Calreticulin exposure on malignant blasts predicts a cellular anticancer immune response in patients with acute myeloid leukemia

Experiments performed in mice revealed that anthracyclines stimulate immunogenic cell death that is characterized by the pre-apoptotic exposure of calreticulin (CRT) on the surface of dying tumor cells. Here, we determined whether CRT exposure at the cell surface (ecto-CRT) occurs in human cancer in response to anthracyclines in vivo, focusing on acute myeloid leukemia (AML), which is currently treated with a combination of aracytine and anthracyclines. Most of the patients benefit from the induction chemotherapy but relapse within 1–12 months. In this study, we investigated ecto-CRT expression on malignant blasts before and after induction chemotherapy. We observed that leukemic cells from some patients exhibited ecto-CRT regardless of chemotherapy and that this parameter was not modulated by in vivo chemotherapy. Ecto-CRT correlated with the presence of phosphorylated eIF2α within the blasts, in line with the possibility that CRT exposure results from an endoplasmic reticulum stress response. Importantly, high levels of ecto-CRT on malignant myeloblasts positively correlated with the ability of autologous T cells to secrete interferon-γ on stimulation with blast-derived dendritic cell. We conclude that the presence of ecto-CRT on leukemia cells facilitates cellular anticancer immune responses in AML patients.     

Molecular mechanisms that produce secondary MDS/AML by RUNX1/AML1 point mutations

RUNX1/AML1 point mutations have been identified in myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) patients. A heterozygous germline mutation of the RUNX1 gene causes a familial platelet disorder with a predisposition to AML. RUNX1 mutations have also been detected with high frequency in minimally differentiated AML M0 subtypes and myelodysplastic/myeloproliferative neoplasms. Here we propose a new disease category of myelodysplastic neoplasms (MDN) consisting of MDS refractory anemia with excess blasts and AML with myelodysplasia-related changes, including therapy-related cases. RUNX1 mutations have been detected in about 20% of patients with "MDN". Among the MDN cases, histories of radiation exposure, therapy-related myeloid neoplasms after successful treatment for acute promyelocytic leukemia, and leukemic transformation of myeloproliferative neoplasms have been reported to have a strong association with RUNX1 mutations. The mutations occur in a normal, a receptive, or a disease-committed hematopoietic stem cell. It is suspected that the "MDN" phenotypes are defined by the RUNX1 mutations in addition to some other abnormalities.     

Combining near‐ and far‐field exposure for an organ‐specific and whole‐body RF‐EMF proxy for epidemiological research: A reference case

A framework for the combination of near‐field (NF) and far‐field (FF) radio frequency electromagnetic exposure sources to the average organ and whole‐body specific absorption rates (SARs) is presented. As a reference case, values based on numerically derived SARs for whole‐body and individual organs and tissues are combined with realistic exposure data, which have been collected using personal exposure meters during the Swiss Qualifex study. The framework presented can be applied to any study region where exposure data is collected by appropriate measurement equipment. Based on results derived from the data for the region of Basel, Switzerland, the relative importance of NF and FF sources to the personal exposure is examined for three different study groups. The results show that a 24‐h whole‐body averaged exposure of a typical mobile phone user is dominated by the use of his or her own mobile phone when a Global System for Mobile Communications (GSM) 900 or GSM 1800 phone is used. If only Universal Mobile Telecommunications System (UMTS) phones are used, the user would experience a lower exposure level on average caused by the lower average output power of UMTS phones. Data presented clearly indicate the necessity of collecting band‐selective exposure data in epidemiological studies related to electromagnetic fields. Bioelectromagnetics 34:366–374, 2013. © 2012 Wiley Periodicals, Inc.     

Spatial and temporal RF electromagnetic field exposure of children and adults in indoor micro environments in Belgium and Greece

Personal radio frequency electromagnetic field (RF-EMF) exposure, or exposimetry, is gaining importance in the bioelectromagnetics community but only limited data on personal exposure is available in indoor areas, namely schools, crèches, homes, and offices. Most studies are focused on adult exposure, whereas indoor microenvironments, where children are exposed, are usually not considered. A method to assess spatial and temporal indoor exposure of children and adults is proposed without involving the subjects themselves. Moreover, maximal possible daily exposure is estimated by combining instantaneous spatial and temporal exposure. In Belgium and Greece, the exposure is measured at 153 positions spread over 55 indoor microenvironments with spectral equipment. In addition, personal exposimeters (measuring EMFs of people during their daily activities) captured the temporal exposure variations during several days up to one week at 98 positions. The data were analyzed using the robust regression on order statistics (ROS) method to account for data below the detection limit. All instantaneous and maximal exposures satisfied international exposure limits and were of the same order of magnitude in Greece and Belgium. Mobile telecommunications and radio broadcasting (FM) were most present. In Belgium, digital cordless phone (DECT) exposure was present for at least 75% in the indoor microenvironments except for schools. Temporal variations of the exposure were mainly due to variations of mobile telecommunication signals. The exposure was higher during daytime than at night due to the increased voice and data traffic on the networks. Total exposure varied the most in Belgian crèches (39.3%) and Greek homes (58.2%).     

Cell surface marker phenotypes and gene expression profiles of murine radiation-induced acute myeloid leukemia stem cells are similar to those of common myeloid progenitors

Radiation exposure induces acute myeloid leukemia (AML) in humans and mice. Recent studies postulated that AML stem cells of spontaneous human AML arise from hematopoietic stem cells. However, other studies support the possibility that short-lived committed progenitors transform into AML stem cells, accompanied by a particular gene mutation. It remains unclear whether AML stem cells are present in radiation-induced AML, and information regarding AML-initiating cells is lacking. In this study, we identified and analyzed AML stem cells of mice with radiation-induced AML. The AML stem cells were identified by transplanting 100 bone marrow cells from mice with radiation-induced AML. We injected 100 cells of each of seven cell populations corresponding to different stages of hematopoietic cell differentiation and compared the latencies of AMLs induced in recipient mice. The identified radiation-induced AML stem cells frequently displayed similarities in both CD antigen and gene expression profiles with normal common myeloid progenitors. The number of common myeloid progenitor-like AML stem cells was significantly increased in mice with radiation-induced AML, but the progeny of common myeloid progenitors was decreased. In addition, analysis of radiation effects on the hematopoietic system showed that common myeloid progenitor cells were extremely radiosensitive and that their numbers remained at low levels for more than 2?months after radiation exposure. Our results suggest that murine radiation-induced AML stem cells arise from radiosensitive cells at a common myeloid progenitor stage.     

ATPR triggers acute myeloid leukaemia cells differentiation and cycle arrest via the RARα/LDHB/ERK‐glycolysis signalling axis

Acute myeloid leukaemia (AML) remains a therapeutic challenge and improvements in chemotherapy are needed. 4‐Amino‐2‐trifluoromethyl‐phenyl retinate (ATPR), a novel all‐trans retinoic acid (ATRA) derivative designed and synthesized by our team, has been proven to show superior anticancer effect compared with ATRA on various cancers. However, its potential effect on AML remains largely unknown. Lactate dehydrogenase B (LDHB) is the key glycolytic enzyme that catalyses the interconversion between pyruvate and lactate. Currently, little is known about the role of LDHB in AML. In this study, we found that ATPR showed antileukaemic effects with RARα dependent in AML cells. LDHB was aberrantly overexpressed in human AML peripheral blood mononuclear cell (PBMC) and AML cell lines. A lentiviral vector expressing LDHB‐targeting shRNA was constructed to generate a stable AML cells with low expression of LDHB. The effect of LDHB knockdown on differentiation and cycle arrest of AML cells was assessed in vitro and vivo, including involvement of Raf/MEK/ERK signalling. Finally, these data suggested that ATPR showed antileukaemic effects by RARα/LDHB/ ERK‐glycolysis signalling axis. Further studies should focus on the underlying leukaemia‐promoting mechanisms and investigate LDHB as a therapeutic target.     

Specific absorption rate and electric field measurements in the near field of six mobile phone base station antennas

In this article, the exposure to radio frequency electromagnetic fields was studied in close proximity (distances of 10, 100, 300, and 600 mm) to six base station antennas. The specific absorption rate (SAR) in 800 mm × 500 mm × 200 mm box phantom as well as unperturbed electric field (E) in air was measured. The results were used to determine whether the measurement of local maximum of unperturbed electric field can be used as a compliance check for local exposure. Also, the conservativeness of this assessment method compared to the ICNIRP basic restriction was studied. Moreover, the assessment of whole‐body exposure was discussed and the distance ranges presented in which the ICNIRP limit for local exposure could be exceeded before the limit for whole‐body SAR. These results show that the electric field measurement alone can be used for easy compliance check for the local exposure at all distances and for all antenna types studied. However, in some cases when the local peak value of E was compared directly to the ICNIRP reference level for unperturbed E, the exposure was overestimated only very slightly (by factor 1.1) compared to the basic restriction for localized SAR in a human, and hence these results can not be generalized to all antenna types. Moreover, it was shown that the limit for localized exposure could be exceeded before the limit for the whole‐body average SAR, if the distance to the antenna was less than 240 mm. Bioelectromagnetics 30:307–312, 2009. © 2009 Wiley‐Liss, Inc.     

Electromagnetic fields produced by GSM cellular phones and heart rate variability

In this study, 26 healthy young volunteers were submitted to 900 MHz (2 W) GSM cellular phone exposure and to sham exposure in separate sessions. The study was designed to assess cardiac regulatory mechanism in different autonomic nervous system (ANS) states during exposure to low-intensity EMF. Rest-to-stand protocol was applied to evaluate ANS in quiet condition (rest, vagal prevalence) and after a sympathetic activation (stand). The procedure is conducted twice in a double-blind design: once with a genuine EMF exposure and once with a sham exposure (at least 24 h apart). During each session three-leads electrocardiograms were recorded and RR series extracted off-line. Time domain and frequency domain HRV parameters were calculated in every phase of the protocol and during different exposures. The analysis of the data show there was no statistically significant effect due to EMF exposure both on main (i.e., RR mean) and most of the other HRV parameters. A weak interaction between some HRV parameters (i.e., SDNN, TINN, and triangular index in time domain and LF power in frequency domain analysis) and RF exposure was observed and this effect seems to be gathered around the sympathetic response to stand.     

Exposure assessment of mobile phone base station radiation in an outdoor environment using sequential surrogate modeling

Human exposure to background radiofrequency electromagnetic fields (RF‐EMF) has been increasing with the introduction of new technologies. There is a definite need for the quantification of RF‐EMF exposure but a robust exposure assessment is not yet possible, mainly due to the lack of a fast and efficient measurement procedure. In this article, a new procedure is proposed for accurately mapping the exposure to base station radiation in an outdoor environment based on surrogate modeling and sequential design, an entirely new approach in the domain of dosimetry for human RF exposure. We tested our procedure in an urban area of about 0.04 km² for Global System for Mobile Communications (GSM) technology at 900 MHz (GSM900) using a personal exposimeter. Fifty measurement locations were sufficient to obtain a coarse street exposure map, locating regions of high and low exposure; 70 measurement locations were sufficient to characterize the electric field distribution in the area and build an accurate predictive interpolation model. Hence, accurate GSM900 downlink outdoor exposure maps (for use in, e.g., governmental risk communication and epidemiological studies) are developed by combining the proven efficiency of sequential design with the speed of exposimeter measurements and their ease of handling. Bioelectromagnetics 34:300–311, 2013. © 2012 Wiley Periodicals, Inc.     

Targeting miR‐193a‐AML1‐ETO‐β‐catenin axis by melatonin suppresses the self‐renewal of leukaemia stem cells in leukaemia with t (8;21) translocation

AML1‐ETO, the most common fusion oncoprotein by t (8;21) in acute myeloid leukaemia (AML), enhances hematopoietic self‐renewal and leukemogenesis. However, currently no specific therapies have been reported for t (8;21) AML patients as AML1‐ETO is still intractable as a pharmacological target. For this purpose, leukaemia cells and AML1‐ETO‐induced murine leukaemia model were used to investigate the degradation of AML1‐ETO by melatonin (MLT), synthesized and secreted by the pineal gland. MLT remarkedly decreased AML1‐ETO protein in leukemic cells. Meanwhile, MLT induced apoptosis, decreased proliferation and reduced colony formation. Furthermore, MLT reduced the expansion of human leukemic cells and extended the overall survival in U937T‐AML1‐ETO‐xenografted NSG mice. Most importantly, MLT reduced the infiltration of leukaemia blasts, decreased the frequency of leukaemia stem cells (LSCs) and prolonged the overall survival in AML1‐ETO‐induced murine leukaemia. Mechanistically, MLT increased the expression of miR‐193a, which inhibited AML1‐ETO expression via targeting its putative binding sites. Furthermore, MLT decreased the expression of β‐catenin, which is required for the self‐renewal of LSC and is the downstream of AML1‐ETO. Thus, MLT presents anti‐self‐renewal of LSC through miR‐193a‐AML1‐ETO‐β‐catenin axis. In conclusion, MLT might be a potential treatment for t (8;21) leukaemia by targeting AML1‐ETO oncoprotein.     

The adaptive response modifies latency for radiation-induced myeloid leukemia in CBA/H mice.

We have investigated the effect of the adaptive response on acute myeloid leukemia (AML) induced in CBA/Harwell mice by a chronic radiation exposure. Groups of mice irradiated with a total dose of 1. 0 Gy at two different chronic dose rates (0.5, 0.004 Gy/h) had similar frequencies of AML. Compared to control animals that did not develop AML, irradiation at either of these dose rates did not change the longevity of the mice that did not die of leukemia. The survival rates of irradiated mice that did develop leukemia in the two groups were not different from each other, indicating that the dose rates produced similar responses and therefore were both chronic exposures. We then tested the ability of a chronic 10-cGy (0. 5 Gy/h) exposure to ionizing radiation, mild hyperthermia (40.5 degrees C whole-body, 60 min) or treatment with interleukin-1 (1500 U i.p.) to induce an adaptive response and modify the frequency or latency of AML which resulted from a subsequent (24 h later) 1.0-Gy (0.5 Gy/h) chronic radiation exposure. The frequency of radiation-induced leukemia was not changed in mice given any of the three adapting treatments 24 h prior to the chronic 1.0-Gy dose that induced leukemia. However, the latent period for development of AML was significantly increased by both the prior low radiation dose and mild hyperthermia treatment. Injection of interleukin-1, in contrast, may have reduced the latent period. Similar to the single 1.0-Gy chronic exposure alone, none of the adapting treatments prior to that exposure influenced the survival of animals that did not develop AML. These results indicate that an earlier exposure to a small adapting dose of radiation or to a mild heat stress can influence secondary steps in radiation-induced carcinogenesis.     

Effects of 50 Hz magnetic field on cell cycle kinetics and the colony forming ability of budding yeast exposed to ultraviolet radiation.

To investigate the effects of extremely low frequency magnetic fields on ultraviolet radiation (UV) exposed budding yeast, haploid yeast (Saccharomyces cerevisiae) cells of the strain SEy2101a were exposed to 50 Hz sine wave magnetic field (MF) of 120 microT with simultaneous exposure to UV radiation. Most of the UV energy was in the UVB range (280-320 nm). The biologically weighted (CIE action spectrum) dose level for the UV radiation was 175 J/m2. We examined whether 50 Hz MF affected the ability of UV irradiated yeast cells to form colonies (Colony Forming Units, CFUs). In addition, the effect of coexposure on cell cycle kinetics was investigated. Although the significant effect of MF on the cell cycle phases of UV exposed yeast cells was seen only at one time point, the overall results showed that MF exposure may influence the cell cycle kinetics at the first cycle after UV irradiation. The effect of our particular MF exposure on the colony forming ability of the UV irradiated yeast cells was statistically significant 420 min after UV irradiation. Moreover, at 240, 360, and 420 min after UV irradiation, there were fewer CFUs in every experiment in (UV+MF) exposed populations than in only UV exposed yeast populations. These results could indicate that MF exposure in conjunction with UV may have some effects on yeast cell survival or growth.     

Relative contribution of residential and occupational magnetic field exposure over twenty-four hours among people living close to and far from a power line

This study sought to estimate the relative contribution of exposure to 50 Hz magnetic fields experienced at home, at work/school, or elsewhere to the total exposure over 24 hr. Personal exposure meters were carried by 97 adults and children in the Stockholm area. About half of the subjects lived close (<50 m) to a transmission line and half far (>100 m) away. Spot measurements and calculations for the residential exposure were also made. For subjects living<50 m from the line, the exposure at home contributed about 80% of the total magnetic field exposure, measured in mT-hours. Adults living far away experienced only 38% of the total exposure at home, but children still received 55%. Subjects with low time-weighted average (TWA) exposure both at home and at work spent 84% of their time in fields <0.1 microT, and those with high TWA at both locations spent 69% of their time in fields > or = 0.2 microT. This contrast was diluted if only exposure at one location was considered. For spot measurements and calculations of the residential exposure, both sensitivity and specificity was good. However, the intermediate field exposure category (0.1-0.19 microT) showed poor correlation to the 24 hr personal measurements.     

Analysis of individual- and school-level clustering of power frequency magnetic fields

This study reports the continuous 8-h monitoring of data on extremely low-frequency magnetic fields (ELF-MF) relating to 14 children and 35 teachers in 11 elementary schools in Northern Taiwan. It was anticipated that the subjects in two of these campuses would have elevated exposure to ELF-MF as a result of their close proximity to high-voltage (161 kilo-Volt, kV) power lines. The results of our analysis reveal that in those schools with high-voltage power lines running through the campuses, the mean ELF-MF exposure level (0.38 +/- 0.51 micro-Tesla (microT), or 0.15, 0.25 and 0.44 microT at the respective 25th, 50th and 75th percentiles) was higher than the mean ELF-MF exposure level for campuses situated far away from such high-voltage power lines (0.14 +/- 0.27 microT, or 0.04, 0.06 and 0.10 microT at the respective 25th, 50th and 75th percentiles). The multi-level analytical technique, which takes individual measurements as the analytical unit, and which also takes into consideration the inter-correlation between measurements from the same individual and/or campus, was also applied to the analysis of the data. We conclude that individual-level and school-level clustering of the measurements, both of which were discernible in this study, should be taken into consideration in any future analysis of data obtained from the continuous monitoring of exposure to ELF-MF.     

Effect of microwave radiation on human EEG at two different levels of exposure

This study is aimed at evaluating the effect of microwave radiation on human brain bioelectric activity at different levels of exposure. For this purpose, 450 MHz microwave exposure modulated at 40 Hz frequency was applied to a group of 15 healthy volunteers at two different specific absorption rate (SAR) levels: a higher level of 0.303 W/kg (field strength 24.5 V/m) and a lower level of 0.003 W/kg (field strength 2.45 V/m). Ten exposure cycles (1 min off and 1 min on) at fixed SAR values were applied. A resting eyes‐closed electroencephalogram (EEG) was continuously recorded. Results showed a statistically significant increase in the EEG power in the EEG beta2 (157%), beta1 (61%) and alpha (68%) frequency bands at the higher SAR level, and in the beta2 (39%) frequency band at the lower SAR level. Statistically significant changes were detected for six individual subjects in the EEG alpha band and four subjects in the beta1 and beta2 bands at the higher SAR level; three subjects were affected in the alpha, beta1 and beta2 bands at the lower SAR level. The study showed that decreasing the SAR 100 times reduced the related changes in the EEG three to six times and the number of affected subjects, but did not exclude the effect. Bioelectromagnetics 34:264–274, 2013. © 2012 Wiley Periodicals, Inc.     

High-Throughput Exposure Assessment Tool (HEAT) for exposure-based prioritization of chemicals

Abstract

High-throughput methods are now routinely used to rapidly screen chemicals for potential hazard. However, hazard-based decision-making excludes important exposure considerations resulting in an incomplete estimation of chemical safety. Models to estimate exposure exist, but are generally unsuited to keep up with high-throughput demands. The High-Throughput Exposure Assessment Tool (HEAT) is designed to efficiently predict near-field exposure to consumers and workers via inhalation, oral and dermal routes. HEAT is based on well-known modeling algorithms and provides default model parameters to support reasonably conservative exposure estimates. Underlying chemical-specific data are uploaded or entered by the end user. HEAT’s main strength is the flexible tiered screening functionality, which enables exposure estimates for single or multiple chemicals simultaneously. Hypothetical case examples highlighting the application of HEAT to more complex exposure estimates for alternative and aggregate assessments are provided.     

Measurement of the individual exposure to 50 and 16 2/3 Hz magnetic fields within the Bavarian population.

This study investigates the individual magnetic field exposures at 16 2/3 and 50 Hz of 1952 people, selected from the Bavarian population. Personal flux density meters ("Field Watcher FW2A") were worn by the participants for 24 h. Every second, the flux density was recorded for both frequencies and for the three spatial axes (dynamic range per axis: several nT up to 100 microT at 50 Hz, 150 microT at 16 2/3 Hz). For 50 Hz fields, the mean of the 1,952 individual means was 0.101 microT and that of the individual medians was 0.047 microT. High level exposures occurred mainly during working hours. Only 2.4% of the subjects showed individual medians higher than 0.2 microT. About 53% of all volunteers were working on the day of recording. Levels for craftsmen (n = 148; mean individual mean: 0.166 microT) were generally higher than those for office workers (n = 624; mean individual mean: 0.107 microT). Flux densities exceeding 100 microT at 50 Hz were measured in 31 persons. The total time with such extreme exposures amounts to nearly 21 min, less than 0.001% of the total time for all measurements (5.3 years). To our knowledge, this is the first exposure study where 16 2/3 Hz magnetic fields (caused by electrified railways) have additionally been monitored over 24 h. For persons living next to railway lines, the mean individual mean (0.156 microT) and mean individual median (0.102 microT) were calculated. Over all, the mean exposures are only 0.1% of the magnetic flux density limit for 50 Hz (100 microT) and about 0.05% of the limit (300 microT) for 16 2/3 Hz recommended by the International Commission on Non-Ionizing Radiation Protection.