。舌背後1/3の部位の粘膜上皮では, 正常なニワトリで観察できる表層直下の高電子密度の薄い細胞層が, 低亜鉛食飼摂取ニワトリでは, ほとんど認められず, 表層にいたるまで隣接細胞間には, 多数のdesrnosomeが介在している。

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