TRAIL and Taurolidine induce apoptosis and decrease proliferation in human fibrosarcoma

<p>Abstract</p> <p>Background</p> <p>Disseminated soft tissue sarcoma still represents a therapeutic dilemma because effective cytostatics are missing. Therefore we tested TRAIL and Tarolidine (TRD), two substances with apoptogenic properties on human fibrosarcoma (HT1080).</p> <p>Methods</p> <p>Viability, apoptosis and necrosis were visualized by TUNEL-Assay and quantitated by FACS analysis (Propidiumiodide/AnnexinV staining). Gene expression was analysed by RNA-Microarray and the results validated for selected genes by rtPCR. Protein level changes were documented by Western Blot analysis. NFKB activity was analysed by ELISA and proliferation assays (BrdU) were performed.</p> <p>Results and discussion</p> <p>The single substances TRAIL and TRD induced apoptotic cell death and decreased proliferation in HT1080 cells significantly. Gene expression of several genes related to apoptotic pathways (TRAIL: <it>ARHGDIA</it>, <it>NFKBIA</it>, <it>TNFAIP3</it>; TRD: <it>HSPA1A/B</it>, <it>NFKBIA</it>, <it>GADD45A</it>, <it>SGK</it>, <it>JUN</it>, <it>MAP3K14</it>) was changed. The combination of TRD and TRAIL significantly increased apoptotic cell death compared to the single substances and lead to expression changes in a variety of genes (<it>HSPA1A/B</it>, <it>NFKBIA</it>, <it>PPP1R15A</it>, <it>GADD45A</it>, <it>AXL</it>, <it>SGK</it>, <it>DUSP1</it>, <it>JUN</it>, <it>IRF1</it>, <it>MYC</it>, <it>BAG5</it>, <it>BIRC3</it>). NFKB activity assay revealed an antipodal regulation of the several subunits of NFKB by TRD and TRD+TRAIL compared to TRAIL alone.</p> <p>Conclusion</p> <p>TRD and TRAIL are effective to induce apoptosis and decrease proliferation in human fibrosarcoma. A variety of genes seems to be involved, pointing to the NFKB pathway as key regulator in TRD/TRAIL-mediated apoptosis.</p

Apoptoseinduktion und Proliferationshemmung durch Taurolidin und TRAIL in humanen Fibrosarkomzellen (HT1080)

Für Weichgewebssarkome fehlt nach wie vor eine effektive Behandlung über die Resektion hinaus. Wir untersuchten die Effekte von TRAIL und Taurolidin (TRD) auf humane Fibrosarkomzellen (HT1080). Apoptoseinduktion und Proliferationshemmung nach Inkubation mit den Substanzen wurden im TUNEL- bzw. Proliferationsassay dargestellt und mittels Durchflusszytometrie quantifiziert. Veränderungen der Gen- bzw. Proteinexpression analysierten wir mittels RNA-Microarray, für einzelne Gene auch in der Real-time-PCR und im Western-Blot. Die Aktivität des Transkriptionsfaktors NF$_{K}$B wurde mittels ELISA dargestellt. TRAIL und TRD induzierten signifikant Apoptose, reduzierten die Proliferation der Zellen und veränderten die Expression verschiedener apoptoseassoziierter Gene mit Hinweisen auf den NF$_{K}$B-Signalweg als Schlüsselfaktor - die Kombination dabei effektiver als die Einzelsubstanzen. Das NF$_{K}$B-Assay zeigte eine entgegengesetzte Regulation der Untereinheiten des Transkriptionsfaktors

Cost of vaccine delivery strategies in low- and middle-income countries during the COVID-19 pandemic

BACKGROUND: The COVID-19 pandemic has disrupted immunization services critical to the prevention of vaccine-preventable diseases in many low- and middle- income countries around the world. These services will need to be modified in order to minimize COVID-19 transmission and ensure the safety of health workers and the community. Additional budget will be required to implement these modifications that ensure safe delivery. METHODS: Using a simple modeling analysis, we estimated the additional resource requirements associated with modifications to supplementary immunization activities (campaigns) and routine immunization services via fixed sites and outreach in 2020 US dollars. We considered the following four categories of costs: (1) personal protective equipment (PPE) &amp; infection prevention and control (IPC) measures for immunization sessions; (2) physical distancing and screening during immunization sessions; (3) delivery strategy changes, such as changes in session sizes and frequency; and (4) other operational cost increases, including additional social mobilization, training, and hazard pay to compensate health workers. RESULTS: We found that implementing a range of measures to protect health workers and communities from COVID-19 transmission could result in a per-facility start-up cost of $466–799 for routine fixed-site delivery and$12–220 for routine outreach delivery, and $12–108 per immunization campaign site. A recurrent monthly cost of$137–1,024 for fixed-site delivery and $152–848 for outreach delivery per facility could be incurred, and a$0.32–0.85 increase in the cost per dose during campaigns. CONCLUSIONS: By illustrating potential cost implications of providing immunization services through a range of strategies in a safe manner, these estimates can provide a benchmark for program managers and policy makers on the additional budget required. These findings can help country practitioners and global development partners planning the continuation of immunization services in the context of COVID-19