Investigation of the effect of a multi wave oscillator in the treatment of adult HIV/AIDS patients:

Authors: Campbell N R, Paspaliaris V, Ballard R

Enquiries: Prof. N. Campbell, P O Box 137, Parkville. Vic. 3052 Australia Email: noelc@smile.org.au

Abstract

In this study 26 patients infected with the HIV-1 virus were treated daily for two months with an electrotherapy device and their results compared with 27 non-treated patients matched for sex and age who were not participating in any anti-HIV treatment.

Over the 2 months of the trial, CD4 cell numbers reduced by 12% in the non-treated patients. The treated group showed a 5% increase in CD4 cell numbers. CD8 cell numbers dropped by 17% in the non-treated patients, and increased by 18% in the treated patients, over the same 2 months. In addition HIV viral loads increased by 109% in the untreated patients, and decreased by 6% in treated patients.

In many parts of the world where HIV infection is particularly rife, there are insufficient financial resources to pay for treatment with anti-viral drugs. This electrotherapy technique appears to be a low cost method for treating HIV infected patients who cannot afford conventional anti-viral treatment, and so are currently receiving no treatment at all.

Further investigation of this electrotherapy methodology in large-scale multi-centre trials is warranted.

Introduction

It is well known that human immunodeficiency virus (HIV) infection leads to depressed cellular immunity, which can result in serious opportunistic infections in acquired immune deficiency syndrome (AIDS) patients (Gottlieb et al. 1981; Gallo et al. 1984). At present, combination anti-retroviral drug regimens including protease inhibitors are used as a standard therapy for HIV-1 infection.

Access to antiretroviral drugs for HIV-infected patients in developing countries is a global public health priority. With the support of multilateral and bilateral programmes, non-governmental organisations, and national authorities, WHO has the ambitious objective to treat 3 million people with highly active antiretroviral therapy (HAART) by 2005 (WHO, UNAIDS 2003). WHO currently recommends first-line therapy with two nucleoside reverse transcriptase inhibitors (NRTIs) and one non-NRTI (NNRTI), a combination with good efficacy, tolerability and simplicity, low cost, and good adherence to treatment (WHO 2003).

Generic fixed-dose combinations of such regimens are widely regarded as crucial for scaling-up AIDS treatment in developing countries. These treatments improve adherence owing to the fewer daily doses relative to individual formulations. Supply, storage, and distribution are also easier because the range of products is smaller. Generic drugs are generally much cheaper than brand-name formulations. Several generic fixed-dose combinations have been pre-qualified by WHO (WHO 2004) after assessment of manufacturers’ product data (including data for purity of all ingredients, stability of the finished products, and results of in-vivo bioequivalence tests), actual pharmacological composition, and manufacturing practices. However, these formulations are not yet recommended by some of the major donor agencies, such as the US government's multi-billion dollar PEPFAR (President's emergency plan for AIDS relief funding) programme for developing countries (USAID 2004). In addition to political considerations, particularly on the legitimacy and consequences of using generic instead of brand-name drugs, this situation is partly explained by the absence of clinical studies showing the efficacy and tolerability of generic fixed-dose combinations. Quality control of different drug batches is also a difficulty in most developing countries.

The generic fixed-dose combination of nevirapine, stavudine, and lamivudine (Cipla, Mumbai Central, Mumbai, India) is one of the most frequently prescribed treatments in African countries. In Angola, a south west African country with more than 15 million inhabitants, the prevalence of HIV infection is increasing rapidly, with up to 8% of town-dwelling pregnant women infected (UNAIDS 2002). HIV-1 predominates, and HIV-1 groups M, N, and O and many subtypes and circulating recombinant forms co-circulate (Vergne at al 2003). A national antiretroviral access programme has started, most usually based on the generic combination of nevirapine, stavudine, and lamivudine owing to its low price. However the sad fact remains that many AIDS-infected patients in Angola go without any anti-viral treatment.

Use of these combination therapies has dramatically decreased morbidity and mortality rates in HIV-1 infected individuals (Hogg et al. 1997; Cameron et al. 1998; Palella et al. 1998). However, such intensive combination therapies have various drawbacks, such as drug side-effects, the complexity of the therapeutic regime, and the appearance of resistant HIV strains (Carr et al. 1998; Colgrove et al. 1998; Sarmati et al. 2002). So far, it appears that these combination antiretroviral drug regimens must remain in use until a radical curative treatment and HIV-1 vaccine are established. Therefore, therapies based on new principles other than drug treatment would appear to be highly desirable.

Electrical stimulation effects on living cells have been extensively studied since the 1970s (Zimmerman et al. 1974). It has been reported that use of high d.c. voltage pulse application not only induces changes in cellular membrane structure and permeability, but also results in its breakdown (Berg et al. 1984; Powell et al. 1986). Moreover, cells suspended in solution were found to be easily fused by pulses of high d.c. voltage (Zimmerman 1982). On the other hand, pulses of low d.c. voltage are known to regulate cellular proliferation and protein production together with induction of differentiation of various cell types (Kojima et al. 1992; Mie et al. 1996; Aizawa et al. 1999). Furthermore, the effects of electrical stimulation on HeLa cells chronically infected with HIV-1LAI and on uninfected P6 HeLa cells (Tominaga et al. 2003) has been reported, and the possible causes for the significant damage that occurs to infected P6 HeLa/HIV-1LAI cells compared with uninfected control cells has been considered.

In this study we tested the effectiveness of an electrotherapy apparatus (The Bio-Lyfe machine), that delivered an oscillating electrical signal via hand-held probes, on the immunological parameters and viral load in HIV infected patients.

Methods

All participants in the trial were periodically monitored with respect to their clinical features; any side effects were noted. Blood was withdrawn and placed into tubes with ethylenediaminetetraacetic acid (EDTA) as anticoagulant; the plasma was used in tests and some was also stored at -70�C. Laboratory tests, including routine blood tests, and CD4 and CD8 counts, were performed at the Military Hospital. The HIV viral load was determined in the HIV Laboratory, Laboratory Services Branch, of the Military Hospital which used the sensitivity cut-off value of 10 000 HIV copies of RNA per millilitre) (Chiron Diagnostics 1997). All data and clinical records were kept confidential.

In this study, 37 patients with HIV infection were treated with The Bio-Lyfe machine. The treatment period was at least 2 months. Patients were exposed to hand held electrode probes daily for 20 minutes at the highest tolerable current. There were no obvious side effects reported throughout the two months. Compliance was difficult to maintain as it involved patients coming to a medical centre daily. Seven patients were discontinued from the trial due to poor compliance. Thirty HIV infected patients who were not participating in any anti-HIV treatment were randomly chosen from the hospital database and matched with trial patients who were observed for two months.

The CD4 and CD8 counts and HIV viral loads of patients were measured before the trial, at 1 month and at 2 months of treatment with the Bio-Lyfe machine (Tables 1 and 2). These results were compared to test results for the untreated patients. All patients gave their written informed consent.

Results

Raw Data Table 1 Treated Patients:

Month 0 Month 1 Month 2
Viral Viral Viral
ID Sex Age CD4 CD8 Load CD4 CD8 Load CD4 CD8 Load
1 M 23 293 417 41475 313 676 31888 304 711 24852
3 M 27 290 377 131802 330 612 114223 409 785 88574
4 M 25 349 724 86182 312 715 78208 339 730 71556
5 M 32 484 853 276163 445 798 278917 422 751 29885
6 M 41 754 1046 229558 688 943 249003 678 936 228557
7 M 23 453 801 208412 445 815 210234 478 866 195633
8 M 27 670 1040 223524 585 927 150749 526 839 169857
9 M 46 707 958 249882 611 978 176912 587 888 156985
10 M 32 369 1002 203176 452 989 207899 352 785 254877
12 M 25 337 871 138702 415 878 87269 357 857 89663
13 M 27 450 759 110019 437 811 105882 411 872 91721
14 M 33 240 651 60737 398 973 28911 408 957 26888
15 M 19 601 1644 297874 583 1534 311092 651 1452 300922
16 M 22 539 712 110183 525 1023 87999 455 1066 79639
17 M 25 270 435 28031 336 672 9021 429 639 7058
18 M 27 246 733 22660 259 755 27898 211 781 22844
20 M 29 321 560 11144 333 829 11453 308 800 18522
21 F 21 370 391 173178 392 722 121022 502 967 59633
22 M 28 345 992 14519 333 947 17900 329 788 25983
23 M 30 171 510 209796 199 615 207678 251 611 298667
24 M 26 680 1219 17131 612 1121 16752 482 899 59633
25 M 22 163 258 737148 172 427 547288 144 389 685214
26 M 19 298 505 98813 323 504 102007 389 585 73695
27 M 18 566 1447 27960 535 1399 33872 524 1189 49685
28 F 39 227 489 398022 254 500 358966 233 496 211589
29 M 42 188 412 678876 189 397 402587 161 385 54833
Patient 2, 11 and 19 discontinued.

Table 2 Untreated patients:

Month 0 Month 1 Month 2
Viral Viral Viral
ID Sex Age CD4 CD8 Load CD4 CD8 Load CD4 CD8 Load
1 M 26 379 795 87177 311 660 89155 308 489 163588
3 M 19 561 1338 29227 544 1125 39565 452 1159 36852
4 M 42 371 1393 38237 312 915 71589 215 689 158009
5 M 38 194 365 349193 204 411 334568 261 497 193578
6 M 30 635 1484 57671 678 1358 65487 587 1258 74269
7 M 46 261 671 204635 258 615 219284 199 422 339658
8 M 40 565 1246 32381 572 968 29567 518 702 56892
9 M 19 319 1041 102341 288 777 198557 333 693 236952
10 M 19 525 1452 34802 535 1544 20568 439 1423 38978
12 M 37 549 1531 38977 515 1422 37552 482 1112 155369
13 M 27 602 1317 23262 438 891 43255 339 672 252361
14 M 29 594 1156 33517 598 1211 29658 502 1005 30251
15 M 29 466 1122 116590 483 1534 135985 389 1025 189752
16 M 20 212 635 30907 241 702 28564 149 583 96352
17 M 23 219 637 154793 215 672 155888 226 545 162358
18 M 19 519 1285 74609 485 1158 95685 389 901 142365
20 M 40 193 497 183090 202 544 175882 159 452 196358
21 F 22 451 965 29391 435 789 37551 242 498 189635
22 M 23 307 694 33068 333 752 31588 242 593 63258
23 M 22 171 438 172352 170 411 218678 193 452 152981
24 M 38 503 1101 17579 544 1004 16884 539 996 18596
25 M 37 508 1040 62644 333 775 221479 406 779 15639
26 M 45 242 640 200383 244 581 200875 269 621 185269
27 M 22 158 335 275711 148 347 338547 158 385 25689
28 F 25 145 440 394090 140 451 401585 129 421 489366
29 M 25 535 1114 26904 517 1220 27568 506 1107 36952
30 M 27 412 811 12459 388 795 16587 376 763 20562

Statistics

The purpose of the study was to study the use of electrotherapy on HIV-1 patients. There were 67 patients enrolled; 53 of these completed a 2-month period of involvement in the study. Of these, 26 received a daily oscillating electrical signal treatment over a period of at least 2 months, while 27 (the controls) did not.

The patients to be treated were hand-selected by the researcher from the hospital database, with the intention of selecting a representative range of values for the outcomes of interest, CD4 and CD8 lymphocyte count and viral load. The controls were randomly selected from 80 of the remaining patients on the database who were deemed to be similar in age and gender to those selected to be treated, though they were not individually matched. Patients were excluded from the study if they were already receiving an alternative HIV treatment. All 53 patients were monitored over a 2 month period, and measurements of CD4 count, CD8 count and viral load were made at 0, 1 and 2 months.

In this report, we present analyses to address the question:

• Is there a difference in the CD4 count, CD8 count and viral load between patients who received electrotherapy and those who did not?

Method

The data were provided in two files, one for treated and one for untreated patients. The following information was provided for each patient: gender, age and measurements for the three outcomes at each of three time points.

Since the selection of patients for treatment was not a standard random selection, there could have been bias in the selection process. Although it is difficult to dispute this point, initial measurements for the treated and untreated groups were compared, which would be expected to be similar if there was no selection bias.

For each outcome, the main variable of interest was the change between the first measurement and the 2-month measurement, as a percentage of the first measurement. The percentage change was of interest because there was a large amount of variability in the measurements between patients. The two groups were compared separately for the three variables of interest. As the data did not appear to be normally distributed, nonparametric tests were used for these comparisons. The particular test used in each case was the Mood test, which makes no assumptions about the distributions of the two outcomes. Other, less conservative tests could have been used and did, in fact, indicate more significant differences between the groups, but these relied upon assumptions which did not appear to be valid when visual representations of the data were considered.

Results

1. Comparing initial values for the two groups

The gender representation in the groups was almost identical (2 females in each) and the mean ages were not statistically significantly different between the two groups (29 years for the untreated compared with 28 years for the treated group).

Dotplots of the initial values for each of the three outcomes are given below, along with a table of descriptive statistics for these outcomes. None of these differed significantly between the groups, although there was a greater spread of values for viral load in the treated group, probably due to the way that these patients were selected. This difference may have implications for the final results.

Dotplots of initial CD4 and CD8 counts and viral load

No Yes

CD4 count

160 240 320 400 480 560 640 720

Treated

No Yes

CD8 count

360 540 720 900 1080 1260 1440 1620

No Yes

0 100 200 300 400 500 600 700

viral load (X1000)
Standard
Outcome Group n Mean deviation Minimum Median Maximum
CD4 count Untreated Treated 27 26 392 399 161 176 145 163 412 347 635 754
CD8 count Untreated Treated 27 26 946 762 373 343 335 258 1040 728 2531 1644
Viral load (X103) Untreated Treated 27 26 104 184 105 185 12 11 58 135 394 737

2. Comparing the treatment effect over time

Below are the results for the three outcomes. In each case, the same information has been provided: a dot plot comparing the percentage changes for the two groups, summary information of the percentage change for the two groups and the results of the Mood test comparing the median percentage change for the two groups. For each test, a P-value has been given, which is the probability of observing the difference in medians that we have or a more extreme difference, assuming there was no underlying difference. Obviously a small P-value is evidence of a difference between the groups and a P-value is usually considered to be statistically significant if it is below 0.05.

CD4 lymphocyte count

Dotplot of percentage change in CD4

-48 -32 -16 % change in CD4 160 32 48 64
Summary statistics for percentage change Group n Mean Standard deviation Untreated 27 –12.4 18.0 Treated 26 5.0 25.9 Minimum –46.3 –29.1 Median –15.5 –4.3 Maximum 34.5 70.0

Results of the Mood test

Difference between medians (untreated minus treated) = –11.2 95% confidence interval for difference between medians: (-22.6, 0.4)

P-value = 0.07

CD8 lymphocyte count

Dotplot of percentage change in CD8

Treated

No Yes

-30 0 % change in CD8 6030 90 120 150
Summary statistics for percentage change Group n Mean Standard deviation Untreated 27 –16.7 20.4 Treated 26 18.3 42.3 Minimum –50.5 –26.3 Median –13.4 4.0 Maximum 36.2 147.3

Results of the Mood test

Difference between medians (untreated minus treated) = –17.4 95% confidence interval for difference between medians: (-41.1, -2.1)

P-value = 0.004

HIV viral load

Dotplot of percentage change in viral load

% change in viral load

Summary statistics for percentage change

Standard
Group n Mean deviation Minimum Median Maximum
Untreated 27 108.6 220.2 –90.7 37.4 984.9
Treated 26 –5.9 69.1 –91.9 –20.5 248.1

Results of the Mood test

Difference between medians (untreated minus treated) = 57.9 95% confidence interval for difference between medians: (18, 109)

P-value < 0.001

Conclusion

Our tests found that there was a statistically significant difference between the two groups for the outcomes of CD8 count and viral load. The treated group had:

  • a small positive median percentage change in CD8 lymphocyte count, compared to a large negative median percentage change for the untreated group;
  • and a large negative median percentage change in HIV viral load, compared to a large positive median percentage change for the untreated group.

In these results, there was a consistently better outcome in the treated group, although the difference in median CD4 count was not significant.

There are important reasons why it is difficult to infer from these results that the treatment generally causes a beneficial effect.

As the patients selected to receive the treatment were hand-chosen in order to provide a good range of values (while the controls were a simple random sample), differences between the groups may be the result of more than just the electrotherapy. For example, viral load was generally larger in the treatment group and it could be argued that a larger initial value generally results in a larger decrease in viral load over time.

Also, even though the patients were selected from the same population, the specific nature of this population does have implications for any inference that can be made. It is not possible to tell how patients in a military hospital may differ from the general population, but it is highly unlikely that this population is representative of the wider HIV-1 population of Angola.

Graph of HIV Viral Load Changes Over 2 Months for Individual Patients in the Treated Group

HIV Africa Treated Patients

Viral load

300

250

200

150

100

Series1

50

0

-50

-100

-150

Patient number

Graph of Change in HIV Viral Load Over 2 Months for Individual Patients in the Untreated Group

HIV Controls Viral Load

% Viral Load

1200

1000

800

600

Series1

400

200

0

-200

Patients

CD4 + CD8 Graph – Summary Results - % Change over 2 months

HIV Africa 2005

Group

Viral Load Changes Graph – Summary Results

HIV Electrotherapy Study Africa 2005

Patient Groups

Discussion

There is a positive effect of treatment with The Bio-Lyfe electrotherapy machine on CD8 cell count followed by and a negative significant effect on the viral load viral load a positive non significant effect on CD4 count in many of the HIV-infected patients. Large individual variations in response were also observed. We have previously observed positive effects in CD8 cell counts in various cancer patients and in three HIV patients (unpublished data).

The multi wave oscillating device provides an electrical signal to two hand held electrodes that boost CD8 immune system cell numbers.

By boosting these CD8 cells the viral load is reduced, and the treated patients experience an improvement in the quality of life and an extension of life.

Research at Duke University has shown that high levels of CD8 cells have a positive effect on prognosis in AIDS-infected patients. The proposed mechanism of action is that in asymptomatic patients, CD8 cells can identify infected CD4 cells, latch onto them, and release compounds that cause the infected cell to burst, killing it.

CD8 cells also possess a non-cytolytic weapon as well, and this stops HIV replication.

Specifically Tomaras and Greenberg’s experiments (2000) at Duke University show that CD8 cells affect the virus after it has already entered the CD4 cell, which is different to the way beta-chemokines work. The CD8 cells stopped HIV from hijacking the CD4 cell’s genetic machinery to reproduce itself.

Discussion Of Other Relevant Studies

Immunologic characteristics have previously been studied in 103 patients with multiple myeloma, acute leukemia, chronic lymphocytic leukemia and non-Hodgkin's disease following in vitro exposure of blood to a low-intensity static field (SF) and alternating field (AF) or pulsating magnetic field (PF) (Bessmel'tsev 2001). In this study of multiple myeloma, a 30 minute exposure had a positive effect on expression of tumor cells and T-cell markers and stimulated the regulatory function of T-lymphocytes. With SF-AF and PF application alternating, the expression of both +CD3 and +CD4 and the +CD3/+CD4 ratio increased suggesting the lowering of immunological deficiency. In acute leukemia, a combined application of the magnetic fields had an effect on the helper activity of the T-lymphocyte sub-population. The phagocytic activity of leukocytes increased significantly while their digestive ability rose to a moderate degree.

The effect of leukocyte subsets, total leukocyte isolates or full blood samples subjected to medium-strength square-wave electric impulses (100 V/cm field force, 5 ms duration) has also been reported (Filipic 2000). On the surface of the leukocytes, the expressions of several markers (CD3, CD4, CD8, CD11a, CD11b and ICAM-1) were determined in order to study the influence of pulsed ionic currents on different aspects of the cellular immune response. Large individual differences were also observed in this study among randomly chosen healthy donors, both in the initial expression rate and in the response patterns of different antigens. As a general conclusion, it can be stated that electric impulses with the above parameters activate the state of immune response alertness of human leukocytes. Elevation in the activities of several enzymes, such as lactate dehydrogenase and superoxide dismutase in the serum in response to electric impulses suggested an antiviral and immune activated condition. It was concluded that ex vivo blood treatment with medium-strength electric impulses seems to be a promising adjuvant course for the establishment of acute immune potentiation and an antiviral state in patients undergoing dialysis treatment.

The effects of uncontrollable and controllable electric shocks on the immune system in rats (the proportion of CD4+, CD8+ or CD25+ T lymphocytes to total lymphocytes was measured in the peripheral blood, spleen, and thymus) have also been observed (Nakata 1996). The rats were given either controllable shocks, identical uncontrollable shocks, or no shocks, and then small shocks 24 h later (reinstating shocks). The proportion of CD4+ T lymphocytes relative to total lymphocytes in both the peripheral blood and spleen of uncontrollable rats (URs) was significantly smaller than was found in no-shock rats (NRs). Similarly, the proportion of CD4+ T lymphocytes in the thymus of controllable rats (CRs) was significantly smaller than in NRs. In contrast, the proportion of CD8+ T lymphocytes in the thymus of URs was significantly larger than in NRs and CRs. The CD4+ to CD8+ T lymphocyte ratios (CD4+/CD8+ ratios) in the peripheral blood, spleen and thymus of URs were significantly smaller than in NRs; also, the ratios in the peripheral blood and spleen of URs were significantly smaller than in CRs. The white blood cell (WBC) count of URs was significantly smaller than those of NRs and CRs, and the WBC count of CRs was significantly smaller than those of NRs and CRs, and the WBC count of CRs was significantly smaller than that of NRs. These results suggest that decreases in CD4+ T lymphocytes (and/or an increase of CD8+ T lymphocytes) in the peripheral blood, spleen, and thymus are caused by uncontrollable stress followed by a reinstating stress condition, leading to the decrease of WBC in the peripheral blood and decreases in the CD4+/CD8+ ratios in these tissues.

Electrical stimulation can also affect neuropathies in nerves and circulating neurotransmitter levels (Herzberg 1995). This indirectly can affect the immune system (Lechin 2002). To study the possible mechanism by which peripheral nerves mediate immune responses in target tissues, electrical stimulation of the sciatic nerve was combined with subcutaneous microdialysis of the hind paw (Herzberg 1995). Following unilateral stimulation of the sciatic nerve, an ipsilateral rise in substance P and a bilateral rise in VIP levels were observed in dialysate samples from experimental vs control animals. Electrical stimulation of the sciatic nerve induced a marked hyperemia and swelling of the ipsilateral paw. Quantitative immunocytochemical analysis of paraffin-embedded sections of the hind foot pads demonstrated T lymphocyte migration ipsilateral to the stimulated nerve. These findings suggest that peripheral nerves can directly modulate local immune and inflammatory responses.

It has been shown in previous studies that cell poration (i.e. reversible permeabilization of cell membrane) and cell fusion can be induced by applying a pulse (or pulses) of high-intensity DC (direct current) electric field (Chang 1989). The same group also suggested that such electro-poration or electro-fusion can also be accomplished by using an oscillating electric field. The DC field relies solely on the dielectric breakdown of the cell membrane to induce cell fusion. The oscillating field, on the other hand, can produce not only a dielectric breakdown, but also a sonicating motion in the membrane that could result in a structural fatigue. Thus, a combination of a DC field and an oscillating field is expected to enhance the efficiency of cell poration and cell fusion. Pulses of high-intensity, DC-shifted RF (radio frequency) electric field were used to induce a highly efficient cell poration and cell fusion on human red blood cells and on a fibroblast cell line.

Another study demonstrated a safe and effective way to introduce exogenous genes into cells, a new method of electroporation which uses a radio-frequency (RF) electric field to permeabilize the cell membrane has been uncovered (Chang 1991). This RF method has several advantages over the conventional electroporation method which uses a direct current (DC) field. It was shown that the RF electroporation method can be used to introduce marker genes into a wide variety of cell lines and was able to increase substantially the efficiency of gene transfection. Interestingly, the transfection efficiency was shown to be affected by a number of factors, including cell type, field strength, pulse protocol and medium buffer. Because of its wide range of applications, high transfection efficiency and lack of harmful side-effect, the RF electroporation method would be particularly useful for introducing genes into human cells for gene therapy.

Morphological changes of HIV infected cells following application of low electrical potential were induced due to a decrease in the plasma membrane fluidity and deformation of cytoskeletal structure including F-actin (Aizawa et al. 1999; Yaoita et al. 1989). It has also been suggested (Kumagia 2004) that the swelling and breakdown of cell membrane due to application of electrical potential might have occurred due to an increase in the adhesion area between the positively charged ITO electrode and the negatively charged cells after electrical stimulation. Several other factors possibly contribute to the higher cellular damage due to electrical stimulation in P6 HeLa/HIV1LAI cells than in P6 HeLa cells. It was considered that the most important factor to be the large surface area of chronically HIV-1 infected cells due to the budding of HIV-1 particles at the plasma membrane, which is not observed in uninfected cells (Barré-Sinoussi et al. 1983; Chrystie and Almeida 1988). In other words, budding of HIV-1 would increase the negative charge of infected cells, and hence lead to increased reception of the effect of electrical stimulation. Another factor would be that some oxidized species might have developed due to the electrochemical reaction at the ITO electrode surface in the supernatant of P6 HeLa/HIV-1LAI cells. Collectively, it has been shown (Kumagai 2004) that the difference in sensitivity to electrical stimulation between uninfected cells and cells chronically infected with HIV-1 could be useful not only for the detailed elucidation of HIV control mechanisms but also for the development of new therapies.

Summary - In Vivo Pulsed High Potential Electrotherapy

Pulsed high potential electrotherapy technology appears to offer a novel approach to treating diseases related to immunosuppression, such as infection with HIV.

The technique involves the use of brief high potential spikes which disrupt the skin’s dielectric barrier and allow specific electrical potentials to penetrate into the body’s tissues. These potentials carry specific waveforms and frequencies which can have positive effects on the healthy function of a range of tissues in the body. In addition these high electrical potentials can disrupt the weakened membranes of infected cells, causing cell death. These potential also appear to have a specific anti-microbial effect. It is thought that electromechanical and electrostatic stresses are induced by these high voltage pulses at multiple audio and radio frequencies.

The electromechanical stresses, cell electroporation, and ion exchange processes induced by high voltage DC pulses are well documented in vitro. These processes may act to both destroy infected cells and to stimulate the immune system. (Bryant 1987, Chang 1989, Dao-Sheng 1990, Dimitrov 1990)

Inactivation and destruction of bacteria by electric treatment methods is well established in vitro. In addition pulsed high voltage electrotherapy at multiple frequencies appears to inactivate pathogenic micro-organisms in vivo. (Allen 1966, Hamilton 1967, Gillibrand 1967)

In addition high potential electrotherapy is showing promise in treating inflammatory conditions (Johnson 2004), and in treating cancer – both as a direct cytotoxic agent (Fedorowski 2004, Hernandez-Bule 2004, Kirson 2004, Pinero 1997) and also by means of enhancing the efficacy of chemotherapy and radiotherapy through electroporation effects on the membranes of cancer cells (Ito 2001, Engstrom 2001, Gray 2000, Li 2003, Rabussay 2002, Rols 2000, Sun 2003).

Conclusion

There are multiple studies in the literature suggesting that electrical stimulation can induce positive immunological changes both in vivo and in vitro, and negatively effect HIV-infected cells with no effect on non-infected cells. The Bio-Lyfe electrotherapy apparatus seems to have an obvious positive effect on HIV-infected individuals in this pilot study.

However it’s mechanism of action for inducing these changes is currently unclear. Further more detailed and larger trials on the efficacy and safety of The Bio-Lyfe machine are warranted, as are studies into the possible mechanism of action of The Bio- Lyfe machine. It is of note that The Bio-Lyfe machine appears to be a cheap, safe and easy to use treatment system. It may even be appropriate for home use. The Bio-Lyfe machine looks to be a real possibility to add to the very limited arsenal of weapons to fight the current global HIV epidemic. Further studies are therefore warranted.

Acknowledgements:

Statistics were prepared by Sandy Clarke BSc(Hons) GStat, Statistical Consulting Centre, University of Melbourne.

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Bio-Lyfe Machine

Index

Investigation of the effect of a multi wave oscillator in the treatment of adult HIV/AIDS patients: ........................................................................................................ 1 Abstract................................................................................................................... 1 Introduction............................................................................................................ 1 Methods................................................................................................................... 3 Results..................................................................................................................... 5 Raw Data......................................................................................................................... 5 Table 1 Treated Patients: ................................................................................................ 5 Table 2 Untreated patients: ............................................................................................. 6 Statistics .............................................................................................................................. 7 Method ........................................................................................................................ 7

1. Comparing initial values for the two groups....................................................... 8 CD4 lymphocyte count ............................................................................................... 9 CD8 lymphocyte count ............................................................................................. 10 Graph of HIV Viral Load Changes Over 2 Months for Individual Patients in the Treated Group ............................................................................................................... 13 Graph of Change in HIV Viral Load Over 2 Months for Individual Patients in the Untreated Group............................................................................................................ 14 CD4 + CD8 Graph – Summary Results - % Change over 2 months ............................ 15 Viral Load Changes Graph – Summary Results........................................................... 16 Discussion............................................................................................................. 17 Discussion Of Other Relevant Studies.................................................................. 17 Summary - In Vivo Pulsed High Potential Electrotherapy................................... 20 Conclusion ............................................................................................................ 20 References............................................................................................................. 21 Index ..................................................................................................................... 26