241 research outputs found
変形性膝関節炎患者膝関節液中の細胞外マトリックス成分と可溶性VCAM-1
It is generally accepted that the number of patients with osteoarthritis (OA) of the knee must increase in the future because of the increase of the aged. The pathogenic mechanisms of OA haven't been elucidated yet, but it is supposed that mediate pannus may cause the factors for irreversible cartilage damage in the inflammatory process. Therefore it is very important to find and predict OA at the early stage. Now there were many indices to determine joint damage with OA, for example, cytokines including IL, TNF, TGF, matrix metalloproteinase (MMPs), tissue inhibitor of metalloproteinase (TIMPs), proteoglycan (PG), hyaluronate (HA), super oxide, adhesion molecules including, ICAM-1, VCAM-1 and PECAM. In this study, we measured that the soluble VCAM-1 (sVCAM-1), soluble collagen (sCOLL), heparansulfate proteoglycan (HSPG), uronic acis and CRP in synovial fluids (SF) with OA. A positive correlation between the level of sVCAM-1 and sCOLL (ρ = 0.652) was noted. However there was no factor showing a correlation with CRP. It was suggested that the levels of sVCAM-1 and sCOLL in the synovia fluid were active inflammation indices of knee damage, as the different aspects from CRP. The level of HSPG also showed a unique trend because HSPG on the cell surface was quickly digested endocytosis, so that it was not suitable for the inflammatory index in synovial fluid with OA.変形性膝関節炎(OA)患者は,高齢化社会の到来とともに今後ますます増加するものと考えられる。OAの発症メカニズムについては,関節軟骨の破壊・修復やパンヌス形成に関わる多くの関連因子(サイトカイン,マトリックス分解酵素,分解酵素抑制因子,プロテオグリカン,活性酸素,細胞接着因子など)について,多面的に研究が進んでいる。しかし,膝関節の炎症初期の病態のメカニズムについては,なお詳細に解明されておらず,関節炎初期の炎症指標の検索が今後さらに必要と考えられる。本研究で私たちは, OA患者膝関節液中の細胞外マトリックス構成成分であるコラーゲン,へパラン硫酸プロテオグリカン,ウロン酸および炎症の最も初期に血管内皮細胞に出現するVCAM-1等の可溶化成分濃度を測定した。患者膝関節液(SF)中の可溶性コラーゲン量と可溶性VCAM-1(sVCAM-1)量とは有意な正の相関関係が(ρ=0.652)観察された。しかし,ヘパラン硫酸プロテオグリカン(HSPG)と可溶性コラーゲン(sCOLL)値間には有意な相関関係は確認されなかった。また,一般に炎症の指標として用いられるCRP値とも比戟検討したが,CRP値と他の測定項目値とは一定の相関関係は存在しなかった。以上の結果より,OA患者のSF中の炎症マーカ-として,急性炎症時には従来の炎症指標蛋白質であるCRP値が有力であるというSipe JDの報告から推測するとsCOLLとsVCAM-1はCRP値と一定の相関を示さないことより,CRPの動態とは異なった炎症指標としての有用性が考えられる。しかし,HSPGはエンドサイトシスにより速やかに代謝されるためSF中への解離が少なく,炎症指標としての可能性は低いと考えられる
リウマチ因子陽性ヒト血清中の免疫グロブリンGクラスリウマチ因子と免疫複合体ならびにCRPとの相関性
We measured the concentration of three immunoglobulin classes of rheumatoid factors (Ig-RFs), C3d binding IgG immune complex (C3d-IC), C1q binding IgG immune complex (C1q-IC) and C-reactive protein (CRP) in 74 samples of human sera with high levels of RF (24.0~2350.0IU/ml). In sera with high levels of C3d-IC (>15.0μg/ml), there was a positive correlation between the levels of CRP and the IgG-RF, but there was no correlation between the levels of CRP and the immune complexes (C3d-IC and C1q-IC). And then, there was a positive correlation between the levels of CRP and IgG-RF or C3d-IC and IgG-RF when the levels of C1q-IC in patients sera were higher than 80.0μg/ml. However, there was no correlation between the levels of CRP and C1q-IC in these patients sera containing high levels of both C3d-IC and C1q-IC. These results indicated that the determination of C3d-IC, C1q-IC, IgG-RF and CRP in human sera containing RF denote different implications as inflammatory indexes on progression of rheumatoid arthritis and other autoimmune diseases.一般に慢性関節リウマチの診断ならびに活動性の一指標として、日常臨床検査では患者血清中のリウマチ因子の測定が実施されている。本論文ではRF検査依頼が有り、高RF値(24.0~2350.0IU/ml)を示した患者血清74検体について、免疫グロブリンクラス別RF値を測定するとともに自己IgGと補体との免疫複合体(IC)であるC1q-IgG・IC(C1q-IC)、C3d-IgG・IC(C3d-IC)ならびにCRP値を測定し、それらの測定値間の相関性について統計学的検討を行った。C3d-IC値が15.0μg/ml以上の高値を示す患者血清では、CRP値とIgGクラスのRF(IgG-RF)値間については正の相関結果がが得られた。また、C1q-IC値が80.0μg/ml以上の場合にもCRP値とIgG-RF値間およびC3d-IC値とIgG-RF値間では正の相関結果が得られた。しかし、C3d-ICならびにC1q-ICともに高値例の患者血清中のCRP値とC1q-IC値間には有意な相関は観察されなかった。以上の結果より、慢性関節リウマチの活動度を判定する上で、従来から炎症マーカーとしてCRP値が利用されているが、血清中に免疫複合体が高レベルに検出される患者については、炎症の指標としてCRP以外にもC1q-ICやC3d-ICおよびIgG-RF等を加えた総合的な判断が必要であることが示唆される
長期ホルマリン固定により失活したProliferating Cell Nuclear Antigen (PCNA) の免疫反応性回復条件の基礎的検討 ―マイクロウェーブ、オートクレーブの影響について―
Using paraffin-embedded tissue sections of liver cancer obtained from autopsy which had been preserved in 10% buffered formalin solution for 6 months while PCNA immunoreactivity was lost, we examined the effects of heat processing by either microwave(MW) and autoclave(AC) in the presence of various processing solution. It appeared that AC processing took shorter time period than MW irradiation to restore equal immunoreactivity. With regard to immunoreactivity retrieval by MW irradiation,however, variation of the degree of retrieval depending on processing time was smaller than in AC, and so the stable consequences were obtained. Although AC processed tissues tended to be stained deep, prolonged processing time presented strong background staining and blurred nuclear margins which made it difficult to estimate the positive cell count. As for the effects of processing solution, there was little difference in retrieval of PCNA among 0.01 M citrate buffer (pH 6.0), saturated solution of lead thiocyanate and distilled water, but the least background staining was observed with distilled water. These observations suggest that MW irradiation of which effect of retrieval is less dependent of processing time and with the least background stainability, is superior to AC processing for PCNA immunoreactivity retrieval on formalin-fixed tissues.10%緩衝ホルマリンに6ヶ月間浸漬していた剖検材料(肝臓癌)のパラフィン包埋後の組織切片を用いて、ホルマリンの固定作用により失活したPCNAの免疫反応性の回復にマイクロウェーブ(MW)及びオートクレーブ(AC)による熱処理とその時用いる処理溶液が、どのような影響を与えるかについて検討した。その結果、同等の免疫反応性を回復するのには、AC処理の方がMW照射より短時間でよいことが解った。しかし、MW照射による免疫反応性回復では、処理時間による影響がAC処理に比べて少なく、安定した結果が得られた。また、PCNAの染色所見については、AC処理の方が濃く染まる傾向が見られた。しかし、処理時間が長くなるとバックグラウンドの染色性が高くなる、核の周囲ににじみ現象が見られる等の所見があり判定に困難をきたした。一方、処理溶液についてはクエン酸(0.01M pH6.0)、チオシアン酸鉛飽和溶液、蒸留水について検討を行った結果、PCNAの回復には差は認められなかったが、バックグラウンドの染色については蒸留水が最も少なかった。以上の結果により、ホルマリン固定により失活したPCNAの免疫反応性を回復するには、MW照射の方がAC処理より処理時間に関係なく安定した染色性が得られ、なおかつバックグラウンドの染色性が少ない等の点で優れていることが示唆された
Hydrodynamic stress stimulates growth of cell clusters via the ANXA1/PI3K/AKT axis in colorectal cancer
Cancer cells are exposed to various stresses in vivo, including hydrodynamic stress (HDS). HDS on cancer cells in the blood stream can influence the metastatic potential. Recent studies revealed that circulating tumor cell clusters are more responsible for metastasis than circulating single cells. Nevertheless, most studies on HDS are based on single cells prepared from established cancer cell lines. Here, we used cancer tissue-originated spheroids (CTOS) as a patient-derived, 3D organoid model to investigate the effect of HDS on cancer cell clusters. We found that HDS induced the growth of cancer cell clusters in a population of colorectal CTOSs. Microarray analyses revealed that the multifunctional protein, Annexin 1 (ANXA1), was upregulated upon HDS exposure. Chemically-induced membrane damage also triggered the expression of ANXA1. A knockdown of ANXA1 revealed that ANXA1 regulated HDS-stimulated growth in colorectal CTOSs. Mechanistically, activating the PI3K/AKT pathway downstream of ANXA1 contributed to the phenotype. These findings demonstrate that HDS induces the growth of cancer cell clusters via ANXA1/PI3K/AKT axis, which helps to elucidate the pro-metastatic feature of circulating cancer cell clusters
Mechanism of Phospholipase C Activation through the T Cell Receptor/CD3 Complex - Evidence for the Activation without Involvement of a Guanine Nucleotide-binding Protein -
ヒト血清リポタンパク質とシクロデキストリンのアガロースゲル内における相互作用
Cyclodextrins (CDs) are cyclic oligosaccharides with low molecular weight. They have been known to bind to some serum lipoproteins and to form complexes. To elucidate whether each serum lipoprotein subclass could be separated by electrophoresis using CDs, we performed preliminary experiment, in which lipoprotein-CDs interaction was examined on electrophoresis with agarose gel. When the supporting agarose gel containing both α-CD and β-CD was prepared and was applied to isoelectric focusing for fractionating serum lipoproteins, apoB lipoproteins were found to be clearly separated into several fractions on this electrophoresis. This finding suggested that apoB lipoprotein may be detected as isolated form by arranging amounts of added CDs.シクロデキストリンは低分子量の環状オリゴ糖で, リポタンパク質と複合体を形成する。シクロデキストリンを使用した電気泳動による血清リポタンパク質サブクラスの分離の可能性を明らかにするために, リポタンパク質とシクロデキストリンとの泳動用ゲル内での相互作用について検討をおこなった。その結果,α-CDとβ-CDの2種類のCDをアガロースゲルに添加し,このゲルを泳動用支持体として等電点電気泳動をおこなったところ,apoB含有リポタンパク質が数分画分離された。したがって,ゲル内へのシクロデキストリンの適正な添加条件が設定できるならば,apoB含有リポタンパク質サブクラスの分離が可能となることが示唆された
Differential responses to high- and low-dose ultraviolet-B stress in tobacco Bright Yellow-2 cells
Difference in the Ca2+ Responses of Various Human T Cell Lines to Stimulation of the T Cell Receptor/CD3 Complex with Monoclonal Antibodies
IgG Heavy-Chain Subclass Typing of Human Serum M-Protein -Optimal condition for the separation and the detection with high resolution of agarose gel isoelectrofocusing-
IgG Heavy-Chain subclass typing of human serum M-protein were isoelectrofocussed (IEF) in agarose gel, and then the bands of IgG were detected with peroxidase-conjugated anti-human IgG1-4 monoclonal antibodies on the same isoelectrofocussed agarose gel plate. This IEF enzyme immunodetection patterns were composed of four to eight discrete bands (The range of pI was 6.0 to 9.0). These bands were dependent on immunofixationbands. It was very specific and clear to detect the subclass of IgG Type M-protein in human serum using this method
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