Research of Electrical Current Against Viruses

Notes on the following papers test methods:
Human CD4 immune blood cells were exposed to various amounts of electric current for various amounts of time and then mixed with HIV-1 virus. The resultant decrease in CD4 infection by the virus (resulting in cell enlargement [syncytium] and reverse transcriptase production) is listed in the two tables under the column headings indicating how much electric current in microamperes (ľA= millionths of an ampere) was applied to the virus for how many minutes. 100ľA applied for 3 minutes to the virus produced only approximately 2% of the number of enlarged cells (syncytium) as was produced after 6 minutes without electrical treatment of the virus. 100ľA is equal to 7.7ľA per square millimeter (mm²) of the surface of the electrodes delivering the electric current to the virus. Explanation of tests: Human CD4 immune cells, when exposed to HIV-1 virus, normally become infected with the virus and then enlarged as the virus reproduces itself (with the coerced help of the blood cell) within the CD4 cell. So anything that can reduce the viral infectivity (the ability of the virus to infect a cell) can be tested in the laboratory to see if less enlarged cells (syncytium) are produced which will show the degree of success of the anti-viral. Reverse transcriptase is an enzyme used by retroviruses to form a complementary DNA sequence (cDNA) from their RNA. The resulting DNA is then inserted into the chromosome of the host cell. A test showing how much is being produced is a way to quantify approximately how much viral reproduction has happened as a result of viral infection of the CD4 cells. So a test result showing reduced reverse transcriptase activity is one that shows how successful an anti-viral is at reducing the infectivity of a virus.

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BIOCOMPATIBLE ELECTRIC CURRENT ATTENUATES HIV-1 INFECTIVITY

William D. Lyman, Irwin R. Merkatz
William C. Hatch and Steven C. Kaali
Departments of Pathology, and Obstetrics & Gyneclogy
Albert Einstein College of Medicine,
1300 Morris Park Ave. Bronx, N.Y. 10461

Summary
In this report, we present the results of double-blinded studies on the use of direct electric current to alter the infectivity of HIV-1 for susceptible cells in vitro. These experimental currents were equal to 3.85 and 7.7ľA/mm² current densities respectively. The reduction of infectivity was dependent upon the total electric charge (ľA x min) passing through the chamber to which the virus was exposed. Viral infectivity was determined by two independent measures: a syncytium-formation assay which can be used to quantify the production of infectious particles; and a reverse transcriptase assay which is an index of viral protein production.

Results

Syncytium-formation assay:
Using this index of HIV-1 infectivity, it was determined that exposing virus to direct electric current suppressed its capacity to induce the formation of syncytia. Figure 1 shows a representative experiment and Table 2 shows the Croup data for 3 separate experiments. As can be noted in Figure 1, a statistically significant (p<0.001) reduction in syncytium number was absented and this reduction was dependent upon the current applied to the viral isolate. At three different viral dilutions, there were analogous results in that a total charge of 200ľA x min (25ľA for 8 minutes) reduced the number of syncytia from 50% to 65% while a charge of 3O0ľA x min (50ľA for 6 minutes, 75ľA for 4 minutes or 100ľA for 3 minutes) resulted in 90% reduction.

Reverse transcriptase assay:
The direct electric currents to which HIV-1 was exposed also reduced reverse transcriptase activity. Five separate experiments were conducted and a representative experiment is shown in Figure 2 and the data are included in Table 3. As can be seen in Figure 2, there was a significant decrease in the amount of reverse transcriptase activity after exposure of the virus to either 50ľA for 3 or 6 minutes. An equivalent reduction in reverse transcriptase activity was also noted with exposure to 100ľA for 3 minutes and almost ablation of reverse transcriptase activity was seen with exposure of the viral isolate to 100ľA for 6 minutes. The group data (Table 3) show that after exposure to 50ľA for 6 minutes, there was a 44% reduction in activity and treatment of virus with 100ľA for 6 minutes resulted in a 94% reduction. An analysis of variance indicates that the decrease in reverse transcriptase activity was statistically significant (p<0.0001).

DISCUSSION
The results reported here demonstrate that HIV-1 treated with direct electric currents from 50 to 100ľA has a significantly reduced infectivity for susceptible cells in vitro. This reduction of infectivity correlates with the total electric charge passing through the chamber. The therapeutic potential of electric current may reside in its ability to lower the viral titer to subclinical significance or in its incorporation into a strategy analogous to that of other therapies in which repeated cycles of treatment eventually achieve remission or cure. It may be also feasible to treat AIDS patients with direct electric current using either extracorporeal [out of body] systems or self contained indwelling electrodes.

Figure l

Syncytium Formation at 1:160 dilution

exposure (ľA/minute)    0/6   25/8   50/6   75/4   100/3   
# of syncytium formed   128    45      9      7      2    

exposure (ľA/minute)  0/3   0/6   50/3   50/6   100/3   100/6   
reverse transcriptase 300   205    108    105    100      12   count per million x 10-3 

Notes on the following patents test methods:
The patent's 1st laboratory test used electrically treated (at 100ľA/3min which is 7.7ľA per square millimeter for 3 minutes) HIV-1 virus mixed with human T cells (immune cells which the HIV likes to infect) and showed 99% less infected & enlarged T cells (syncytia) by viruses after 4 days compared to T cells mixed with HIV-1 not electrically treated. The patents 2nd lab test used electrically treated (l00ľA/6min) HIV-1 mixed with blood T cells and showed 72% less virus associated enzymes (reverse transcriptase produced as a result of viral reproduction) after 4 days, compared to T cells mixed with HIV-1 not treated. The patent stated; "it is possible that certain virus may be attenuated (or attenuated at a faster rate) if they are exposed to a greater electric current magnitude of the order of 500 microamperes for shorter time periods." 500 microamperes (500ľA) applied to the experiments electrodes with a surface area of 13.3mm² yields a current of 38.5ľA per square millimeter (38.5ľA/mm²). In approximation of this idea, my Microbe Electrifier was estimated to produce 17 to 34ľA/mm² (for 4 - 8 mm² cross sectional arterial area). This high current is necessary due to the short (but repeated) treatment durations of blood when using an electrode on each wrist. Due to the short durations of exposure to the electricity, as the blood passes through the main arteries, it is necessary to do treatments of up to 1 - 2 hours.

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U.S. PATENT 5,139,684
By Steven Kaali and Peter Schwolsky
Filed 11-16-1990, approved 8-18-1992

Electrically conductive methods and systems for treatment of blood and other body fluids and/or synthetic fluids with electric forces

Claimed:
"...applying... no biologically damaging electric potentials... to the electrically conductive electrode segments whereby electric field forces are produced... that induce biologically compatible current flow through the blood... to attenuate bacteria, virus, parasites, fungus contained in the blood... to render the bacteria, virus, parasites, fungus ineffective while not impairing the biological usefulness of the fluids"

Summary:
"...to attenuate such contaminants to the extent that bacteria, virus, or fungus, or parasites contained in the blood... are rendered ineffective to infect or affect healthy cells."

Best Mode of Practicing Invention:
"For example, treatment of AIDS virus in media at 100 microamperes for 3 minutes has been observed to substantially attenuate (render ineffective) the AIDS virus."

Experimental Results:
"In conclusion, these experiments... indicate at a statistically significant level that direct electric current... can attenuate the ability of HIV-1 to infect normally healthy cells which are susceptible to the HIV-1 AIDS virus." "Both of these assays (syncytia & reverse transcriptase) are widely used as reproducible measures of viral infection and can be used to determine if alternations in viral infectivity as a product of this electrical treatment can be detected." "...it is believed obvious that other modifications and variations of the invention will be suggested to those skilled in the art in the light of the above teachings." "...it is possible that certain virus may be attenuated (or attenuated at a faster rate) if they are exposed to a greater electric current magnitude of the order of 500 microamperes for shorter time periods."