Alternating Electric Fields (Tumor-Treating Fields Therapy) Can Improve Chemotherapy Treatment Efficacy in Non-Small Cell Lung Cancer Both In Vitro and In Vivo

Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer-related deaths worldwide. Common treatment modalities for NSCLC include surgery, radiotherapy, chemotherapy, and, in recent years, the clinical management paradigm has evolved with the advent of targeted therapies. Despite such advances, the impact of systemic therapies for advanced disease remains modest, and as such, the prognosis for patients with NSCLC remains poor. Standard modalities are not without their respective toxicities and there is a clear need to improve both efficacy and safety for current management approaches. Tumor-treating fields (TTFields) are low-intensity, intermediate-frequency alternating electric fields that disrupt proper spindle microtubule arrangement, thereby leading to mitotic arrest and ultimately to cell death. We evaluated the effects of combining TTFields with standard chemotherapeutic agents on several NSCLC cell lines, both in vitro and in vivo. Frequency titration curves demonstrated that the inhibitory effects of TTFields were maximal at 150 kHz for all NSCLC cell lines tested, and that the addition of TTFields to chemotherapy resulted in enhanced treatment efficacy across all cell lines. We investigated the response of Lewis lung carcinoma and KLN205 squamous cell carcinoma in mice treated with TTFields in combination with pemetrexed, cisplatin, or paclitaxel and compared these to the efficacy observed in mice exposed only to the single agents. Combining TTFields with these therapeutic agents enhanced treatment efficacy in comparison with the respective single agents and control groups in all animal models. Together, these findings suggest that combining TTFields therapy with chemotherapy may provide an additive efficacy benefit in the management of NSCLC

Effect of natalizumab on disease progression in secondary progressive multiple sclerosis (ASCEND). a phase 3, randomised, double-blind, placebo-controlled trial with an open-label extension

Background: Although several disease-modifying treatments are available for relapsing multiple sclerosis, treatment effects have been more modest in progressive multiple sclerosis and have been observed particularly in actively relapsing subgroups or those with lesion activity on imaging. We sought to assess whether natalizumab slows disease progression in secondary progressive multiple sclerosis, independent of relapses. Methods: ASCEND was a phase 3, randomised, double-blind, placebo-controlled trial (part 1) with an optional 2 year open-label extension (part 2). Enrolled patients aged 18–58 years were natalizumab-naive and had secondary progressive multiple sclerosis for 2 years or more, disability progression unrelated to relapses in the previous year, and Expanded Disability Status Scale (EDSS) scores of 3·0–6·5. In part 1, patients from 163 sites in 17 countries were randomly assigned (1:1) to receive 300 mg intravenous natalizumab or placebo every 4 weeks for 2 years. Patients were stratified by site and by EDSS score (3·0–5·5 vs 6·0–6·5). Patients completing part 1 could enrol in part 2, in which all patients received natalizumab every 4 weeks until the end of the study. Throughout both parts, patients and staff were masked to the treatment received in part 1. The primary outcome in part 1 was the proportion of patients with sustained disability progression, assessed by one or more of three measures: the EDSS, Timed 25-Foot Walk (T25FW), and 9-Hole Peg Test (9HPT). The primary outcome in part 2 was the incidence of adverse events and serious adverse events. Efficacy and safety analyses were done in the intention-to-treat population. This trial is registered with ClinicalTrials.gov, number NCT01416181. Findings: Between Sept 13, 2011, and July 16, 2015, 889 patients were randomly assigned (n=440 to the natalizumab group, n=449 to the placebo group). In part 1, 195 (44%) of 439 natalizumab-treated patients and 214 (48%) of 448 placebo-treated patients had confirmed disability progression (odds ratio [OR] 0·86; 95% CI 0·66–1·13; p=0·287). No treatment effect was observed on the EDSS (OR 1·06, 95% CI 0·74–1·53; nominal p=0·753) or the T25FW (0·98, 0·74–1·30; nominal p=0·914) components of the primary outcome. However, natalizumab treatment reduced 9HPT progression (OR 0·56, 95% CI 0·40–0·80; nominal p=0·001). In part 1, 100 (22%) placebo-treated and 90 (20%) natalizumab-treated patients had serious adverse events. In part 2, 291 natalizumab-continuing patients and 274 natalizumab-naive patients received natalizumab (median follow-up 160 weeks [range 108–221]). Serious adverse events occurred in 39 (13%) patients continuing natalizumab and in 24 (9%) patients initiating natalizumab. Two deaths occurred in part 1, neither of which was considered related to study treatment. No progressive multifocal leukoencephalopathy occurred. Interpretation: Natalizumab treatment for secondary progressive multiple sclerosis did not reduce progression on the primary multicomponent disability endpoint in part 1, but it did reduce progression on its upper-limb component. Longer-term trials are needed to assess whether treatment of secondary progressive multiple sclerosis might produce benefits on additional disability components. Funding: Biogen

Classification of Events Violating the Safety of Physical Layers in Fiber-Optic Network Infrastructures

Fiber-optic network infrastructures are crucial for the transmission of data over long and short distances. Fiber optics are also preferred for the infrastructure of in-building data communications. In this study, we use polarization analysis to ensure the security of the optical fiber/cables of the physical layer. This method exploits the changes induced by mechanical vibrations to polarization states, which can be easily detected using a polarization beam splitter and a balancing photodetector. We use machine learning to classify selected events that violate the safety of the physical layer, such as manipulation or temporary disconnection of connectors. The results show the resting state can be accurately distinguished from selected security breaches for a fiber route subjected to environmental disturbances, where individual events can be classified with nearly 99% accuracy

Anti-VEGF-A affects the angiogenic properties of tumor-derived microparticles.

Tumor derived microparticles (TMPs) have recently been shown to contribute to tumor re-growth partially by inducing the mobilization and tumor homing of specific bone marrow derived pro-angiogenic cells (BMDCs). Since antiangiogenic drugs block proangiogenic BMDC mobilization and tumor homing, we asked whether TMPs from cells exposed to an antiangiogenic drug may affect BMDC activity and trafficking. Here we show that the level of VEGF-A is reduced in TMPs from EMT/6 breast carcinoma cells exposed to the anti-VEGF-A antibody, B20. Consequently, these TMPs exhibit reduced angiogenic potential as evaluated by a Matrigel plug and Boyden chamber assays. Consistently, BMDC mobilization, tumor angiogenesis, microvessel density and BMDC-colonization in growing tumors are reduced in mice inoculated with TMPs from B20-exposed cells as compared to mice inoculated with control TMPs. Collectively, our results suggest that the neutralization of VEGF-A in cultured tumor cells can block TMP-induced BMDC mobilization and colonization of tumors and hence provide another mechanism of action by which antiangiogenic drugs act to inhibit tumor growth and angiogenesis

Alternative spectral window for precise time fiber based transport

Precise time and also stable frequency transfers are approaching the state to become almost standard applications for optical fiber networks, especially they are becoming supported in multiple research and education networks. In principle, the time and frequency transfers are not possible to be implemented over data transport layer, which hasn't been was designed for essential stability, and thus dedicated wave band is required for such implementation. To achieve even better stability for precise time and stable frequency, the bidirectional transmission in single physical medium (fibers and ideally all components along the route) is preferred. The White Rabbit system combines synchronous Ethernet and Precise Time Protocol (IEEE-1588) and it has been designed in the CERN with the aim to provide time synchronization among large number of sensors, actuators and other devices utilized in experiments. Primary aim was the operation over single fiber (to avoid nonreciprocal changes in directions) using standard telecommunication transceivers designed for single fiber operation with reach limited to about 40 km. In this paper, we experimentally verify and compare stability of White Rabbit system operation over long distances up to 300 km using active optical amplification on wavelengths which should not be utilized within spectrum needed for regular data transmissions.</p

Representative flow cytometry plots of viable CEPs, hemangiocytes, and myeloid derived suppressor cells.

<p>An example of the analysis of flow cytometry data obtained from peripheral blood of BALB/c mice is presented. Viable CEPs are determined as (a) positive for VEGFR2 and negative for CD45 as well as (b) positive for CD117 and negative for 7-AAD. Hemangiocytes are determined as (c) positive for CD45 and CXCR4 as well as (d) positive for VEGFR1. Myeloid derived suppressor cells (MDSCs) are determined as positive for (e) both Gr-1 and CD11b.</p

TMPs from cells exposed to anti-VEGF-A antibody exhibit reduced ability to promote endothelial cell activity.

<p>Matrigel plugs containing an equal number of TMPs (0.5×10⁶) from untreated or B20-exposed EMT/6 cells were implanted into the flanks of 8–10 week old BALB/c mice. Matrigel plugs containing PBS were used as a negative control. Ten days later, plugs were removed and then sectioned. (A) Slides were stained with H&E or immunostained with the endothelial cell marker CD31 (designated in red) (scale bar = 100 µm). (B) Microvessel density in the plugs was calculated by counting vessel structures. (C–D) An equal number of TMPs (5×10⁶) from untreated or B20-exposed EMT/6 cells were tested for HUVEC migration (C) and invasion (D) using the modified Boyden chamber assay. PBS was used as a negative control. Cells invading the membrane of the Boyden chamber were stained with Crystal Violet and images were captured using a Leica CTR 6000 microscope. The number of cells invading the membrane were counted and plotted (n>8/group). (E) Aortic rings from BALB/c mice (n = 4/group) were cultured in medium containing 0.1×106 TMPs from untreated or B20-exposed EMT/6 cells. Endothelial cell medium (ECGS) was used as a positive control. Images were captured using an inverted light microscope system (Leica CTR 6000 system) (Scale bar = 500 µm). *, 0.05p>0.001; ***, p<0.001.</p

TMPs from cells exposed to anti-VEGF-A antibody do not induce viable CEP and hemangiocyte mobilization.

<p>(A) An equal number of TMPs (0.5×10⁶) from untreated (CONT) or B20-exposed EMT/6 cells was injected into the tail vein of 8–10 week old non-tumor bearing BALB/c mice (n = 4 mice/group). Control mice were injected with PBS (PBS). One hour later, blood was drawn from the retro-orbital sinus for the evaluation of viable CEPs (CD45−/VEGFR2+/CD117+/7AAD−), MDSCs (Gr1+/CD11b+), and hemangiocytes (CD11b+/CXCR4+/VEGFR1+) using flow cytometry. (B) Half a million TMPs from untreated (CONT) or B20-exposed cells were tagged with PKH26, and subsequently injected into the tail vein of BALB/c mice (n = 4 mice/group). Control mice were injected with PBS. One hour later, blood was drawn by cardiac puncture and total BMDCs (CD45+), viable CEPs, hemagiocytes, and MDSCs were analyzed by flow cytometry. The percentage of the different cell types positive for tagged TMPs was plotted. **, 0.01>p>0.001; ***, p<0.001.</p

TMPs from cells exposed anti-VEGF-A antibody do not promote angiogenesis in tumors.

<p>(A) Matrigel plugs containing an equal number of TMPs (0.5×10⁶) from untreated or B20-exposed EMT/6 cells were implanted into the flanks of 8–10 week old BALB/c mice. Matrigel plugs containing PBS were used as a negative control. Ten days later, plugs were removed and prepared as single cell suspensions. The extracted cells were immunostained for endothelial cells, hemangiocytes and MDSCs and analyzed by flow cytometry. Results are presented as the number of cells per 1 mg Matrigel. (B–E) Eight to ten week old BALB/c mice (n = 4 mice/group) were implanted with 0.5×10⁶ EMT/6 cells into the flanks. When tumors reached a size of approximately 50 mm³, injections with 0.5×10⁶ TMPs from untreated or B20-exposed EMT/6 cells were performed twice weekly. Control mice were injected with PBS. (B) Tumor growth was assessed by a Vernier caliper using the formula, width²×length×0.5. Tumors were removed at endpoint, and subsequently were either (C) stained for CD31 (in red), CD45 (in green), and Hoechst (in blue) for the evaluation of (D) microvessel density and perfusion (scale bar = 100 µm), or (E) prepared as single cell suspensions for the evaluation of MDSCs and hemangiocytes colonization of tumors using flow cytometry. **, 0.01>p>0.001; ***, p<0.001.</p

Exposing tumor cells to anti-VEGF-A antibodies reduces the level of VEGF-A in TMPs without affecting the number of TMPs.

<p>(A) A representative flow cytometry dotplot for TMP quantification. TMPs are approximately 1 µm, and counting beads are 7.35 µm. The number of TMPs per sample was calculated as the ratio between the number of events collected in the counting beads gate and the number of events collected in the TMPs gate over the total number of counting beads loaded in the sample. (B) EMT/6, 4T1 and MDA-MB231 breast carcinoma cells were either left untreated or exposed to 2 µg/ml B20 antibody for 48 h. TMPs were purified from conditioned medium and quantified by flow cytometry. Shown are the means ± S.D. of triplicates. (C) An equal number of TMPs (100,000) from untreated or B-20-exposed EMT/6, 4T1 and MDA-MB231 breast carcinoma cells were used to quantify the level of VEGF-A by ELISA. In some experiments control for B20 antibodies was used in a form of IgG in culture. Shown are the means ± S.D. of triplicates. **, 0.01>p>0.001.</p