Multiple functions of the von Willebrand Factor A domain in matrilins: secretion, assembly, and proteolysis

The von Willebrand Factor A (vWF A) domain is one of the most widely distributed structural modules in cell-matrix adhesive molecules such as intergrins and extracellular matrix proteins. Mutations in the vWF A domain of matrilin-3 cause multiple epiphyseal dysplasia (MED), however the pathological mechanism remains to be determined. Previously we showed that the vWF A domain in matrilin-1 mediates formation of a filamentous matrix network through metal-ion dependent adhesion sites in the domain. Here we show two new functions of the vWF A domain in cartilage-specific matrilins (1 and 3). First, vWF A domain regulates oligomerization of matrilins. Insertion of a vWF A domain into matrilin-3 converts the formation of a mixture of matrilin-3 tetramer, trimer, and dimer into a tetramer only, while deletion of a vWF A domain from matrilin-1 converts the formation of the native matrilin-1 trimer into a mixture of trimer and dimer. Second, the vWF A domain protects matrilin-1 from proteolysis. We identified a latent proteolytic site next to the vWF A2 domain in matrilin-1, which is sensitive to the inhibitors of matrix proteases. Deletion of the abutting vWF A domain results in degradation of matrilin-1, presumably by exposing the adjacent proteolytic site. In addition, we also confirmed the vWF A domain is vital for the secretion of matrilin-3. Secretion of the mutant matrilin-3 harbouring a point mutation within the vWF A domain, as occurred in MED patients, is markedly reduced and delayed, resulting from intracellular retention of the mutant matrilin-3. Taken together, our data suggest that different mutations/deletions of the vWF A domain in matrilins may lead to distinct pathological mechanisms due to the multiple functions of the vWF A domain

Clinical outcome of arthroscopic capsular release for frozen shoulder: essential technical points in 255 patients

Abstract Background The purpose of this study was to investigate the long-term clinical outcome and its related factors regarding the severity of adhesion of CH ligament over long head of biceps (LHB) after shoulder arthroscopic capsular release for frozen shoulder with technical points in 255 patients. Methods We performed arthroscopic capsular release for frozen shoulder in 267 shoulders of 255 patients, 112 males and 143 females, with mean age of 56.39 years, mean disease duration periods of 0.934 years for conservative treatment, and mean follow-up periods of 5.6 years. The frozen shoulders were divided based on the severity of adhesion between CH ligament over LHB: those with slight degree of synovitis, no adhesion by obtuse rod, and slight thickness of the released capsule (type A), those with moderate degree of synovitis, moderate adhesion of the LHB by obtuse rod, and moderate thickness of the released capsule (type B), and those with severe degree of synovitis, severe adhesion of the LHB by obtuse rod, and severe thickness of the released capsule adhesion and a flatly shaped LHB (type C). We assessed the clinical factors related to the scoring of the shoulders by the criteria of the American Shoulder and Elbow Surgeons (ASES) and the relationship with severity of LHB adhesion. Results The ASES scores improved at 5 years postoperatively in all three groups significantly. The range of motion also significantly improved in all three groups significantly. The severity of the LHB adhesion over the CH ligament was confirmed to influence the ASES scores before and after the arthroscopic capsular release. There was a significant difference between type A and type B (p < 0.0001) or type C (p < 0.0001) before and after surgery. Logistic regression analysis showed disease duration, diabetes mellitus (DM), and ASES score were significantly associated to the severity type of LHB, especially DM has high odds ratio and was a risk factor for LHB adhesion. There is no adverse event including dislocation or axillary nerve injury and recurrence after arthroscopic capsular release at 5 years after surgery. Conclusions The long-term results of arthroscopic capsular release in frozen shoulder were confirmed in 255 patients. The severity of LHB adhesion over the CH ligament, a pathological condition related to DM as a risk factor, seems to play an important role in the functional outcome. Therefore, the sufficient release of LHB was essential technical point for arthroscopic capsular release in frozen shoulder

関節リウマチに対する関節鏡視下滑膜切除術におけるHo:YAGレーザー治療の効果

近年,関節リウマチ(RA)の治療においてレーザー治療が行われているが,その効果については明らかではない.RA患者11例13肩(stageI 1例,stageII 7例,stageIII 2例,stageIV 1例)に対して関節鏡視下滑膜切除術にHo:YAGレーザーを使用し,臨床的評価およびMRIによる評価を行った.RAの平均罹患期間は4.6年で経過観察期間は平均14ヵ月であった.さらに関節滑膜を初代培養しHo:YAGレーザーによる影響を細胞の形態学的およびサイトカイン産生をIL-1β,IL-6およびTNF-αについてELISAを用いて調べた.その結果,臨床的評価DAS28は術前平均5.4から術後平均3.7へ低下し,CRPは術前3.6から術後0.8に低下した. MRIでは13肩中10肩(77%)に滑脱増生の減少を認めた.滑膜細胞の形態は20WのHo:YAGレーザーにおいてアポトーシス様縮小を認めた.TNF-αは20Wまで増加を認めた.IL-1βとIL-6は15W以上において低下を認めた.以上より20W以上のHo:YAGレーザーにおいて滑膜細胞障害性が認められた.Ho:YAGレーザーは今後RA治療において滑膜細胞増殖を抑制させる手段として発展する可能性がある.To clarify the efficacy of holmium: yttrium-aluminium-garnet (Ho: YAG) laser therapy for arthroscopic synovectomy of rheumatoid arthritis (RA), we treated 13 shoulders of 11 RA patients of whom 1 was stage I, 7 stage II, 2 stage III, and 1 stage IV. The duration of RA is 4.6 years on average and the follow-up period is an average of 14 months. The Ho: YAG laser was set at 10 Watt (W) to treat the bone erosion areas so as to reach the deep zones of the pannus in order to resect the synovium. We compared C-reactive protein (CRP), Disease activity score (DAS) 28 and magnetic resonance image (MRI) findings before and after surgery. We cultured primary synovial cells to assay cytokine production of interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α. Morpho-logical examination was performed after treatment with the Ho: YAG laser at 0, 1, 5, 10, 15 and 20 W. We found villous synovium proliferation with vascularity in the rotator interval and supra spinatus tendon in the shoulder joints. In the subacromial bursa, yellow fat tissue and white fibrous soft tissue were detected in almost all shoulders. After synovectomy using the Ho: YAG laser, CRP decreased from an average of 3.6 to 0.8 and DAS28 also decreased from an average of 5.4 to 3.7 at 14 months after surgery. MRI showed decreased panni with synovium and did not precede joint destruction after 14 months in 10 shoulders out of 13 (77%). At 20 W of the Ho: YAG laser treatmentthe synovial cells shrank as in apoptosis and the number of cells also decreased. Laser treatment also resulted in the following significant changes: TNF-α production increased at 1, 10, 15 and 20 W (compared with 0 W) but not dose dependently; IL-1β and IL-6 increased up to 10W (compared with 0W) but decreased at 15 and 20 W (compared with 10 W). In morphological examination, after treatment with the Ho: YAG laser at 15 W, the synovial cells expanded and the number of cells decreased. Therefore, Ho: YAG laser therapy is effective for arthroscopic synovectomy, especially in treating pannus in bone erosion. We used 10 W for 5 seconds in each area where it could effectively decrease pannus formation

関節リウマチに対する関節鏡視下滑膜切除術におけるHo:YAGレーザー治療の効果

近年,関節リウマチ(RA)の治療においてレーザー治療が行われているが,その効果については明らかではない.RA患者11例13肩(stageI 1例,stageII 7例,stageIII 2例,stageIV 1例)に対して関節鏡視下滑膜切除術にHo:YAGレーザーを使用し,臨床的評価およびMRIによる評価を行った.RAの平均罹患期間は4.6年で経過観察期間は平均14ヵ月であった.さらに関節滑膜を初代培養しHo:YAGレーザーによる影響を細胞の形態学的およびサイトカイン産生をIL-1β,IL-6およびTNF-αについてELISAを用いて調べた.その結果,臨床的評価DAS28は術前平均5.4から術後平均3.7へ低下し,CRPは術前3.6から術後0.8に低下した. MRIでは13肩中10肩(77%)に滑脱増生の減少を認めた.滑膜細胞の形態は20WのHo:YAGレーザーにおいてアポトーシス様縮小を認めた.TNF-αは20Wまで増加を認めた.IL-1βとIL-6は15W以上において低下を認めた.以上より20W以上のHo:YAGレーザーにおいて滑膜細胞障害性が認められた.Ho:YAGレーザーは今後RA治療において滑膜細胞増殖を抑制させる手段として発展する可能性がある.To clarify the efficacy of holmium: yttrium-aluminium-garnet (Ho: YAG) laser therapy for arthroscopic synovectomy of rheumatoid arthritis (RA), we treated 13 shoulders of 11 RA patients of whom 1 was stage I, 7 stage II, 2 stage III, and 1 stage IV. The duration of RA is 4.6 years on average and the follow-up period is an average of 14 months. The Ho: YAG laser was set at 10 Watt (W) to treat the bone erosion areas so as to reach the deep zones of the pannus in order to resect the synovium. We compared C-reactive protein (CRP), Disease activity score (DAS) 28 and magnetic resonance image (MRI) findings before and after surgery. We cultured primary synovial cells to assay cytokine production of interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α. Morpho-logical examination was performed after treatment with the Ho: YAG laser at 0, 1, 5, 10, 15 and 20 W. We found villous synovium proliferation with vascularity in the rotator interval and supra spinatus tendon in the shoulder joints. In the subacromial bursa, yellow fat tissue and white fibrous soft tissue were detected in almost all shoulders. After synovectomy using the Ho: YAG laser, CRP decreased from an average of 3.6 to 0.8 and DAS28 also decreased from an average of 5.4 to 3.7 at 14 months after surgery. MRI showed decreased panni with synovium and did not precede joint destruction after 14 months in 10 shoulders out of 13 (77%). At 20 W of the Ho: YAG laser treatmentthe synovial cells shrank as in apoptosis and the number of cells also decreased. Laser treatment also resulted in the following significant changes: TNF-α production increased at 1, 10, 15 and 20 W (compared with 0 W) but not dose dependently; IL-1β and IL-6 increased up to 10W (compared with 0W) but decreased at 15 and 20 W (compared with 10 W). In morphological examination, after treatment with the Ho: YAG laser at 15 W, the synovial cells expanded and the number of cells decreased. Therefore, Ho: YAG laser therapy is effective for arthroscopic synovectomy, especially in treating pannus in bone erosion. We used 10 W for 5 seconds in each area where it could effectively decrease pannus formation

A CXC Chemokine Receptor, CXCR5/BLR1, Is a Novel and Specific Coreceptor for Human Immunodeficiency Virus Type 2

学位記番号:医博甲62

関節リウマチに対するインフリキシマブ治療による骨吸収マーカーの変化

近年,関節リウマチの治療に対して骨破壊抑制および改善効果のある生物学的製剤の使用により治療が行われている.しかし,すべての症例で骨破壊改善が認められるわけではなく,その詳細については現在不明である.当科でインフリキシマブにより治療し1年以上経過して骨代謝マーカーのデータが詳細にとれた12例について尿中NTxの改善を解析したので報告する.当科でインフリキシマブを使用した12例(男1例,女11例),平均年齢54.7(27～74)歳,平均罹患期間151ヵ月,平均MTX 5.5mg/week,平均Steroid 4.2mg/day,平均BMI31.4であった.投与前と投与後1年の尿中NTxクレアチニン換算値をWilcoxonの符号付き順位検定により比較し,投与後尿中NTxクレアチニン換算値の変化と年齢,罹患期間. steroid投与,リウマチ因子につきカイニ乗分析を行った.インフリキシマブ投与前の尿中NTxクレアチニン換算値は平均41.55±13.5から投与後平均40.77±16.34 n mol BCE/m mol CREであり有意差は認めなかった(p=0.814).投与後尿中NTxクレアチニン換算値の改善はリウマチ因子とステロイド投与に有意に関連した(p=0.038).すなわちリウマチ因子が低い,あるいはステロイド投与なしの症例では尿中NTxクレアチニン換算値は有意に低下を認めた.従って,リウマチ因子の低い,あるいはステロイド非投与の関節リウマチにおけるインフリキシマブ投与による治療は骨破壊改善を有意に導く可能性があると考えられた.In order to investigate which clinical factors are associated with bone improvement by treatment with infliximab in rheumatoid arthritis (RA), twelve cases using concise laboratory data were analyzed in terms of urinary NTx one year before and one year after treatment with infliximab. Urinary NTx changed from 41.55 ± 13.5 (20.9-62.8) (nM BCE/mM Cr) to 40.77 ± 16.34 (15-75.7) (nM BCE/mM Cr) and there was no significant difference between before and after treatment with infliximab (p=0.814). There was a significant correlation between improvement of urinary NTX and steroid or RAPA one year after treatment with infliximab (p=0.038 respectively). At less than 80 times RAPA, urinary NTx significantly decreased from 40.0 ± 15.0 (nM BCE/mM Cr) to 29.98 ± 8.65 (nM BCE/mM Cr) (p=0.043). Therefore, low RAPA and low dose of steroid was associated with improvement of urinary NTx after using infliximab. X-ray examination revealed that bone atrophy improved in 8 cases out of 12 (67%) and erosion improved in 1 case out of 12 (8.3%). MRI of the hand was assessed and synovium proliferation decreased in 1 case. Histological findings of the subchondral bone at the time of total elbow replacement during treatment with infliximab revealed newly formed fibrous woven bone, including osteoid material which filled the space of trabecular bone tissue. Therefore, the rheumatoid factor (RF) may be one of the points which indicates bone healing after using infliximab for RA