p53 dependent senescence by pterostilbene in precancerous bronchial cell.

<p>(A) Induced senescence on HBECR and HBECR/p53i cells by pterostilbene. The cells on glass cover slips were treated with pterostilbene (PT) for 72hr and then stained in situ for senescence-associated β–galactosidase. The representative images are shown (top left). The cells cultured were also treated with PT for 72hr and then collected for C₁₂FDG staining followed by analysis with flow cytometry. (bottom left). The SA-βgal positive cells were quantified by microscopically or flow cytometry. <i>*</i>, <i>P < 0</i>.<i>05; **</i>, <i>P< 0</i>.<i>01</i> by Student’s <i>t</i> test. (B) Change of p21 and p53 by 72hr treatment of PT (5μM).</p

ATM/CHK/p53 Pathway Dependent Chemopreventive and Therapeutic Activity on Lung Cancer by Pterostilbene

<div><p>Among the many stilbenoids found in a variety of berries, resveratrol and pterostilbene are of particular interest given their potential for use in cancer therapeutics and prevention. We purified four stilbenoids from <i>R</i>. <i>undulatum</i> and found that pterostilbene inhibits cancer cell proliferation more efficiently than rhapontigenin, piceatannol and resveratrol. To investigate the underlying mechanism of this superior action of pterostilbene on cancer cells, we utilized a reverse-phase protein array followed by bioinformatic analysis and found that the ATM/CHK pathway is modified by pterostilbene in a lung cancer cell line. Given that ATM/CHK signaling requires p53 for its biological effects, we hypothesized that p53 is required for the anticancer effect of pterostilbene. To test this hypothesis, we used two molecularly defined precancerous human bronchial epithelial cell lines, HBECR and HBECR/p53i, with normal p53 and suppressed p53 expression, respectively, to represent premalignant states of squamous lung carcinogenesis. Pterostilbene inhibited the cell cycle more efficiently in HBECR cells compared to HBECR/p53i cells, suggesting that the presence of p53 is required for the action of pterostilbene. Pterostilbene also activated ATM and CHK1/2, which are upstream of p53, in both cell lines, though pterostilbene-induced senescence was dependent on the presence of p53. Finally, pterostilbene more effectively inhibited p53-dependent cell proliferation compared to the other three stilbenoids. These results strongly support the potential chemopreventive effect of pterostilbene on p53-positive cells during early carcinogenesis.</p></div

Enhanced cytotoxic effects of pterostilbene in precancerous cell lines, HBECR and HBECR/p53i.

<p>(A) Western blotting analysis showing the presence and absence of p53 in HBECR and HBECR/p53i cell lines. (B) Cell viability of HBECR and HBECR/p53i cells (2×10³) treated with different concentrations of pterostilbene ranging from 0 to 100 μM. (C) Cell proliferation ELISA assay with BrdU incorporation. (D) HBECR and HBECR/p53i cell lines were synchronized by double thymidine block. The HBECR cells were only partly synchronized. Cells were released into S phase along with treatment of pterostilbene at the final concentration of 3 μM. At the indicated time point from the thymidine block, cells were harvested and fixed for cell cycle analysis.</p

Activation of ATR-chk1/2-p53 pathway induced by pterostilbene in pre-malignant lung epithelial cell lines.

<p>(A) Cultured HBECR and HBECR/p53i cells were treated with different concentrations of pterostilbene ranging from 0 to 5 μM. After 24 hours, cell lysates were prepared and the proteins were visualized by Western blotting using specific antibodies as indicated. (B) HBECR and HBECR/p53i cells were treated with indicated concentration of ATM inhibitor caffeine and/or pterostilbene for 24 hr. (C) HBECR and HBECR/p53i were treated with ATM activator hydroxyurea (HU) and/or pterostilbene (PT) for 24hr. γH2AX and 53BP1 were detected by immunofluorescence analyses. The images were taken at 63X with confocal microscope. The wide view of the images are in the supplementary figure (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0162335#pone.0162335.s002" target="_blank">S2 Fig</a>). (D) Phosphorylation of H2AX upon HU (hydroxyurea) or PT was determined by Western blot analysis.</p

Effect of pterostilbene on cancer cell proliferation.

<p>(A) Chemical structures of 4 stilbenoids used. (B) A549 cells were seeded in 96-well cell plate. The cells were treated with resveratrol, pterostilbene, rhapontigenin and piceatannol at the final concentration of 1 μM and 5 μM in the presence and absence of FBS. After three days, the cell viability was determined by total live cell counting. (C) A549 cells seeded in 96 well plates were treated with different concentrations of pterostilbene ranging from 0 to 30 μM. After three days, the cell viability was determined by total live cell counting. <i>*</i>, <i>P < 0</i>.<i>05; **</i>, <i>P< 0</i>.<i>01</i> by Student’s <i>t</i> test.</p

Pterostilbene activated ATR/CHK1/p53 pathway to arrests the cell cycle in A549.

<p>(A) Hierarchical clustering of RPPA data from A549 cells treated with pterostilbene at the concentration of 2.5 μM and 5 μM for 48 hours in serum free media (n = 4 for each samples). Protein expression levels of A549 without pterostilbene treatment were used as a control. The data are presented in a matrix format: rows represent individual protein features (186 proteins), and columns represent individual samples. Each cell in the matrix represents the expression level of a protein feature in an individual cell sample (left). A total of 16 proteins showing significant differences with <i>P</i> < 0.05 by Student’s <i>t</i> test are shown in the right panel. (B) Western blotting results of A549 cells treated with for the molecules implicated in DNA damage responses and cell cycle regulations.</p

p53 dependent effects of pterostilbene in precancerous bronchial cell senescence.

<p>The p53 dependent cytotoxic effects of resveratrol, pterostilbene, rhapontigenin and piceatannol on precancerous bronchio-epithelial cells, with or without functional p53. <i>*</i>, <i>P < 0</i>.<i>05; **</i>, <i>P< 0</i>.<i>01</i> by Student’s <i>t</i> test.</p

Effect of pterostilbene treatment on the cell cycle profile.

<p>(A) A549 cells in 60mm dishes cultured with or without FBS were treated with indicated concentrations of pterostilbene for 48hr and the cell cycle was analyzed. (B) For synchronization at G0 phase, A549 cells were treated 2mM thymidine for 18hr and released media without thymidine for 6hr. For second block, the cell were treated 2mM thymidine for 18hours and released indicated time with or without pterostilbene and the cell cycle was measured. The red areas represents G0/G1 or G2/M stage cells and the blue shaded area represents S-stage cells after analyzed by Modfit LT program from Verity Software (Topsham, ME).</p

StMary-PPGRR.zip

StMary Introduction The StMary dataset is Photoplethysmography(PPG) and Respiratory Rate(RR) dataset acquired from surgical intensive care unit(SICU) of Catholic University Seoul St. Mary's Hospital, South Korea. Data Collection The StMary dataset was acquired from a SICU at the hospital using our in-house web application, PPG Signal Collector. Using laptop, we connected a network between the web application and a patient monitor. Through pulse oximeter which connected to the patient monitor, we collected photoplethysmogram of patients and healthy volunteers. Subject's demographic information must be entered before collecting. After that, PPG signal continuously recorded during the time limit we had set and Respiraotry Rate also recorded manually by medical staff pressing spacebar at the exhalation time. The time limit was set up to 2 minutes and 10 seconds. When the time ends, the data is recorded as history and has stored in the database. The collection procedure is as follows:  Move into local monitoring page through the browser installed in the laptop. (Set up the measuring time additionally)  Register subject’s demographic – gender, age, diagnosis – to start recording.   If a window pop-up, click the start button to record PPG.   Press spacebar when the subject exhales.   When the time outs, the window closed, and the data stored automatically.  Data Files The StMary dataset consists of 50 patients and 50 healthy volunteers both aged 18 and older. The dataset has a demographic information, PPG signal and exhalation timestamp for each de-identified subject. 2-minute PPG was collected for each subject twice, and it was sampled into 125Hz. A unique ID is generated for each collected 2-minute PPG. While this ID serves as an identifier for individual data, serving as a de-identifier for the subject at the same time. Patient.csv Diagnosis: Indicates diagnosis of the subject. For volunteers, mark '0'. For patients, name the diagnosis. Gender: Indicates gender of the subject. ("M"/"F") Age: Indicates age of the subject. startMeasureTime: Indicates time when the data starts to move into DB server. endMeasureTime: Measurement ending time. Pleth.csv Timestamp: Indicates time when the PPG at that time collected. Pleth: Indicates PPG which acquired from pulse oximeter.The StMary dataset is Photoplethysmography(PPG) and Respiratory Rate(RR) dataset acquired from surgical intensive care unit(SICU) of Catholic University Seoul St. Mary's Hospital, South Korea. respirationTimeline.csv Timestamp: Indicates time when subject exhales. All protocol followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000. All data  were anonymized, deidentified and aggregated. This data  was approved by the Institutional Review Board of the Ethics Committee of Seoul St. Mary’s Hospital (IRB No. KC21ONSI0839) and the need for consent was waived by IRB. </p

Proactive strategy for long-term biological research aimed at low-dose radiation risk in Korea

<p><b>Purpose:</b> Since the 2011 Fukushima nuclear power plant accident, Korean radiation experts have agreed that reliable data on health risks of low-dose radiation (LDR) are needed to ease the anxiety of lay people. The intent of this study was to devise a sustainable biological program suited for the research environment in Korea and aimed at the health effects of radiation exposures <100 millisieverts (mSv). To address pressing public concerns over LDR risk, we investigated the current understanding of LDR effects by analyzing the previous reports of international authorities for radiation protection and research publications that appeared after the Chernobyl accident. A research program appropriate for societal and scientific inclinations of Korea was then devised based on input from Korean radiation scientists.</p> <p><b>Conclusions:</b> After review by our advisory committee, program priorities were set, calling for an agenda that focused on dose-response relationships in carcinogenesis, health span responses to lifestyle variations, and systemic metabolic changes. Our long-term biological research program may contribute scientific evidence to reduce the uncertainties of LDR health risks and help stakeholders formulate policies for radiation protection.</p