Energy, Fitness, and Information-Augmented Electromagnetic fields in drosophila

Energy, Fitness, and Information-Augmented Electromagnetic fields in drosophila

(Parte 1 de 3)

Energy, Fitness, and Information-Augmented Electromagnetic Fields in Drosophila melanogaster

MICHAELJ. KOHANEDitron LLC, P.O. Box 70 Excelsior, MN

WILLIAMA. TILLERDepartment of Materials Science and Engineering, Stanford University Stanford, CA

Abstract—Exposure of developi ng larvae to a few specific electrom agnetic fields ( EMFs) and informatio n-augme nted EMFs modified (a ) t he expression of larval developm ent time, a genetica lly based trait relevant to developm ent and whole organism fitness, and (b ) a measure of energy metabolism, the [adenine triphosp hate/ adenine diphosp hate] ([ATP]/ [ADP] ) ratio in isofemale strains of Drosophi la melanoga ster . The study represents a compilat ion of approxi mately 10,0 larvae and 7,0 adults counted. The specific EM frequenc ies used in this study, 5.0, 7.3, 8.0, and 9.3 MHz at output power levels in the approxi mately 1 microwatt range, were produced by two small electro nic devices of physical ly identica l nature, but one was intentio nally imprinte d with specific informati on. Exposure periods varied from 4 hours to one life cycle. Larval developm ent time was significant ly shorten ed ( approxi mately 10%) and the [ATP]/

[ADP] ratio significant ly increased (approx imately 30%) in a Faraday cage without the EMFs compared to a Faraday cage with the specific EMFs. The Faraday cage represents a shielded environ ment that facilita tes exposure to both fewer and specific electrom agnetic frequen cies. Larval developm ent time results for developm ent in the laborato ry environ ment, which represents exposure to backgro und EMFs of various frequenc ies, were intermedi ate. The informatio n-augme nted EMFs also gave intermedi ate results. Overall, there were no signific ant effects observed for t he other measured fitness compon ents— third instar larval weight, adult survival , and surviving adult weight. We discuss a thermody namic model to account for our results and general bioelec tromagn etic effects and attribu te the change in developm ent time to EMFs modifying electro n transpo rt chain activity and the [ATP]/

[ADP] ratio via t he influence of the EMF/ magnetic vector potenti al upon electron availabi lity and nicotina mide adenine dinucle otide levels.

Keywords:fitness experime nts— [ATP]/

[ADP] ratio— Faraday cages— informati on-augm ented and normal EMFs, theoret ical models

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1. Introduction Bioelec tromagn etic studies have advanc ed conside rably in t he past decade (Goodma n, Greenbau m, and Marron, 1995 ). For exampl e, studies of nont hermal effects on cells of t he immune system from exposure to electro magnetic fields (EMFs ) in t he extremel y low frequenc y range (<300 Hz ) indicate t hat stimulat ory, inhibit ory, and no field exposu re effects exist even for identic al field paramete rs. The results depend upon the degree of cellular activat ion and t he physical and biochem ical boundar y condit ions experien ced (Eichwal d and Walleczek , 1996 ). Furt hermore, low frequenc y EM Fs influence specific RNA transcri pts in human cells and transcri ption in Drosophi la melanoga ster cells(Goodma n, Wei, and Henderso n, 1989; Goodman et al.,1992 ), and Ho et al.(1992 ) have shown that weak static magnetic fields cause abnormal ities in first instar larvae of D. melanoga ster .A number of models have been propose d to account for EMF effects on biological systems. Ho et al.(1992 ) suggested t hat t he weak static magneti c fields t hey studied must affect some coo perative process involved in pattern determination during critical stages of early Drosophila developm ent. On t he basis oft he immune cell experime nts, Eichwald and Walleczek (1996 ) suggested t hat external EMFs interact wit h cellular systems at the level of intrace llular signal transdu ction pathwa ys, specifica lly, Ca-sign aling processes. Nossol, Buse, and Silny (1993 ) reported magneti c field inf luences on t he invitro redox activit y of cytoch rome oxidase activit y. Additio nally, weak EM Fs stimulat e adenine triphos phate (ATP ) synthes is and alter Na,K-ATPase activity (Blank, Soo, and Papstein , 1995; Lei and Berg, 1998 ). Thus, magneti c fields may inf luence cellular energy metabol ism, and Menend ez (1996 ) has suggested t hat an electrom agnetic coupling process may explain t he proton transloc ation mechanis m in cellular energy transfer. A signific ant associat ion has been observed between stress, larval development time and aspects of energy metabol ism, t he cofactor nicotin amide adenine dinucle otide (NAD ), and the [ATP]/

[ADP] ratio in D. melanoga ster (Kohane , 1988, 1994 ). In t he present paper , we use t he t heoretica l and experimental approac h presented in these earlier papers and expand it to incorpo rate t he intenti on-im printed electro nic device (IIED ) techniq ues of Dibble and Tiller (1999 ) and investiga te t he hypot hesis t hat both normal and informati onaugment ed EM Fs may inf luence fitness and energy metabol ism. We study fitness and the [ATP]/

[ADP] ratio under nonstressf ul nutrien t conditions , in t he presence and absence of small electron ic devices t hat produce EM Fs of frequenc ies much higher t han t hose used in previou s studies. We assess t hese frequenc y effects using exposure periods from 4 hours to one life cycle in order to detect EMF effects that may not be observed at more conservative levels of EMF frequenc y and exposu re period. Our results indicat e t hat larval developm ent time is signific antly shorten ed and t he [ATP]/

[ADP] ratio signific antly increased in a Faraday cage wit hout t he devices com pared to a Faraday cage with t he devices. The Faraday cage

represents a shielded environ ment with respect to electrom agnetic (EM ) radiation and facilit ates exposure to fewer frequenc ies in t he absence of devices and specific EM frequenc ies in the presence of devices. In additio n, t he IIEDs gave signific antly better results on larval develop ment time t han did the unim printed control devices. Overall, t here are no signific ant effects for the ot her fitness com ponent s assessed, third instar larval weight, adult survival , and surviving adult weight. Thus, a reduction in exposure to EMFs increases one com ponent of fitness, suggestin g t hat EMFs may act as a biologic al stress (Goodma n, Greenbau m, and Marron, 1995; Smit h, 1996 ). Althou gh we acknowl edge t he fact t hat EM Fs may modify t he larval environ ment ( e.g., t he food ), we attribu te the effects of the specific EMFs in t his study to t he modified [ATP]/

[ADP] ratio as a consequ ence of altered NAD levels, electron availab ility, and electro n transport chain activit y. Finally, we discuss a t hermodyn amic model t hat may rationalize our observed fitness and energy changes and general bioelec tromagn etic effects.

2. Experimental Methods (a ) Strains We studied larvae obtained from two isofemale strains, Strains 1 and 2 of Kohane (1994 ). Nonstressfu l food was used for strain culture and experime nts, and t he food com positio n was as follows: 36 g agar, 108 g sugar, 72 g dry yeast, 10 ml propio nic acid, and 24 ml nipage n in 2,0 ml water (Kohane , 1987 ). Separat e constan t temperat ure rooms (18º C, 5% relative humidi ty ) were used for (a ) device storage, (b ) strain culture and unexpo sed adult culture, and (c ) experime nts.

(b ) Faraday Cages A standard Faraday cage consisted of a copper mesh screen enclosing a certain spatial volume. It is electric ally grounde d so the EM waves of waveleng th larger t han t he mesh size, which im pinge on t he screen, will leak off to ground and only minimal ly penetrat e to t he interio r space. Thus, the interior space has a greatly reduced EM integra ted power density in the wavelengt h range largert han the copper mesh spacing. The one layer of co pper mesh cages (dimensions: 40 40 30 cm ) used here can be expected to reduce t he EM field strength by a factor of approxi mately

(c) Electronic Devices Our experime nts used two electro nic devices in order to assess exposure to multipl e frequenc ies and a single frequenc y as follows: (d1 ), a triple oscillat or device produci ng frequenc ies of 5.0, 8.0, and 9.3 M Hz; and (d2 ), a single oscillator device, produci ng a frequenc y of 7.3 MHz. Additio nally, we studied two categor ies of EMFs produce d by these devices. The first category

Electro magnetic Fields in D. melanoga ster

Tiller

involved devices (d1, o ) and (d2, o ), which had not been exposed to human information al inf luences. The second category involved devices (d1, j ) and (d2, j ), which had been exposed to human informati onal inf luences (see below ). Thus, (d1, o ) and (d1, j ) and (d2, o ) and (d2, j ) constitu ted physica lly identica l pairs of devices t hat differed only in t he fact t hat one of each pair , (d1, j ) and (d2, j ), respective ly, had been exposed to t he human informatio nal inf luence. The devices were individ ually wrapped in aluminu m foil and stored in separate Faraday cages and were fabricat ed to be identic al to t hose produce d by Clarus Corporat ion (La Costa, CA ). The triple oscillat or device was powered by line voltage to 9V DC, and t he single oscillat or device was previou sly charged for 24 h by line voltage to 9 V DC and used wit h battery power.

(d ) Intentions The actual im printing procedure was as follows: (a ) The device was placed along wit h its current transforme r on a table around which t he imprint ers sit; (b ) Four peo ple (two men plus two women ) who were all accom plished meditators, coherent , inner-sel f managed and readily capable of entering an ordered mode of heart functio n (Tiller, 1997 ) and sustaini ng it for an extende d period of time, sat around the table ready to enter a deep meditat ive state; (c ) A signal was then given to enter such an internal state, to cleanse the environ - ment and create a sacred space for t he intenti on, then, a signal was given by one of t he four to put attenti on on the tableto p objects and begin a mental cleansing process to erase any prior imprint s from t he device; (d ) After 3 or 4 minutes, anothe r signal was given to begin focusing on t he specific prearranged intenti on statemen t for about 10– 15 minutes; (e ) Next, a final signal was given to shift focus to a closing intenti on designed to seal off t he imprint into t he device and minimiz e t he leakage of t he essentia l energy/ informati on from t he devices. This com pleted t he process, so t he four peo ple withdrew from the meditat ive state and returned to their normal state of conscio usness. The specific intenti on was “ to activat e t he indwell ing conscio usness of t he device (d, j ) so as to increase t he concen tration of NAD plus t he activit y of t he availabl e enzymes, dehydro genases and ATP syntha se in t he mitocho ndria so that product ion of ATP is signific antly increased relative to that produce d in the unimpr inted device (d, o ).” This chemical transforma tion process in t he cells of t he fruit f ly larvae was expected to signific antly inf luence their fitness, which would manifest itself via a reduced larval develo pment time for t hese larvae because they have a larger pool of ATP to work with (Kohane , 1994

(e ) Fitness We conduc ted four similar experime nts over a 6-mont h period and assessed EMF effects on t he fitness of t he strains using the above devices and different exposure periods. The experime nts are summariz ed in Tables 1 and 2. We measured fitness at 18º C using t he procedu res given in Kohane (1988; 1994

Electro magnetic Fields in D. melanoga ster

Experime nts were conduc ted at 18º C since developm ent time differenc es have been detected at this temperat ure for t he strains studied here (Kohane , 1994 ). The treatmen ts investiga ted in t he experime nts were as follows: (a ) (C )— culture in t he random EMF environ ment of the laborato ry; (b ) (F )— culture in t he relativel y reduced EMF environ ment of t he Faraday cage wit hout a device; (c ) (d1, o ) and (d2, o )— culture in t he relativel y reduced EMF environ ment oft he Faraday cage in t he presence of a device t hat had not been associat ed with human informati onal inf luences; and (d ) (d1, j ) and (d2, j )— culture in t he relatively reduced EMF environ ment of the Faraday cage in t he presence of a device that had been associat ed wit h human informati onal inf luences. A single replicate involved 30 larvae (0– 4 h old ) transferre d to a single vial contain ing nonstressf ul food (see above ). For each experime nt, all vials were establishe d within a 3-hour period. Vials were transferre d to Faraday cages and t he cages were placed, at the same time, immediat ely next to each ot her on t he same bench in a constan t temperat ure room. Treatment (C ) involved vials concurrently transferre d to a tray, which was placed on t he lid of treatmen t (F ). We used vial cultures as a thermoc ouple and could not detect temperat ure variation between treatmen ts in each experime nt. Exposure of vials to devices in Faraday cages was achieved as follows: The

TABLE 1Summary of Fitness Experiments Treatment and replicat e number ExposureExperime nt Strain aDevicebCFcd, jdd, odperiodeDate12d1 16 15 15 16 4 hours February 1997 2 1 d2 16 15 15 16 4 hours February 1997 3 1 d1 15 14 15 15 4 days July 19974f,g1d1 15 15 16 16 Life cycle plus July 1997 4 daysNote: Fitness componen ts assessed were larval development time, adult survival, and surviving adult weight. a Strain 1 and strain 2 refer to two isofemale strains. b (d1 ) refers to a triple oscillator device producing frequencies of 5.0, 8.0, and 9.3 MHz; and (d2 ) to a single oscillator device, producing a frequency of 7.3 MHz. The output power of the devices at the exposure distances is expected to be less than 1 µ W. Categories of EMFs produced by these devices were as follows: (a ) (d, o ), devices which had not been exposed to human informational influences; (b ) (d, j

), devices which had been exposed to human informationa l influences— an intention concerned with significant ly increasing the [ATP]/ [ADP] ratio and decreasing larval development time. c (F ) refers to culture in a Faraday cage without a device, and (C ) refers to culture in the laboratory environmen t. d (d, o ) and (d, j ) refer to culture in a Faraday cage with a single device. e Experiments were conducted at 18º C. Experimental vial cultures involved 30 larvae (0– 4 h old ) transferred to a single vial containing nonstressful food. f For Experiments 1– 3, larvae were derived from unexposed adults, and for Experiment 4, larvae were derived from exposed adults. g The [ATP]/

[ADP] ratio and third instar larval weight were assessed in Experiment 4 (see Materials and Methods

Tiller

respective device was placed in t he center of a Faraday cage and vials/ bottles were transferre d to cages and placed around t he perimete r. The device was t hen removed at t he end of a specific time period and larval develo pment proceeded until adults eclosed. The device was approxi mately 15 cm from t he vials/ bottles on average, where t he output power of the devices in their specific frequenc y ranges is expected to be less t han 1 microwatt . Vials were monitored daily and surviving adults were collect ed and weighed. Surviving adult weight was calculat ed for each vial as t he weight of t he number of flies survivin g divided by t he number of f lies surviving and is given in mg. Larval develo pment time is given as T, the time taken for half of t he surviving adults to emerge.

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