Urokinase-type plasminogen activator and arthritis progression: contrasting roles in systemic and monoarticular arthritis models

INTRODUCTION: Urokinase-type plasminogen activator (u-PA) has been implicated in tissue destruction/remodeling. The absence of u-PA results in resistance of mice to systemic immune complex-driven arthritis models; monoarticular arthritis models involving an intra-articular (i.a.) antigen injection, on the other hand, develop more severe arthritis in its absence. The aims of the current study are to investigate further these contrasting roles that u-PA can play in the pathogenesis of inflammatory arthritis and to determine whether u-PA is required for the cartilage and bone destruction associated with disease progression. METHODS: To determine how the different pathogenic mechanisms leading to arthritis development in the different models may explain the contrasting requirement for u-PA, the systemic, polyarticular, immune complex-driven K/BxN arthritis model was modified to include an i.a. injection of saline as a local trauma in u-PA-/- mice. This modified model and the antigen-induced arthritis (AIA) model were also used in u-PA-/- mice to determine the requirement for u-PA in joint destruction. Disease severity was determined by clinical and histologic scoring. Fibrin(ogen) staining and the matrix metalloproteinase (MMP)-generated neoepitope DIPEN staining were performed by immunohistochemistry. Gene expression of inflammatory and destructive mediators was measured in joint tissue by quantitative PCR. RESULTS: In our modified arthritis model, u-PA-/- mice went from being resistant to arthritis development following K/BxN serum transfer to being susceptible following the addition of an i.a. injection of saline. u-PA-/- mice also developed more sustained AIA compared with C57BL/6 mice, including reduced proteoglycan levels and increased bone erosions, fibrin(ogen) deposition and DIPEN expression. Synovial gene expression of the proinflammatory mediators (TNF and IL-1β), aggrecanases (ADAMTS-4 and -5) and MMPs (MMP3 and MMP13) were all sustained over time following AIA induction in u-PA-/- mice compared with C57BL/6 mice. CONCLUSIONS: We propose that u-PA has a protective role in arthritis models with 'wound healing-like' processes following local trauma, possibly through u-PA/plasmin-mediated fibrinolysis, but a deleterious role in systemic models that are critically dependent on immune complex formation and complement activation. Given that cartilage proteoglycan loss and bone erosions were present and sustained in u-PA-/- mice with monoarticular arthritis, it is unlikely that u-PA/plasmin-mediated proteolysis is contributing directly to this tissue destruction/remodeling

高効率線形送信法におけるPWM包絡線発生法の研究

電気通信大学200

Urokinase-type plasminogen activator and arthritis progression: role in systemic disease with immune complex involvement

INTRODUCTION: Urokinase-type plasminogen activator (u-PA) has been implicated in fibrinolysis, cell migration, latent cytokine activation, cell activation, T-cell activation, and tissue remodeling, all of which are involved in the development of rheumatoid arthritis. Previously, u-PA has been reported to play a protective role in monoarticular arthritis models involving mBSA as the antigen, but a deleterious role in the systemic polyarticular collagen-induced arthritis (CIA) model. The aim of the current study is to determine how u-PA might be acting in systemic arthritis models. METHODS: The CIA model and bone marrow chimeras were used to determine the cellular source of u-PA required for the arthritis development. Gene expression of inflammatory and destructive mediators was measured in joint tissue by quantitiative PCR and protein levels by ELISA. The requirement for u-PA in the type II collagen mAb-induced arthritis (CAIA) and K/BxN serum transfer arthritis models was determined using u-PA(-/-) mice. Neutrophilia was induced in the peritoneal cavity using either ovalbumin/anti-ovalbumin or the complement component C5a. RESULTS: u-PA from a bone marrow-derived cell was required for the full development of CIA. The disease in u-PA(-/-) mice reconstituted with bone marrow from C57BL/6 mice was indistinguishable from that in C57BL/6 mice, in terms of clinical score, histologic features, and protein and gene expression of key mediators. u-PA(-/-) mice were resistant to both CAIA and K/BxN serum transfer arthritis development. u-PA(-/-) mice developed a reduced neutrophilia and chemokine production in the peritoneal cavity following ovalbumin/anti-ovalbumin injection; in contrast, the peritoneal neutrophilia in response to C5a was u-PA independent. CONCLUSIONS: u-PA is required for the full development of systemic arthritis models involving immune complex formation and deposition. The cellular source of u-PA required for CIA is bone marrow derived and likely to be of myeloid origin. For immune complex-mediated peritonitis, and perhaps some other inflammatory responses, it is suggested that the u-PA involvement may be upstream of C5a signaling

Determining the actual cost of wound care in Australia

Aim: To determine the number and type of wounds and their treatment costs (consumables and labour) in Australian hospitals, residential aged care facilities (RACFs), general practices (GPs) and community, and to provide evidence to inform reimbursement of wound treatment costs for all Australians. Method: Data from 21,189 clients with 49,234 wounds treated by a community care provider in Western Australia, Queensland and South Australia during the financial year 2020/2021 were used to determine the mean and median costs (consumables and labour) to treat wounds. Surveys involving skin inspections and medical record audits were conducted amongst consenting adults over 18 years old in a sample of Australian hospitals, RACFs and GPs. A sample of community clients’ data for wounds treated on one day in June 2021 comprised the fourth cohort used in this analysis. The costs to treat all wounds surveyed between 14 December 2020 and 17 October 2021 in the four cohorts were modelled against the community care provider’s data for 2020/2021 (49,234 wounds). Results: There were 2,505 individuals with 3,096 wounds. The estimated cost to treat all wounds was A$1,621,768 using the mean costs of the community care provider as a basis, and A$692,144 using the median costs of the community care provider as a basis. Costs for all wound types were determined. Conclusion: The cost of treating wounds in Australia was determined and is anticipated to inform a review of equitable reimbursement of wound treatment costs for Australians with wounds.</p

無線LANアクセスポイントの検索要求を用いた屋内混雑度推定手法

ある場所がどの時間にどれだけ混雑しているか、という混雑情報はその場所の事業者にとっても利用者にとっても有益である。例えば、鉄道の混雑度(乗車率)を広く収集し分析を行えば空いている電車を優先的に検索する路線検索システムを作ることができるし、店舗で混雑度(来客者数)を収集すれば、来客者のうち何人が実際に商品を購入したかという分析を行うことができる。もっとも広く普及している混雑度の測定方法は目視測定であるが、人件費もかかり長期間データを収集することが難しいという欠点がある。混雑度測定の自動化方式としては、カメラを用いて導線解析する方式や二酸化炭素センサを用いる方式などがあるが、適用できるエリアが限定的であり、広く使われるには至っていない。そこで本研究では近年普及してきた無線LAN搭載モバイル端末に着目する。無線LAN端末は、周辺のアクセスポイントを検索するために不定期にプローブ要求をブロードキャストしている。この信号は端末が自然に送出するものであり、端末の特別な操作や事前の準備は必要ない。本研究では、この信号を利用し無線LAN信号のキャプチャ機器を設置するだけで混雑度の推定が行えるシステムを提案する。実際に鉄道列車内と大学の教室内で実験を行い、鉄道列車内では混雑度に対して相関係数0.74の特徴量を、教室内では在室人数に対して相関係数0.86の特徴量を得られることを示し た。電気通信大学201

Physicochemical and Immunological Assessment of Engineered Pure Protein Particles with Different Redox States

The development of subunit antigen delivery formulations has become an important research endeavor, especially in cases where a whole cell vaccine approach has significant biosafety issues. Particle-based systems have shown particular efficacy due to their inherent immunogenicity. In some cases, fabrication techniques can lead to changes in the redox states of encapsulated protein antigens. By employing a uniform, well-characterized, single-protein system, it is possible to elucidate how the molecular details of particle-based protein antigens affect their induced immune responses. Using mesoporous silica-templated, amide bond-stabilized ovalbumin particles, three types of particles were fabricated from native, reduced, and oxidized ovalbumin, resulting in particles with different physicochemical properties and immunogenicity. Phagocytosis, transcription factor activation, and cytokine secretion by a mouse macrophage cell line did not reveal significant differences between the three types of particles. Oxidation of the ovalbumin, however, was shown to inhibit the intracellular degradation of the particles compared with native and reduced ovalbumin particles. Slow intracellular degradation of the oxidized particles was correlated with inefficient antigen presentation and insignificant levels of T cell priming and antibody production <i>in vivo</i>. In contrast, particles fabricated from native and reduced ovalbumin were rapidly degraded after internalization by macrophages <i>in vitro</i> and resulted in significant T cell and B cell immune responses <i>in vivo</i>. Taken together, the current study demonstrates how the redox state of a protein antigen significantly impacts the immunogenicity of the particulate vaccine formulations

Cytokine mRNA and nitrite levels obtained from naive, M1 and M2 macrophages.

<p>Cytokine primed macrophages were incubated with <i>P</i>. <i>gingivalis</i> W50 (Bacteria to cell ratio 100:1) for 90 minutes at 37°C and supernatant was collected 24 hours post infection for nitric oxide (nitrite metabolite) and cells were lysed for mRNA extraction. <b>(A)</b> Cytokine concentration was determined using RT-PCR. <b>(B) N</b>itric oxide (NO) secretion by naive, M1 and M2 macrophages was measured indirectly by quantifying the level of nitrite (metabolite) in cell supernatants. Cytochalasin B was added as negative control as it stabilises microtubules and inhibit actin polymerisation, a process required for phagocytosis. Data are representative of independent experiments and triplicate biological samples are expressed as the mean ± standard deviation (n = 3) and were analysed using student's t-test. * indicates data that are significantly different (<i>p</i> < 0.05) from the data for non-stimulated group (media and macrophage alone group).</p

3D-SIM images of <i>P</i>. <i>gingivalis</i> W50 phagocytosed by naive, M1 and M2 macrophages.

<p><b>(A,B,C)</b> Maximum intensity projection and <b>(D,E,F)</b> the corresponding slice view 3D-SIM images of <i>P</i>. <i>gingivalis</i> W50 phagocytosed by naive, M1 and M2 macrophages (60:1 BMR, 1 hour exposure time), respectively, on the Deltavision OMX Structured Illumination Microscope V4 Blaze (Applied Precision, WA, USA). <i>P</i>. <i>gingivalis</i> W50 were labelled with AF488 (green) while the nuclei and actin filaments of the macrophages were stained with DAPI (blue) and Phalloidin-TRITC (red) respectively. The slice images were obtained from a section with defined size (between 10–50 z-stack) to confirm phagocytosis of <i>P</i>. <i>gingivalis</i> W50 by naive, M1 and M2 macrophages.</p

The effect of concentration of <i>P</i>. <i>gingivalis</i> on naive, M1 and M2 macrophage apoptosis.

<p>The percentage of apoptotic naive, M1 and M2 macrophages after incubation with <i>P</i>. <i>gingivalis</i> (bacteria to macrophage ratio– 10: 1, 100:1 and 1000:1) for 90 minutes, 37°C. The negative controls were naive, M1 and M2 macrophages alone. Apoptosis was detected using YO-PRO®-1 dyes and flow cytometric analysis. Data are representative of independent experiments and triplicate biological samples are expressed as the mean ± standard deviation (n = 3) and were analysed using student's t-test. * indicates data that are significantly different (<i>p</i> < 0.05) from the data for non-stimulated group (macrophage alone group).</p