BIOELECTROMAGNETIC MEDICINE
VOLUME EIGHT
JULY 2003
I. CLINICAL STUDIES IN BIOELECTROMAGNETIC MEDICINE
VOLUME EIGHT
JULY 2003
I. CLINICAL STUDIES IN BIOELECTROMAGNETIC MEDICINE
EFFECT OF ELECTRIC FIELDS ON THE BLOOD OF RAT-SPRAGUE DAWLEY
Manisha Mukewar and V.V. Baile*
ABSTRACT :
INTRODUCTION :
MATERIAL AND METHODS
The Exposure Facility and Mouse Cages :
The rats were housed in standard polypropylene rat breeding cages with a stainless steel grill (De Bruyn and De Jager, 1994; Oroza et al., 1987). These cages with animals were then kept in Faraday's cages for experimentation. The experimental and control groups of animals were kept apart. The control cages were kept at sufficient distance from experimental cages, so that there should not be any interference of exposure field. All other conditions of the control group were same as that of the experimental cages except the exposure field.
The husk was scattered in the plastic cages to form a bedding for rats. This material is very good absorbent. The food pellets made by Hindustan Lever Limited (HLL), India were given to the animals. It consists of all the nutrient values necessary for the healthy growth of rat. The rats had free access to water from water bottles.
The husk bedding was changed three times a week when the cages were washed with ordinary commercial dish washing powder. The cages were rinsed several times with running tap water in order to clean the residue of washing powder. Then the cages were kept for complete drying.
The Experimental Animal :
The Experimental Design :
Local university ethics committee has approved the protocol of said work.
OBSERVATIONS AND DISCUSSION :
More than 99% of the cells in the blood are red blood cells or erythrocytes. Life span of red cells circulating in the blood is about 60 to 70 days in rats (Feldman et al., 1997). Total R.B.C. count underwent significant reduction in females in 1.5 Kv/m exposure (P<.01) and in 10 Kv/m exposure (P<,0.05) on day 30. Along with this there was significant reduction in haemoglobin content specially in females upto 120 days. Reduction in R.B.C. count and haemoglobin content clearly indicated setting of anaemic conditions. Males also exhibited decreased R.B.C. count and reduction in haemoglobin content but the values were mostly non significant. Haemoglobin accounts for 92-98% of total blood iron and in human beings it corresponds to about 50 mg of inorganic iron per 100 ml of blood (Chatterjee, 1980). When iron is not available in sufficient quantity, red cells can be formed, but they lack normal content of iron containing pigment and anaemia characterized by a low mean corpuscular haemoglobin concentration (MCHC) results (Bell et al., 1972).
In the present study, MCHC values in both males and females exposed to 1.5 Kv/m electric field and in the males of 10 Kv/m exposed fields showed significant reduction on 30 days exposure, indicating anaemic conditions setting in. Reduction in erythrocyte count, hemoglobin and haematoerit values were also noted in the males of Swiss Mice (Bonhomme Faivre et al., 1998) when exposed to electromagnetic field. Both marked reduction in colony forming ability of erythroid and myeloid processor cells in mice exposed to electric field for 9 days was also reported by Huang and Mold (1980). The resulting anaemia was possibly due to foliate and vit. B12 deficiency or reduction in iron metabolism and was of macrocystic type.
Hematocrit or packed cell volume (PCV) vary tremendously depending upon whether the individual has anaemia or upon his bodily activity (Hurkat and Mathur, 1976). In the females of Sprague dawley, upto 90 days significant decrease (P<0.05) in PCV both in 1.5 Kv/m and 10 Kv/m exposure was found but afterwards there was a significant increase in values on 120 days (P<0.05). Decrease upto 90 days strengthens the fact that anaemia would have set in but sudden increase thereafter could be attributed to dislocation of body fluids as reported by Villa et al., (1991), Warnke, (1980) and Sieron et al., (1989). Values for MCHC, MCH and MCV were also slightly reduced. MCV gives the insight of type of anaemia. Rat MCV can be affected by age, stock, sex and health status (Godwin et al., 1964; Hulse, 1964; Vondruska and Greco, 1973). In Sprague Dawley as MCV values did not show significant variation, the type of anaemia characterized is normocystic. Blood platelets initially upto 30 days were significantly reduced in males (P<0.01) and in females but afterwards there was not much variation in both the sexes.
Leukemia is characterized by uncontrolled cell proliferation (Svedenstal and Jahanson, 1998). Leukemia risk associated with electric field and electric field carrying greater risk than magnetic field was reported by Miller et al., (1996). In the present study, increase in the number of leukocytes is noted. Though the present studies are insufficient to demonstrate leukemia, it can be related with other studies reported, such as increase in the risk of leukemia among Ontario electric utility workers (Kheifets and Kelsey, 1996; Kheifets
et al., 1997; Villeneuve et al., 1989) and leukemia in French utility workers (Guenel et al., 1996) etc.
On the basis of reduction in total number of R.B.C. along with reduction in haemoglobin and almost normal values of MCV, the type of anaemia characterized for Sprague dawley upon exposure to electric fields was normocystic.
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Department of Zoology, Nagpur University, Nagpur-440 033, India.
Females and males of rat Sprague dawley, were subjected to electric fields of 1.5 Kv/m and 10 Kv/m strengths. Effect on various parameters of blood were studied and reported at the end of 30, 60, 90 and 120 days. Leukocytes upon exposure to 1.5 Kv/m electric field were initially decreased up to 30 days but afterwards there was significant increase in their total count from 90 to 120 days. In 10 Kv/m exposure, there was consistent increase indicating leukocytosis. Erythrocyte reduction and decrease in haemoglobin and haematocrit were more pronounced in females compared to the males indicating anaemic conditions; but values of mean corpuscular volume were almost the same as that of the control group. Thus the type of anaemia characterized here was normocystic.
The use of 'electricity' has brought tremendous technological advances in 20th century. Though the quality of life has improved, there is massive increase in the production and consumption of electric power which has resulted in increased environmental exposure to electromagnetic field both at work and in home (Cridland, 1997 a). It is clear that 60 Hz electric field (EF) is perceptible by rodents (Stern et al., 1983; Stern and Laties, 1985 and Sagan
et al., 1987), non human primates (Orr et al., 1995) and human (Deno and Zafenella, 1982; Kato et al., 1989; Cohen et al., 1992). Given the implications of stress hypothesis offered originally by Graves et al., (1977) and Marino
et al., (1980) and also a number of epidemiological studies suggesting increased risk of diseases (Valberg, 1996), a study was conducted in our laboratory subjecting rats - Sprague dawley to 1.5 Kv/m and 10 Kv/m electric fields. A number of haematological parameters were studied and reported upto 120 days. The similarlity of detection thresholds among rats, baboons and human support the contention that animal models are useful surrogates for humans in EF exposure studies (Orr et al., 1995). The actual detection threshold varies in different animals. Stern et al., (1983) reported that for 60 Hz electric field, the detection threshold for male rat was between 4 Kv/m and 10 Kv/m and for female rats it was between 3 Kv/m and 10 Kv/m (Stern and Laties, 1985). The exposure fields selected for the present study were 1.5 Kv/m and 10 Kv/m. It is also reported that 5% of the humans standing beneath a 60 Hz power line could detect 1 Kv/m, the median value for EF detection was 7 Kv/m (Deno and Zafanella, 1982). Prolific use of television sets, radio, power lines, air conditioning systems, heat from automobile exhaust and various pollutants causes the imbalance in air ionization in urban environment. Studies on blood of human volunteers in laboratory have shown that small positive air ions increase the amount of serotonin in blood while small negative air ions have opposite effects (Monaco and Ackar, 1963; Panati, 1980; Charry and Hawkinshire, 1981; Sulman, 1984; Smith and Best, 1989; Henshaw et al., 1996). Thus blood is an important liquid tissue which is responsible to carry all products of metabolism to various parts of body. Haematological alterations can lead to significant immunological effects. In the present study, therefore, emphasis is given on the study of various parameters of blood, because apparent epidemiological and experimental evidence available is controversial.
The Laboratory :
The experiments were conducted in the experimental animal unit of Department of Pharmaceutical Sciences, Nagpur University, Nagpur. Control and experimental rats (Sprague dawley) were housed in one room and thus exposed to the same environment, barring the exposure fields. The temperature of the room was maintained at 21-23o C, relative humidity noted was 50% to 70% and a light/dark schedule of 12/12 hours. In summer, coolers were installed in order to maintain the temperature. Laboratory facilities were set as prescribed by Farris and Griffith (1962).
In the present experiment, the cages were made of transparent PVC container with holes in all the directions for air circulation and were fixed in wooden frame. Two parallel metal plates were fixed at the bottom and top of the cages, having their back sides covered with fine plywood. This cage is called as Faraday's cage.
Healthy rats of species Sprague dawley were collected from the animal house of the Department of Pharmaceutical Sciences, Nagpur University Nagpur. The rats of approximately 60-90 days age and weight of 125-150 gms. were selected for experiments. Daily dairy was maintained to record the general health and mortality of the animals.
A number of males and females were kept separately for acclimatization for one week in the experimental room. After one week, the animals were used for experimentation. The cages for 1.5 Kv/m and 10 Kv/m and control cages of males and females were kept separately. The rats were kept for different exposure time such as 30 days, 60 days, 90 days and 120 days. At the end of each exposure time, blood samples for haematological studies were taken out from control as well as from experimental animals. Blood was withdrawn from medial venous plexus of the eye (Svedenstal and Johanson, 1998) and immediately processed for haematological investigations which were done on fully automated Coulter Cell Counter, England. The data is summarized in various tables. All statistics presented in this study are mean ± standard error. Student's 't' test was made use of for testing the significance of difference between the means of readings of experimental and control groups, using 5% level of significance.
Leukocytes or white blood cells upon exposure to 1.5 Kv/m electric field showed significant decrease in number both in males (P<0.001) and in females (P<0.05) initially upto 30 days, but afterwards there was significant increase in their number, as the duration of experiment increased from 90 to 120 days. Increase in total leukocyte count was however observed in 10 Kv/m exposure group consistently from 30 to 120 days both in males and females. Oroza et al., (1987), also reported increase in the number of leucocytes after 4 weeks exposure in male Wistar rats exposed to vertical electro magnetic field. A positive association between leukemia and combined exposure to electric and magnetic fields was also seen in non-human primates (Savitz et al., 1988). Leukocytosis was predominant in the present study, however, contradictory reports are available on the effects of electromagnetic fields on leukocytes as significant fall in leukocytes was reported on day 90 in Swiss Mice by Bonhomme - Faivre et al., (1998) and in mice exposed to 50 Hz electric field (Strampfer et al., 1979).
Bell, G.H., Davidson, J.N., Emslie-Smith, D. (1972) "Textbook of physiology and biochemistry". Publisher : The English Language Book Society and Churchill Livingstone.
Table 1
White Blood Corpuscles (W.B.C.) / Leukocytes - ul
------ ------ Male ------ ------ Female ------
Exposure Time Control 1.5 Kv/m 10 Kv/m Control 1.5 Kv/m 10 Kv/m
30 days 11980 7318(P0.001) 13536(n.s.) 6930 5480(P0.05) 8526(ns)
60 days 9060 8240(ns) 9300(ns) 7940 7880(ns) 8300(ns)
90days 9920 13140(P0.05) 12960(P0.05) 10180 14560(P0.05) 14480(P0.05)
120 days 9700 13800(P0.01) 13460(P0.05) 10180 15780(P0.05) 16000(P0.05)
All P are less than value in ( )
Table 2
Red Blood corpuscles (R.B.C.)/Erythrocytes-million/ul
------ ------ Male ------ ------ Female ------
Exposure Time Control 1.5 Kv/m 10 Kv/m Control 1.5 Kv/m 10 Kv/m
30 days 7.730 7.806(ns) 7.260(P0.01) 8.688 7.186(P0.01) 6.324(P0.05)
60 days 7.640 7.488(ns) 7.370(ns) 7.874 7.776(ns) 7.732(ns)
90 days 7.330 7.052(ns) 7.064(P0.05) 6.986 6.826(ns) 6.688(ns)
120 days 7.124 6.872(ns) 7.064(ns) 6.974 6.822(ns) 6.838(ns)
All P are less than value in ( )
Table 3
Haemoglobin (Hgb)-gm/dl.
------ ------ Male ------ ------ Female ------
Exposure Time Control 1.5 Kv/m 10 Kv/m Control 1.5 Kv/m 10 Kv/m
30 days 14.460 14.820(ns) 14.440(ns) 16.540 14.160(P0.05) 12.500(P0.05)
60 days 15.260 15.100(ns) 15.020(ns)> 16.020 15.340(P0.05) 15.420(ns)
90 days 13.940 13.710(ns) 13.640(ns) 14.960 13.660(P0.001) 12.820(P0.001)
120 days 16.400 15.680(ns) 15.400(P0.05) 14.960 14.780(ns) 14.800(ns)
All P are less than value in ( )
Table 4
Haematocrit(HCT)/packed cell Volunce(PCV)-%
------ ------ Male ------ ------ Female ------
Exposure Time Control 1.5 Kv/m 10 Kv/m Control 1.5 Kv/m 10 Kv/m
30 days 45.440 48.880(ns) 50.620 45.060(ns) 40.900(P0.05)
60 days 45.400 44.960(ns) 45.360(ns) 48.220 47.480(ns) 47.480(ns)
90 days 40.760 41.940(ns) 39.440(ns) 41.980 40.500(P0.05) 39.220(P0.05)
120 days 45.140 45.780(ns) 42.580(ns) 34.900 40.620(P0.05) 39.500(P0.05)
All P are less than value in ( )
Table 5
Mean Corpuscular Volume(MCV)-fl.
------ ------ Male ------ ------ Female ------
Exposure Time Control 1.5 Kv/m 10 Kv/m Control 1.5 Kv/m 10 Kv/m
30 days 57.540 62.680(P0.05) 62.680(ns) 61.820 61.600(ns) 64.920(ns)
60 days 60.340 58.680(ns) 59.040(ns) 62.220 61.820(ns) 60.520(P0.05)
90 days 52.700 53.340(ns) 54.700(ns) 57.520 55.700(P0.001) 56.980(ns)
120 days 55.180 53.760(ns) 52.300(ns) 54.860 54.380(ns) 53.920(ns)
All P are less than value in ( )
Table 6
Mean corpuscular Haemoglobin(MCH)-pg
------ ------ Male ------ ------ Female ------
Exposure Time Control 1.5 Kv/m 10 Kv/m Control 1.5 Kv/m 10 Kv/m
30 days 18.520 19.060(ns) 19.900(ns) 20.220 19.428(ns) 19.540(ns)
60 days 19.880 19.600(ns) 19.080(P0.05) 20.160 19.840(ns) 19.720(ns)
90 days 18.680 18.700(ns) 18.780(ns) 19.780 19.400(ns) 19.280(ns)
120 days 18.640 17.940(ns) 18.880(ns) 18.600 18.840(ns) 18.860ns
All P are less than value in ( )
Table 7
Mean corpuscular Haemoglobin concentration(MCHC)-gm/dl
------ ------ Male ------ ------ Female -------
Exposure Time Control 10 Kv/m 1.5 Kv/m 10 Kv/m
30 days 32.580 30.400(P0.001) 31.240(P0.01) 33.040 31.540(P0.01) 30.700(ns)
60 days 33.700 33.200(ns) 33.380(ns) 33.460 32.920(ns) 32.960(ns)
90 days 34.140 34.140(ns) 33.940(ns) 34.980 34.900(ns) 34.680(ns)
120 days 34.680 34.060(ns) 34.220(ns) 35.700 34.620(ns) 33.700(ns)
All P are less than value in ( )
Table 8
Platelet-lacs/ml
------ ------ Male ------ ------ Female -----
Exposure Time Control 1.5 Kv/m 10 Kv/m Control 1.5 Kv/m 10 Kv/m
30 days 8.344 6.706(P0.01) 6.154(P0.01) 8.504 7.310(ns) 7.646(ns)
60 days 7.720 7.566(ns) 7.538(ns) 7.568 7.748(ns) 7.228(ns)
90 days 7.692 7.680(ns) 7.524(ns) 7.410 7.212(ns) 7.418(ns)
120 days 7.662 7.6620(ns) 7.514(ns) 7.406 7.210(ns) 7.416(ns)
All P are less than value in ( )
ns = non-significant
A) The Energetic State (the Energetic Report)
The purpose of E.A.V. is to establish an Energetic Evaluation, a Functional Testing of organs and tissues through
the measure of Acupuncture and ElectroAcupuncture points in order to determine energetically unbalanced points.
We measure the conductance (capacity to let the stimulation current through) of an organ or a tissue in order to
discover energetically unbalanced points knowing that the energetic equilibrium of the human organism is altered,
among other things, by the negative ambiance influence exercised by some medications, poisons, insecticides,
viruses, bacteria, harmful electromagnetic fields and inflammations as well as certain aliments.
The body is the emitting and receiving focus of electromagnetic messages. Cells, as well as the entire organism,
constitute what is called in electronic an oscillatory circuit which is capable, if it is submitted to
electromagnetic waves, to reach resonance with one of these waves, that is the one that corresponds to the frequency
of the circuit. In fact, a very small stimulation current (of about 1 Volt and 10 MicroAmpere) is sent through the
acupuncture point. The result of the measurement can be observed on a gauge. The ideal measurement is 50. Depending
on the results observed on the gauge (graduated from 1 to 100).
1° QUANTITATIVES (from 0 to 100)
2° QUALITATIVES (with needle drops)
When we measure points, the needle does not stay stable at its apogee and start to fall (more or less rapidly)
towards zero. It generally signifies that there exist an Energetical Disequilibria at the level of the meridian
being measured. These measurements are to be preferred.
The ensemble of these measures (QUANTITATIVES + QUALITATIVES) permit to establish the ENERGETIC LEVEL of the
person.
B) Manifestation of the Accord of Frequencies
We can thereafter restore energetic imbalances with the addition of products and by the MANIFESTATION OF THE ACCORD
OF FREQUENCIES between the needs of the person and the product introduced in the circuit of measure.
That is what the Dr. Voll called ‘the Medicament Test.’ He observed that by depositing a product in the circuit and
if that was the appropriate product, it will correct the fall from the needle and will help the body to
re-equilibrate itself.
The most important point of that discovery is that a vibrating information, representing the product, is transported
or conveyed by the electric current following the acupuncture meridian to the organ in question.
In practice, it means:
When testing a product on a disturbed point (with fall of the needle), the following observations can be found after
placing the product to be tested in the circuit:
C) Control of Product Taking
Furthermore the EAV technique permits control very quickly if a given product, at the moment of the test, is IN
HARMONY with the needs of the person being tested. This allows continued control if a person keeps taking the
vitamin, dietary product, mineral element, etc.
In practice, it means that when testing a product on a stable point (without fall of the needle), the following
observations can be found after placing the product to be tested in the circuit: Voll, R: Twenty years of electroacupuncturein Germany: a progress report. Am.J. Acupunct 3:7-17,1975
EAV Links
Voll, R: The Phenomenon of medicine testing in electroacupuncture according to Voll. Am.J. Acupnct. 8:97-104,1980
The Mechanism of Acpuncture - Beyond Neurohumoral Theory, by Charles Shang, M.D. An excellent article that gives the reader a very
good theoretical description of the physiolgical basis of Acupuncture that is applicable to Bioelectromagnetic Medicine. The article may be found
here. The article has an excellent bibliography.