John O. Naim

Photomodulation of Oxidative Metabolism and Electron Chain Enzymes in Rat Liver Mitochondria

Abstract— Low‐level laser irradiation has been applied in a variety of laboratory studies and clinical trials for photobiostimulation over the last three decades. Considerable skepticism exists regarding the concept of photostimulation within the medical community. One of the major difficulties with photoirradiation research is that it lacks experimentally supportable mechanisms for the alleged photobiostimulatory effects. This study was undertaken to determine whether oxidative metabolism and electron chain enzymes in rat liver mitochondria can be modulated by photoirradiation. Oxygen consumption, phosphate potential, and energy charge of rat liver mitochondria were determined following photoirradiation. Activities of mitochondrial enzymes were analyzed to assess the specific enzymes that are directly involved with the photostimulatory process. An argon‐dye laser at a wavelength of 660 nm and at a power density of 10 mW/cm2 was used as a photon source. Photoirradiation significantly increased oxygen consumption (0.6 J/cm2 and 1.2 J/cm2, P < 0.05), phosphate potential, and the energy charge (1.8 J/cm2 and 2.4 J/cm2, P < 0.05) of rat liver mitochondria and enhanced the activities of NADH: ubiquinone oxidoreductase, ubiquinol: ferricytochrome C oxidoreductase and ferrocytochrome C: oxygen oxidoreductase (0.6 J/cm2, 1.2 J/cm2, 2.4 J/cm2 and 4.8 J/cm2, P < 0.05). The activities of succinate ubiquinone oxidoreductase, ATPase, and lactate dehydrogenase were not affected by photoirradiation.

THE EFFECT OF LASER IRRADIATION ON THE RELEASE OF bFGF FROM 3T3 FIBROBLASTS

Studies have shown that low‐level laser irradiation increases the proliferation of fibroblasts in cell culture. The mechanism of action is unknown. Basic fibroblast growth factor (bFGF) is a multifunctional polypeptide that has been detected in most tissues and which supports cell proliferation and differentiation. The purpose of this study was to determine whether laser irradiation (660 nm) can stimulate production of bFGF from fibroblast cells in cell culture. Our study showed that fibroblasts irradiated with laser energy at 2.16 J/cm2 demonstrated increased cell proliferation and enhanced production of bFGF, whereas fibroblasts irradiated with laser energy at 3.24 J/cm2 neither demonstrated increased cell proliferation or an enhanced release of bFGF as compared to the control group. These results provide direct evidence that the proliferation of fibroblasts as a result of stimulation by low level laser irradiation may be associated with the autocrine production of bFGF from fibroblasts.

The Adjuvant Effect of Silicone-Gel on Antibody Formation in Rats

The extent of immunological adjuvancy of silicone-gel, from mammary implants, up to now, has not been determined definitively. This study compares the immune potentiation effects of silicone-gel with that of Freunds adjuvant, using bovine serum albumin (BSA) as the test antigen in rats. Sixty, 250 gr., male Sprague Dawley rats were divided into six groups: I- phosphate buffered saline (PBS) only, II- silicone oil (Dow Corning Medical Grade 360 liquid silicone), III- 50% silicone-gel (McGhan Medical Corp.- mammary implant) in silicone oil, IV- complete Freunds adjuvant (CFA), V- incomplete Freunds adjuvant (IFA), and VI- 50% silicone oil in IFA. Each adjuvant was mixed or emulsified with an equal volume of 50 micrograms of BSA in 150 microliters of PBS. Each immunization was given intramuscularly in a single injection. Cardiac puncture test bleeds were taken at 12, 22, 40 and 56 days post immunization and the serum anti-BSA-antibody was measured by ELISA. The results indicate that silicone-gel is a potent immunological adjuvant, compared to both CFA and IFA. Silicone oil alone is not as potent as adjuvant and seems to inhibit the immune response when mixed with IFA. There thus appears to be a distinct possibility that silicone-gel may also be able to mediate an auto-immune reaction.

IMPACT OF DIFFERENT ASBESTOS SPECIES AND OTHER MINERAL PARTICLES ON PULMONARY PATHOGENESIS

Factors that are potentially important in the pulmonary pathogenesis of asbestos and other mineral particles are: 1) morphology, 2) Fe-content, 3) solubility under intraphagosomal conditions, 4) value and sign of the surface potential of the particle, 5) hydrophobicity or hydrophilicity, 6) capacity to activate phagocytic leukocytes, and 7) duration of exposure to the particles. The order of importance of these factors in causing severe or fatal pulmonary pathogenicity is estimated to be: 1 > 3 > 7 > 6 ≫ 5 > 4 > 2. The order of pathogenicity of the minerals is estimated as: amphibole asbestos: crocidolite, tremolite, amosite > erionite > serpentine asbestos: chrysotile > talc > silica > simple metal oxides. Particle length, duration of exposure to the particles, and pre-treatment of the particles may however enhance the pathogenic potential of any of the lower-ranked particles.

The effect of site and technique of splenic tissue reimplantation on pneumococcal clearance from the blood.

The technique and site of reimplantation of splenic tissue influences survival of laboratory animals following intravenous injection of pneumococci. Splenic tissue was prepared by slicing, mincing, or grating the spleen. The tissue was placed subcutaneously, intraperitoneally, retroperitoneally, or in an omental pouch. This study was designed to determine the rate of pneumococcal clearance from the blood stream 16 weeks following splenic reimplantation by four different methods. All animals were challenged with an intravenous 1 mL bolus containing 10(7) bacteria. The New Zealand white rabbits were divided into six groups: intact spleen; splenectomized; spleen slices in an omental pouch; minced spleen in an omental pouch; splenic tissue implanted subcutaneously; and bits of spleen dropped into the peritoneal cavity. Animals with an intact spleen and those with spleen slices implanted into an omental pouch cleared bacteria during the first hour and all bacteria had disappeared at three hours. Bacteremia persisted longer than three hours in the other groups. Splenic tissue had regenerated in all animals with omental pouch implants, in four of six with minced spleen dropped into the peritoneal cavity but in only one with a subcutaneous implant. Reimplanted splenic tissue clears pneumococci from the blood stream best when thin slices of spleen are placed in an omental pouch. This technique also assures successful regeneration of splenic tissue.

Beneficial effects following carbon dioxide laser excision on experimental neuroblastoma

Treatment of neuroblastoma in children consists of primary excision with adjuvant radiation and chemotherapy. When the tumor invades surrounding structures that cannot be safely excised or when distant metastasis is present, the patient has a poor prognosis. Because the CO2 laser can be used to excise malignant tumors without seeding the surrounding tissue and because the defocused beam can vaporize malignant cells, we compared partial scalpel excision and partial laser excision of C1300 murine neuroblastoma to the growth rate of residual tumor. In 25 mice, 75% of the tumor was excised with a scalpel, and in another 25, the same percentage was excised with the CO2 laser (10 W). CO2 laser excision significantly decreases the growth of residual neuroblastoma (P less than .01). However, the effect appears to be a function of increased tumor immunogenicity after laser excision rather than the increased tumor kill. We conclude that CO2 laser excision of neuroblastoma may prove to be superior to scalpel excision for primary surgical treatment of neuroblastoma.

Induction of Type II Collagen Arthritis in the DA Rat Using Silicone Gel as Adjuvant

The Dark Agouti (DA) rat has been shown recently to have a high susceptibility for developing arthritis when challenged with either heterologous or homologous collagen II mixed with mineral oil, or with mineral oil challenge alone. This study determined the arthritogenic potential of silicone gel by either mixing it with bovine collagen II (BII) or by injecting silicone gel alone in DA rats. The incidence of collagen induced arthritis was as follows: PBS group- 0/10, silicone gel group- 4/10, and IFA group- 8/9. Anti-BII antibodies were formed in most of the rats treated with either silicone gel or IFA and these groups of rats showed a positive DTH reaction. The PBS treated rats were negative for both anti-BII antibodies and DTH reaction. The incidence of arthritis formation in rats injected with silicone gel alone was 0/10, while the IFA injected rats showed an incidence of 8/10. Silicone gel taken from a commercial breast implant thus is capable of mediating collagen induced arthritis in the DA rat. However, silicone gel alone does not appear to be arthritogenic.