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The Role of Temporal Sensing in Bio Electromagnetic Effects

The Role of Temporal Sensing in Bio Electromagnetic Effects

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Published by karlmalatesta
In a series of classical documents that constitute giant milestones in modern bioelectromagnetic research, Ted Litovitz has discovered what might well be the appropriate approach to making cell phone technology if not safe, at least safer. The superposition of extreme low frequency (ELF) noise on the incident wave that directly affects the brain, might hold the key to block or reduce the non-thermal mechanisms that mediate ultra-structural cellular injury. This would alleviate the suffering of millions of patients around the globe suffering from electromagnetic hypersensitivity (electromagnetic sensitivity). Once the human cell is not able to generate these deadly mechanisms, the door is open to turn the microwave syndrome and the radiofrequency disease into a painful memory of an obsolete scientific past. This possibility of confusing cells by electromagnetic noise and thereby preventing the generation of the free radical cascade injury, stands out as a technological instrument that might help us close the cellular windows to different frequencies and magnetic intensities. It is the moral duty of the government of the United States to generate IMMEDIATE research initiatives in this direction in order to protect the people. In front of the explosive epidemic of cases of leukemia (and other malignancies, just to mention a few pathologies) around cell phone base stations and of brain tumors in cell phone users, the variation of field parameters brings hope in the sense that scientific development might one day achieve a safe wireless technology so that our children stop getting sick because of the use of Wi-Fi use in hundreds of thousands of schools and homes around the planet. Please take a look at Ted Litovitz´s conference on “Electromagnetic Fields- The Science on Human Health Effects” at: http://video.google.com/videoplay?docid=7498052433339986964#
In a series of classical documents that constitute giant milestones in modern bioelectromagnetic research, Ted Litovitz has discovered what might well be the appropriate approach to making cell phone technology if not safe, at least safer. The superposition of extreme low frequency (ELF) noise on the incident wave that directly affects the brain, might hold the key to block or reduce the non-thermal mechanisms that mediate ultra-structural cellular injury. This would alleviate the suffering of millions of patients around the globe suffering from electromagnetic hypersensitivity (electromagnetic sensitivity). Once the human cell is not able to generate these deadly mechanisms, the door is open to turn the microwave syndrome and the radiofrequency disease into a painful memory of an obsolete scientific past. This possibility of confusing cells by electromagnetic noise and thereby preventing the generation of the free radical cascade injury, stands out as a technological instrument that might help us close the cellular windows to different frequencies and magnetic intensities. It is the moral duty of the government of the United States to generate IMMEDIATE research initiatives in this direction in order to protect the people. In front of the explosive epidemic of cases of leukemia (and other malignancies, just to mention a few pathologies) around cell phone base stations and of brain tumors in cell phone users, the variation of field parameters brings hope in the sense that scientific development might one day achieve a safe wireless technology so that our children stop getting sick because of the use of Wi-Fi use in hundreds of thousands of schools and homes around the planet. Please take a look at Ted Litovitz´s conference on “Electromagnetic Fields- The Science on Human Health Effects” at: http://video.google.com/videoplay?docid=7498052433339986964#

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Bioelectromagnetics 18:388–395 (1997)
The Role of Temporal Sensing inBioelectromagnetic Effects
T.A. Litovitz,
1
M. Penafiel,
1
D. Krause,
2
D. Zhang,
2
and J.M. Mullins
2
*
1
Department of Physics, The Catholic University of America, Washington, DC
 2
Department of Biology, The Catholic University of America, Washington, DC
Experiments were conducted to see whether the cellular response to electromagnetic (EM) fieldsoccurs through a detection process involving temporal sensing. L929 cells were exposed to 60 Hzmagnetic fields and the enhancement of ornithine decarboxylase (ODC) activity was measured todetermine cellular response to the field. In one set of experiments, the field was turned alternatelyoff and on at intervals of 0.1 to 50 s. For these experiments, field coherence was maintained byeliminating the insertion of random time intervals upon switching. Intervals
°
1 s produced noenhancement of ODC activity, but fields switched at intervals
¢
10 s showed ODC activities thatwere enhanced by a factor of approximately 1.7. These data indicate that it is the interval over whichfield parameters (e.g., amplitude or frequency) remain constant, rather than the interval over whichthe field is coherent, that is critical to cellular response to an EMF. In a second set of experiments,designed to determine how long it would take for cells to detect a change in field parameters, thefield was interrupted for brief intervals (25–200 ms) once each second throughout exposure. In thissituation, the extent of EMF-induced ODC activity depended upon the duration of the interruption.Interruptions
¢
100 ms were detected by the cell as shown by elimination of field-induced enhance-ment of ODC. That two time constants (0.1 and 10 s) are involved in cellular EMF detection isconsistent with the temporal sensing process associated with bacterial chemotaxis. By analogy withbacterial temporal sensing, cells would continuously sample and average an EM field over intervalsof about 0.1 s (the ‘‘averaging’’ time), storing the averaged value in memory. The cell would comparethe stored value with the current average, and respond to the EM field only when field parametersremain constant over intervals of approximately 10 s (the ‘‘memory’’ time).
Bioelectromagnetics18:388–395, 1997.
1997 Wiley-Liss, Inc.
Key words: ornithine decarboxylase; cell culture; 60 Hz fields; ‘‘averaging’’ time; ‘‘memory’’time
INTRODUCTION
exposed these cells to a magnetic field whose frequencywas incoherently switched between 55 and 65 Hz atSpecific effects induced by exposure of cells toset intervals (coherence times). The resulting enhance-weak, extremely low frequency (ELF), electromagneticment of ODC activity depended upon the time durationfields (EMFs) continue to be reported but the mecha-of the switching interval. It was necessary to maintainnisms that underlie cellular detection of EM fields areeach frequency for approximately 10 s or longer tostill not understood. It is clearly important to determineobtain the same enhancement of ODC activity that wasthe specific molecular interactions involved in EMFinduced by exposure to a field of constant frequency.detection. Until such information is known, however,The possibility that this 10-s time constant re-aspects of the detection mechanism may be revealedby characterizing the parameters of EM fields that arecritical for initiation of a cellular response.
Contract Grant sponsor: National Institute of Environmental Health Sci-
To this end, we previously examined the question
ences (NIEHS), National Institutes of Health (NIH); Contract Grantnumber: 5 R01 ES0687-02.
of whether cellular response to an EM field requiresthat the field be coherent, i.e., that field parameters be
*Correspondence to: J. Michael Mullins, Department of Biology, The
predictable over time [Litovitz et al., 1991]. Magnetic
Catholic University of America, Cardinal Station, Washington, DC20064.
fields with frequencies of 55, 60, or 65 Hz induce adoubling of ornithine decarboxylase (ODC) activity in
Received for review 6 May 1996; final revision received 21 January1997
L929 cells. To assess the role of field coherence, we
1997 Wiley-Liss, Inc.
827d / 8507$$827d 06-04-97 12:12:45 bema W: BEM
 
Role of Temporal Sensing 389
flected a fundamental property of the cell’s ability to
MATERIALS AND METHODS
detect, and respond to, EMFs was intriguing. Nonethe-
Cell Culture, ODC Assay, and Statistical Analysis
less, some basic questions were unanswered. (1) WasMethods for the maintenance of L929 cell cul-this fundamental 10-s time constant related only totures, and for the assay of ODC specific activity wereshifts in field frequency or, as seemed likely to us,done following the procedures described by Litovitz etwould changes in other field parameters (e.g., ampli-al. [1994]. To provide blinded experimental conditions,tude, wave form) produce similar results? (2) In thesamples were coded by one individual so that neitheroriginal work, the time duration of each frequency in-the exposure condition to which each corresponded,terval was slightly randomized by adding to the basicnor the exposure system used to produce it, wereinterval a random period ranging from 0 to 50 ms. Thisknown to the person doing the ODC assays. After ODCrandomization produced phase incoherence, and wasactivities were calculated and recorded, this code wasthe rationale for our use of the term ‘‘coherence time’’‘‘broken,’’ and the results of the experiment were ana-to describe the switching intervals. The question re-lyzed. As in our previous work, the results of eachmained, however, whether the significant factor wasexposure were expressed as an ODC activity ratio, ob-truly the coherence.tained by dividing the activity of an EMF-exposed sam-Could it be that simply the constancy of fieldple by that of its matched, sham-exposed control. Be-parameters (e.g., frequency or amplitude) determinescause the ODC activities of control L929 cultures dis-whether there is a field-induced bioresponse? The work play day-to-day variation, use of the ODC activity ratioreported here addresses this question. The
amplitude
allows normalization of experimental results from oneof the applied magnetic field was varied at regularday to the next.intervals throughout EMF exposure, and the resultantWe previously demonstrated [Litovitz et al.,enhancement of ODC activity was assessed. For these1993, 1994] the consistency and effectiveness of theexperiments, field coherence was maintained by elimi-ODC activity ratio for determining the cellular re-nating the insertion of random time intervals uponsponse to applied EM fields. However, some scientistsswitching. Results of this work allow us to determinehave expressed reservations about the use of ratioswhether it is the coherence or the simple constancy of to determine whether EMF-induced responses exist.the applied field parameters that determines a cellularAccordingly, we have also included in this report theresponse.results of a statistical analysis done using a paired, two-In considering possible mechanisms for EM fieldtailed
test to determine the statistical significance of detection that underlie these data, we drew ideas fromeach experimental condition. Tables provided for eachthe well-characterized body of information on bacterialexperimental condition list the mean ODC activities,chemotaxis [Koshland, 1981; Segall, et al., 1982; Mori-
{
standard deviation, for both sham and exposed sam-moto and Koshland, 1991]. Motile bacteria use a rudi-ples. The paired
P
value for each condition was deter-mentary system of memory to sense average values formined with INSTAT, a statistics program distributedconcentrations of extracellular attractants and repel-by GraphPad Software, Inc. (San Diego, CA).lents, to compare a ‘‘remembered’’ average value witha current one, and to direct their swimming based upon
Exposure System
measured variation in the average concentration overtime. Functioning of this memory depends upon spe- All exposures were carried out using a Helm-holtz-coil type, ELF exposure system. Sham exposurescific interactions between two biochemical events, onethat occurs over a few tenths of a second (the averaging were concurrently carried out in such a system withthe field turned off. The Helmholtz coils, which are E-time) and one that occurs over intervals of a few sec-onds (the memory time). Given that our earlier work field shielded and have an average radius of 4.125inches, were enclosed in a 10
1
10
1
10 inch mu-metal[Litovitz et al., 1991] demonstrated a time constant onthe order of 10 s for the cellular-ODC response to an box constructed of mil-annealed Amumetal (AmunealManufacturing Corp., Philadelphia, PA). Measure-EM field, we wondered whether a memory functionmight also be involved in EM field detection and re- ments showed that the magnetic field shielding effi-ciency of this mu-metal box is approximately a factorsponse. To explore this possibility, a series of experi-ments was initiated to look for a second, shorter time of 5 in the ELF range of interest. Each Helmholtz coilunit was housed within a water jacketed, cell cultureconstant comparable to that of the averaging time of the bacterial system. The results of these experiments incubator maintained at 37
C.Each exposure condition was carried out usingare discussed in the context of a mechanism that wouldgovern cellular response to EMFs based on the con- three, stacked, 75 cm
2
tissue culture flasks that werecentered between the paired coils and were positionedstancy, over time, of EM field parameters.
827d / 8507$$827d 06-04-97 12:12:45 bema W: BEM
 
390 Litovitz et al.
so that the magnetic field was parallel to their growthsurfaces. The magnetic field distribution within the re-gion occupied by the cells varied by less than
{
15%from the nominal value at the center of the Helmholtzcoil.The Helmholtz coils were operated with a pulsemodulated 60 Hz sinusoidal current. The exposure sig-nal was produced using a function generator (TENMAmodel 72-380, MCM Electronics, Centerville, OH),either of two custom made pulse modulators, and a 35watt audio amplifier (Realistic model MPA-45, TandyCorporation, Fort Worth, TX). The Helmholtz coilsand a 3
V
series resistor were connected to the 8
V
speaker output of the amplifier. The pulse modulatorswere used to introduce periodic changes in the signalamplitude. One pulse modulator allowed switching of 
Fig. 1. Response of L929 cultures to 60 Hz fields, which were
the signal amplitude between zero and a fixed level,
turned off and on at an interval of 0.1, 1, 5, 10, 20, or 50 s
on successive equal time intervals of 0.1, 1, 5, 10, 20,
throughout exposure. Results are expressed as the mean orni-thine decarboxylase (ODC) activity ratios
{
standard deviation.
and 50 s. The other modulator allowed momentary
Numbersofseparateexperimentsperformedforeachdatapoint
interruption of the signal on successive 1-s intervals.
are given in Table 1.
The interruption interval was selectable in the range 0to 500 ms. Additionally, these devices were used toexpose cells to fields for which the amplitude was regu-larly switched between 5 and 15
m
T, providing a mean sponse to EMFs, we conducted a series of exposuresin which the amplitude of the applied 60 Hz field wasamplitude of 10
m
T during the course of exposure.The on-off transitions used in our experiments switched between 0 and 10
m
T. This was done at inter-vals,
t
c
, of 0.1, 1, 5, 10, 20 or 50 s throughout the 4-hresulted in the production of switching transients bythe inductive elements, chiefly the Helmholtz coils and exposure period. The field thus alternated between be-ing completely off and then on at an amplitude knownpower amplifier, in the exposure system. Because theinput signal to the amplifier was not synchronized for to induce enhanced ODC activity. In contrast to ourprevious experiments with frequency switching, therezero cross-switching, the transients were of variablepeak amplitude (less than 25% of the peak-to-peak was no randomization of the intervals by inclusion of additional, random time periods.signal amplitude). Their duration was less than 10 ms.For some exposures input signals were switched at zero The results of these experiments are shown inFigure 1. Cells exposed to fields that were turned off crossing points using an Arbitrary Waveform Genera-tor (Hewlett Packard model 33120A). In this case the and on at
t
c
intervals ranging from 5 to 50 s displayedstatistically significant enhancements in ODC activity.switching transients were of constant peak amplitude,approximately equal to 25% of the peak-to-peak signal Intervals of 10, 20, or 50 s produced comparable en-hancements, with ODC activity ratios of about 1.7.amplitude. The time duration of these transients wasalso of the order of 10 ms. ODC activities of cultures exposed to elds switchedat a 5-s
t
c
interval were also elevated, but to a lesserTo study the bioeffects of transient-free variationsin signal amplitude, ODC activity was examined for extent (ODC ratio
Å
1.4) than those obtained with thelonger
t
c
intervals. In contrast, a
t
c
interval of 0.1 orcells exposed to a 60 Hz signal whose amplitude wasvaried periodically by amplitude modulating the 60 Hz 1 s produced no statistically significant enhancementof ODC activity. A statistical analysis of the ODCsinusoid with a 1 Hz sinusoid. The modulation depthwas adjusted so that amplitude of the 60 Hz signal activity (see ‘‘Materials and Methods’’) obtained fromthese exposures is presented in Table 1. It can be seenvaried sinusoidally at 1 Hz between 0 and 10
m
T.from the table that only when
t
c
was 1 s or less wereODC activities statistically equivalent to those obtained
RESULTS
with sham exposure conditions. For longer
t
c
’s, p valueswere less than .05 indicating that the enhancement of 
 Amplitude Switching and the Role of Incoherence
ODC activity shown in Table 1 is statistically signifi-cant.To determine whether variations in field parame-ters other than frequency would affect the cellular re- These results are essentially the same as those
827d / 8507$$827d 06-04-97 12:12:45 bema W: BEM

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