Autoreactive B cells in rheumatoid arthritis

Rheumatoid arthritis (RA) is an autoimmune disease affecting joints which is hallmarked by the presence of autoantibodies against citrrulianted protein (ACPA). This thesis describes the phenotypic and functional characteristics of ACPA-expressing autoreactive B cells which suggest potential pathologic roles of these B cells in RA pathogenesis. The thesis also describes strategies to specifically deplete ACPA-expressing B cells to improve current RA therapeutic options.The Institute for Chemical Immunology, an NWO Gravitational programLUMC / Geneeskund

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer‐reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state‐of‐the‐art handbook for basic and clinical researchers.DFG, 389687267, Kompartimentalisierung, Aufrechterhaltung und Reaktivierung humaner Gedächtnis-T-Lymphozyten aus Knochenmark und peripherem BlutDFG, 80750187, SFB 841: Leberentzündungen: Infektion, Immunregulation und KonsequenzenEC/H2020/800924/EU/International Cancer Research Fellowships - 2/iCARE-2DFG, 252623821, Die Rolle von follikulären T-Helferzellen in T-Helferzell-Differenzierung, Funktion und PlastizitätDFG, 390873048, EXC 2151: ImmunoSensation2 - the immune sensory syste

Implementasi Material Handling dalam Mencari Jarak dan Ongkos Material serta Usulan Tata Letak Produksi di PT. Wijaya Karya Beton

Tata letak dalam fasilitas produksi harus direncanakan dengan baik dan jarak dibuat seefisien mungkin antara unit kerja agar tidak terjadinya peningkatan biaya. Jarak untuk aliran lintasan di PT Wijaya Karya Beton saat ini belum diketahui efisien atau tidak dengan jarak tempuh keseluruhan departemen sepanjang 300 meter, dan total biaya yaitu Rp 60.000/jam. Material Handling ialah salah satu jenis transportasi (pengangkutan) yang dilakukan dalam perusahaan industri, yang artinya memindahkan bahan baku, barang setengah jadi atau barang jadi dari tempat asal ke tempat tujuan yang telah ditetapkan. Penelitian ini ditujukan untuk mengetahui perancangan fasilitas yang cocok pada pabrik dengan dibuat layout produksi berdasarkan keterkaitan tiap-tiap departemen dengan alat material handling yaitu crane. Dalam mengetahui layout seperti apa yang baik ialah mencari ongkos material handling terlebih dahulu. Dalam hasil perhitungan diperoleh ongkos material handling yang dikeluarkan yaitu Rp. 200/m. Selanjutnya mencari jarak satu departemen ke departemen lain, ongkos tiap departemen, membuat Flow to Chart (FTC), membuat Tabel Skala Prioritas (TSP) dan yang terakhir ialah membuat Activity Relationship Diagram (ARD) dengan berdasarkan dari hasil FTC tersebut. Hasil penelitian ini menunjukan bahwa urutan aliran antar departemen pada lantai produksi yaitu A-B-C-D-E dengan layout yang dibuat berbentuk U flow. Berdasarkan hasil penelitian, layout usulan dinilai sudah efektif dan efisien karena disusun berdasarkan jarak antar departemen dan ongkos material handling pada lantai produksi

Collaboration of high activity soil and geological structure factors in Pagelaran soil creep occurrence, Indonesia

Soil creep is a landslide that moves slowly but it does not mean harmless. Soil creep that succeeded in forcing the citizens to migrate occurred in Pagelaran by wiping out 13 houses. This paper aims to know the contribution of geological structures and physical properties of soil toward the creeping occurrence. The methods used in this research consist of geological structure mapping; soil sampling; laboratory works; and analysis of geological structures and soil activity. Geological structures analysis indicates that the main force worked predominantly in NE-SW direction, parallel with creeping. It expresses that the creep was influenced by the orientation of them. Additionally, their distribution can be used as access for rainwater to infiltrate and add to the burden of the slopes. While laboratory results show that, the soil is classified into MH and CH. Calculation of activity numbers of soil indicates that it has a high – very high activity with value between 0.705 – 1.4931 (Skempton, 1953). It means that the soil will swell easily if contaminated by water

Collaboration of high activity soil and geological structure factors in Pagelaran soil creep occurrence, Indonesia

Soil creep is a landslide that moves slowly but it does not mean harmless. Soil creep that succeeded in forcing the citizens to migrate occurred in Pagelaran by wiping out 13 houses. This paper aims to know the contribution of geological structures and physical properties of soil toward the creeping occurrence. The methods used in this research consist of geological structure mapping; soil sampling; laboratory works; and analysis of geological structures and soil activity. Geological structures analysis indicates that the main force worked predominantly in NE-SW direction, parallel with creeping. It expresses that the creep was influenced by the orientation of them. Additionally, their distribution can be used as access for rainwater to infiltrate and add to the burden of the slopes. While laboratory results show that, the soil is classified into MH and CH. Calculation of activity numbers of soil indicates that it has a high – very high activity with value between 0.705 – 1.4931 (Skempton, 1953). It means that the soil will swell easily if contaminated by water

Rap1 Can Bypass the FAK-Src-Paxillin Cascade to Induce Cell Spreading and Focal Adhesion Formation

We developed new image analysis tools to analyse quantitatively the extracellular-matrix-dependent cell spreading process imaged by live-cell epifluorescence microscopy. Using these tools, we investigated cell spreading induced by activation of the small GTPase, Rap1. After replating and initial adhesion, unstimulated cells exhibited extensive protrusion and retraction as their spread area increased, and displayed an angular shape that was remodelled over time. In contrast, activation of endogenous Rap1, via 007-mediated stimulation of Epac1, induced protrusion along the entire cell periphery, resulting in a rounder spread surface, an accelerated spreading rate and an increased spread area compared to control cells. Whereas basal, anisotropic, spreading was completely dependent on Src activity, Rap1-induced spreading was refractory to Src inhibition. Under Src inhibited conditions, the characteristic Src-induced tyrosine phosphorylations of FAK and paxillin did not occur, but Rap1 could induce the formation of actomyosin-connected adhesions, which contained vinculin at levels comparable to that found in unperturbed focal adhesions. From these results, we conclude that Rap1 can induce cell adhesion and stimulate an accelerated rate of cell spreading through mechanisms that bypass the canonical FAK-Src-Paxillin signalling cascade

Live-cell imaging of 007-induced adhesion and spreading.

<p>A549-Epac1 cells expressing GFP-Lifeact were trypsinised and allowed to roll for 1 hour before plating out on fibronectin with or without 100 µM 007. Images of cells were captured through the 63×objective every 5 minutes from 30 minutes after plating, with representative images shown in (A). The scale bars represent 10 µm. Altogether, 54 cells without 007 and 41 cells with 007 from 10 separate spreading experiments were quantified. The spread area at each frame was determined in pixels and standardised to the average area of all the first frames of the basal (−007) spreading cells (B). The spreading rate over the first 30 minutes of imaging was calculated using the gradient (m) of the red lines show on each graph and is presented in spread area per hour. Dashed vertical lines indicate the time at which cells approached their maximal area. Area dynamics (C) were determined by measuring the gain or loss of spread area between consecutive frames, and presented as a proportion of the spread area of the second frame. The dashed vertical lines show the time at which the area of protrusion equalled the area of retraction. Cell morphology was analysed by the circle ratio (D), which was calculated by dividing the area of the cell at each time point by the area of a circle which encompassed the outer-most tips of the detected cell edges. Changes in the shape of the spread area of cells were calculated by determining the normalised variance (σ_n value) of the distances from the cell centre to each peripheral point of the cell. Then, the change in the variance (Δσ_n value) between consecutive time points was calculated (E). For all graphs shown, each plotted point is the mean of all cells measured ± the standard error of the mean.</p

Effects of the Src family kinase inhibitor, PP2, on basal and 007-induced cell spreading.

<p>Cells were trypsinised, rolled for 1 hour without or with 20 µM PP2 pre-treatment and plated on fibronectin with or without 007. Adhesion of A549-Epac1 cells after 30 minutes was determined by alkaline phosphatase activity (A). PP2-treated A549-Epac1-GFP-Lifeact cells were plated on fibronectin and imaged over time using the 63×objective. Shown are representative images (B) and quantification of the mean spread area ± the standard error of the means (C) of 16 cells plated without 007 and 15 cells with 100 µM 007 from three separate spreading experiments. Spread area was standardised to the average area of −007 cells shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0050072#pone-0050072-g001" target="_blank">Figure 1B</a> at the first frame of imaging (from assays performed independently, but under comparable experimental conditions). The dashed grey curve shows the spreading kinetics of 007-treated cells from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0050072#pone-0050072-g001" target="_blank">Figure 1B</a>. The vertical line indicates the time that cells reached their maximum spread area. The spreading rate was calculated using the gradient (m), in spread area per hour, of the red line shown on the graph. In (D) and (E), A549-Epac1-GFP-Lifeact-expressing cells captured through the 20×objective spread in the presence or absence of 100 µM 007 for 2.5 hours before 20 µM PP2 was added. For quantification (E), the spread area was standardised to the average area of −007 cells at the first time point of imaging. In (F), human umbilical vein endothelial cells were plated on fibronectin for 1 hour in the presence and absence of 100 µM 007 and 20 µM PP2, before being fixed and stained. At least 30 cells per condition were quantified and standardised to the mean area of cells spreading without 007 or PP2. In (A) and (F), graphs show the mean of 5 and 3 experiments, respectively ± the standard error of the means. <i>P</i> values were calculated by paired student’s t-tests.</p