Lecture 32, April 18,2016 Chapter 1 Book 2 Assignments • 1.Read chapter 1 of book 2 for Wednesday • 2. Papers Due Friday • 3. Read Chapter 3 of book 2 for Monday Effects of RF and ELF on Cells of the Immune System Tatjana Paunesku and Gayle E. Woloschak Basic Problems in 2006 on Cell Culture and Animal Experiments • 1. No underlying theory to organize a large number of results. • 2. Thus many experiments with different results were the result of the fact that they were different with respect to variables they did not measure or in many cases they did not know were important. • 3. For example, time in cell life cycle, background magnetic field etc. Immune System • 1. Choice of the immune system in part as it senses the environment to protect us therefore we expect it to pick up environmental signals such EMF. • 2. However , since it responds to many stimulus it may be hard to separate the effects of the EMF. Issues • 1. Very large range of exposure parameters Frequency, Power levels, length of pulses, repetition rate , mechanism. • 2. Many different biological material are studied • 3.Do not understand the biology well enough to trace cause and effect from EMF to the end results in health effects. • 4. Short life time of the cells in the immune system. Issues • 5. Different endpoints for the experiments. • 6. Multiple sources of stress. • 7. In vitro vs in vivo. More feedback and repair processes in vivo. Example Human Study vs Mice • 1 Six Human Exposed 8h/d for 5y to 50Hz with B=0.2 to 6.6µT • 2. Results total lymphocytes, CD4+ and CD3+ cell count decreases and NK-Killer cells Increase. • 3. 6 month later , CD4+ and CD3+ and CD19+ cell counts are +28, +22, and =17%, respectively • 4. Another study had contradictory results. In 12 Swiss Mice • 1. Exposed for 109 days to same conditions • 2. Results total lymphocytes, leukocytes, polymorphonuclear neutrophils, and CD4+ cells were significantly lower in exposed animals NK killer cell counts were lower. Other results • 1, Changes in NK cells for workers in the fields generated by induction heater. • 2. Welders had CD4+ and CE8+ cells decreased. • 3. Peripheral Blood Cells Cultured Ex Vivo show changes. CONCLUSION • You are getting change but you do not know what is going on. Animal Studies • 1. 900MHz in TEM Cell to GSM at SAR 1 and 2W/kg saw no short term changes. Some at 1 week and none for 2 or 4 weeks. • 2. Another set of experiments on showed changes some times and not at other times. • 3. Again says something is happening but you do not know how the system works. Chromosome Damage (Sister Chromatid Exchange, Aberrations, etc.) • 1. Study of 24 employees in Norwegian transformer factory high voltage lab showed twice the median number of sister chromatid breaks (5 vs. 2.5 per 50 cells) compared to controls. • 2. Other experiments show no change in damage for exposure to 835.62 MHz for FDMA, CW at 68W yielding power density of 860 W/m2; and the mean SAR was 4.4 or 5.0 W/kg. for 48 and 72hr. DNA Damage as Single-Strand and Double-Strand Breaks • 1. No damage for 2h,2450MHz at 21W and average SAR= 2.13W/kg • 2. Another group for CW at 1.9GHz at 10W/kg for 4 h showed no damage. • 3. Another group at 900MHz 50W SAR to 10W/kg no damage. • 4. Another group shows slowing of the repair processes for 100Hz,1mT and UVB for DNA by 34% Leukemia and Lymphoma • 1. Two conflicting Studies on Leukemia and lymphoma. • A. For example, in 1997 Repacholi and others found increased odds’ ratio (OR) of 2.4, P ¼ 0.006, 95% confidence interval (CI) of 1.3–4.5 for the development of lymphoma in lymphoma-prone E mu-Pim1 transgenic mice exposed to pulsemodulated RF fields twice for 30 min/d for up to 18 months (plane-wave fields of 900 MHz with a pulse repetition frequency of 217 Hz, pulse width of 0.6 ms; incident power densities were 2.6 to 13W/m , and SAR of 0.008 to 4.2W/kg, averaging 0.13– 1.4W/kg) 2 Leukemia and Lymphoma • B. Utteridge and others exposed 120 E mu-Pim1 heterozygous mice and 120 wildtype mice to GSM-modulated 898.4 MHz radiation for 1 h/d, 5 d/week at SARs of 0.25, 1.0, 2.0, and 4.0 W/kg in ‘‘Ferriswheel’’ exposure systems for up to 104 weeks (26 months) and found no increase. Possible Mechanisms of Action of Radiofrequency Radiation and Electromagnetic Field • 1. Direct DNA damage not observed. • 2. Change in Charge Distribution on proteins and protein configuration postulated. • 3. Activation of promoters of c-myc and HSP70 genes. • 4. Changes in ROS, reactive oxygen species Ca2+ Homeostasis • 1 Ca++ is a second messenger. • 2. One study shows no effect of 915MHz 2W/kg on Calcium concentrations and another shows an Increase in activated cells. • 3. ELF exposure show changes at 60Hz and 100µT Nonthermal Heat Shock Effects • 1. Heat Shock Proteins would be better described as stress proteins. • 2. At 895 and 91 5MHz ,5.4 mW/kg changes in chromatin conformation • 3. Belyaev and Alipov Frequency windows for strong effects at 8 and 58Hz at 21µT Gene and Protein Activation • 1. Increased Binding Activity of DNA to NF-kB transcription factor 8.2GHz, 1000 pulses/s, average 50 W/m2; SAR was 10.8 + 7.1W/kg • 2. HL60 cells to a 0.1 mT at 50 Hz increase binding to C-AMP • 3. Exposed to 50 MHz, modulated at 16Hz, 0.2 mT plus 60 V/m leads overexpression of the ets1 mRNA in Jurkat cells Induction of Reactive Oxygen Species • 1. Simko and Mattsson show ELF - EMF exposure increases ROS in macrophages • 2. Zmyslony et.al shows combined effects of 930 MHz CW and FeCl2 at 1.5 W/kg increase ROS • 3. Ding et.al exposed HL-60 cells to 60 Hz, 5 mT and H2O2 (85 or 100 mM) • Showing it as an apoptosis promoter Planaria Data ROS Data On Planaria Conclusions—Possible Explanations of Contradictory Results • 1. Biological systems are time dependent. • 2. There are parallel paths, feedback paths and repair mechanisms. • 3. Basic problem is that the biology is not understood in terms of how the chemistry and dynamics proceed from one stage to the next. • 4. Experiments thought to be the same are not.