A Review of Obinutuzumab (GA101), a Novel Type II Anti-CD20 Monoclonal Antibody, for the Treatment of Patients with B-Cell Malignancies

<p><strong>Article full text</strong></p> <p><br> The full text of this article can be found <a href="https://link.springer.com/article/10.1007/s12325-016-0451-1"><b>here</b>.</a><br> <br> <strong>Provide enhanced digital features for this article</strong><br> If you are an author of this publication and would like to provide additional enhanced digital features for your article then please contact <u>[email protected]</u>.<br> <br> The journal offers a range of additional features designed to increase visibility and readership. All features will be thoroughly peer reviewed to ensure the content is of the highest scientific standard and all features are marked as ‘peer reviewed’ to ensure readers are aware that the content has been reviewed to the same level as the articles they are being presented alongside. Moreover, all sponsorship and disclosure information is included to provide complete transparency and adherence to good publication practices. This ensures that however the content is reached the reader has a full understanding of its origin. No fees are charged for hosting additional open access content.<br> <br> Other enhanced features include, but are not limited to:<br> • Slide decks<br> • Videos and animations<br> • Audio abstracts<br> • Audio slides<u></u></p

Expression patterns of DMEM- and MC-XF-cultured JPCs (growing within coated flasks) by flow cytometric analyses.

<p>Representative histograms and the average percentages (±STD) of positive cells for CD29, CD45, CD73, CD90 and CD105 expression by unseparated JPCs are illustrated.</p

Quantitative analysis of gene expression levels in DMEM- and MC-XF-cultured JPCs at day 5 and 10 of osteogenesis (of passage 6, n = 4).

<p>Induction indices (x-fold) and significance values of alkaline phosphatase, Runx-2, type I collagen (alpha1-chain) and osteoprotegerin in osteogenic induced in comparison to untreated cells under both culture conditions are illustrated.</p

Quantification of MSCA-1⁺ cells under DMEM- and MC-XF culturing conditions by FACS analysis.

<p>Unseparated JPCs were plated in culture dishes in DMEM medium. For the first test runs, cells were maintained in DMEM medium whereas the second series underwent stepwise FCS reduction and convertion to MC-XF medium. At the same time points (day 2, 5 and 9 after conversion from DMEM to MC-XF culture conditions), cells were detached from the dishes and the percentages of MSCA-1⁺ cells were determined by FACS analysis. Significant higher amounts of MSCA-1⁺ cells were detected under MC-XF culturing conditions at day 2 and 5 (p<0.01 and p<0.05).</p

MSCA-1 expression (% positive cells ± STD).

<p>Overview of average percentages (± STD) of MSCA-1⁺ cells under DMEM and MC-XF culture conditions (n  =  5) as detected by FACS analysis. Flow cytometric analyses were carried out at day 2, 5 and 9 immediately after conversion from DMEM to MC-XF culture conditions.</p

PDT (days ± STD) during in vitro passaging.

<p>Overview of calculated average population doubling times (PDT, in days, ± STD) in JPCs cultured under DMEM and MC-XF conditions (n = 3 or n = 4, as indicated in the table) at different passages (passage 4–7).</p

Detection of mineral deposition by non-mineralizing JPCs (of passage 6).

<p>Representative fluorescent staining of hydroxyapatite formation by OsteoImage in DMEM- and MC-XF-cultured monolayers (growing within coated flasks) at day 20 of osteogenesis. Non-mineralizing JPCs showed mineralization capacity only under MC-XF but not DMEM media conditions. 10× magnification. Co =  untreated cell, ob =  osteogenic induced cells.</p

Life-monitoring measurements of cell proliferation by unseparated and MSCA-1-separated JPCs using the x-CELLigence system (representative curve progression of cells derived from one donor).

<p>Cells (of passage 5) were seeded into E-plates for the life-measurements of cell impedance following the same experimental procedures as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0081674#pone-0081674-g001" target="_blank">fig. 1</a> (gradual FCS reduction and addition of the MC-XF culture medium). Note the preferential proliferation activation of the MSCA-1⁺ subpopulation (pink line) in contrast to the rather restrictive capacity of the xenogeneic-free culture medium on MSCA-1⁻ JPCs (violet line). The lower panel of the figure illustrates representative histograms of MSCA-1 expression by flow cytometry before (unsep.) or after the magnetic separation of the MSCA-1+/− fractions.</p

Life-monitoring measurements of cell proliferation by unseparated JPCs using the x-CELLigence system (ACEA Biosciences).

<p>JPCs of passage 4 derived from two different donors were seeded into special E-plates in DMEM/F12/10%FCS culture medium. Two days later (44 hours - each tick of the scale corresponds to 11 hours), a gradual FCS reduction was performed in one-half of the test runs (green and dark green), whereas the other half of the wells was further cultivated in DMEM/F12/10%FCS (red and coral). Nine days (297 hours) after the initiation of FCS reduction, MC-XF culture medium was added to the cells. The proliferation curve progression of DMEM-cultured JPCs (from two representative patients) is highlighted in red and coral and that of MC-XF-cultivated cells is highlighted in dark green and green. The right panel of the figure shows the cell morphology of DMEM- and MC-XF-cultured JPCs. Note the reduced cell size under MC-XF culture conditions (on uncoated dishes) leading to the significant decrease of cell impedance immediately after the addition of the MC-XF culture medium.</p

Xanthurenic Acid Binds to Neuronal G-Protein-Coupled Receptors That Secondarily Activate Cationic Channels in the Cell Line NCB-20

<div><p>Xanthurenic acid (XA) is a metabolite of the tryptophan oxidation pathway through kynurenine and 3-hydroxykynurenine. XA was until now considered as a detoxification compound and dead-end product reducing accumulation of reactive radical species. Apart from a specific role for XA in the signaling cascade resulting in gamete maturation in mosquitoes, nothing was known about its functions in other species including mammals. Based upon XA distribution, transport, accumulation and release in the rat brain, we have recently suggested that XA may potentially be involved in neurotransmission/neuromodulation, assuming that neurons presumably express specific XA receptors. Recently, it has been shown that XA could act as a positive allosteric ligand for class II metabotropic glutamate receptors. This finding reinforces the proposed signaling role of XA in brain. Our present results provide several lines of evidence in favor of the existence of specific receptors for XA in the brain. First, binding experiments combined with autoradiography and time-course analysis led to the characterization of XA binding sites in the rat brain. Second, specific kinetic and pharmacological properties exhibited by these binding sites are in favor of G-protein-coupled receptors (GPCR). Finally, in patch-clamp and calcium imaging experiments using NCB-20 cells that do not express glutamate-induced calcium signals, XA elicited specific responses involving activation of cationic channels and increases in intracellular Ca²⁺ concentration. Altogether, these results suggest that XA, acting through a GPCR-induced cationic channel modulatory mechanism, may exert excitatory functions in various brain neuronal pathways.</p> </div