Immunologic and hematopoietic alterations by 2,450-MHz electromagnetic radiation

A biphasic modulation of responsiveness of spleen lymphocytes to mitogens was observed in mice exposed to 2,450-MHz radiation at power densities of 5–15 mW/cm² over various periods ranging between one and 17 days. This modulated phenomenon may be explained on the basis of 1) suppression of lymphocyte response by microwave-activated macrophages which persists throughout the entire course of radiation, and 2) concurrent progressive direct stimulation of lymphocytes which culminates around day 9 of exposure. Tumor cytotoxicity of killer lymphocytes from mice exposed to five or nine days of radiation did not appear different from sham controls. The highly proliferative hematopoietic marrow cells were sensitive to microwave radiation. Nine days of exposure to radiation (15 mW/cm²) reduced the colonyforming units of myeloid and erythroid series by 50%. This observation may offer a new and more sensitive assay for studying biological effects of electromagnetic radiation.     

Effect of nonionizing radiation on the purkinje cells of the rat cerebellum

In one experiment, Sprague Dawley rats (16–21 days of gestation) and their offspring were exposed to 100-MHz (CW) electromagnetic radiation at 46 mW/cm² (SAR 2.77 mW/g) for 4 h/day for 97 days. In another experiment, the pregnant rats were irradiated daily from 17 to 21 days of gestation with 2450-MHz (CW) microwaves at 10 mW/cm² (SAR 2 mW/g) for 21 h/day. In a third experiment, 6-day-old rat pups were irradiated 7 h/day for five days with 2450-MHz radiation at 10 mW/cm². Equal numbers of animals were sham irradiated in each group. Quantitative studies of Purkinje cells showed a significant and irreversible decrease in rats irradiated during fetal or fetal and early postnatal life. In animals exposed postnatally, and euthanized immediately after irradiation, significant decrease in the relative number of Purkinje cells was apparent. However, restoration apparently occurred after forty days of recovery.     

Effects of Microwave Exposure at Various Power Densities on Mitochondrial Marker Enzymes in Mouse Brains

Four groups of C₅₇BL mice were irradiated with 3 GHz pulse (PW) microwaves for 3 hours at incident power densities of 0.1, 0.5, 1 and 5 mW/cm² respectively. The amount of mitochondria1 marker enzymes succinate dehydrogenase (SDH) and monoamine oxidase (MAO) in the hypothalamus and hippocampus were determined by microspectrophotometry. SDH and MA0 in the irradiated groups (except 0.1 mW/cm²) were significantly lower compared to the control group (p < 0.01). The lowest level occurred in the 5 mW/cm² group. The threshold level was 0.5 mW/cm². To compare the effects of PW with continuous wave (CW) exposure, two experimental groups were exosed to 2.45 GHz, using CW; the enzymes were decreased only in the 5 mW/cm² group. The results show that PW radiation is more effective then CW radiation in decreasing SDH and MA0 levels.     

Effects of 2.45-GHz microwave radiation on embryonic quail hearts

Although exposure to nonionizing electromagnetic radiation has been reported to cause a variety of systemic alterations during embryonic development, there are few reports of the induction of specific physiologic or morphologic changes in the myocardium. This study was designed to examine the effects of microwave radiation on cardiogenesis in Japanese quail embryos exposed during the first eight days of development to 2.45-GHz continuous-wave microwaves at power densities of 5 or 20 mW/cm². The specific absorption rates were 4.0 and 16.2 mW/g, respectively. The ambient temperature for each exposure was set to maintain the embryonated eggs at 37.5 °C. This did not preclude thermal gradients in the irradiated embryos since microwaves may not be uniformly absorbed. The test exposure levels did not induce changes in either the morphology of the embryonic heart or the ultrastructure of the myocardial cells. Analysis of the enzymatic activities of lactate dehydrogenase, glutamic oxaloacetic transaminase, and creatine phosphokinase failed to reveal any statistically significant differences between the nonexposed controls and those groups exposed to either 5 or 20 mW/cm². The data indicate that 2.45-GHz microwave radiation at 5 or 20 mW/cm² has no effect on the measured variables of the Japanese quail myocardium exposed during the first eight days of development.     

Alteration of life span of mice chronically exposed to 2.45 GHz CW microwaves

Female CD 1 mice were exposed from the thirty-fifth day of age for the remainder of their lives to 2.45 GHz, CW-microwave radiation at a power density of 3 or 10 m W/cm² (SAR = 2.0 or 6.8 W/kg). Exposures took place 1 h/day, 5 day/week in an anechoic chamber at an ambient temperature of 22 °C and a relative humidity of 50%. There were 25 animals in each exposure group, and an equal number of controls were concurrently sham exposed. The average life span of animals exposed at 10 mW/cm² was significantly shorter than that of sham-exposed controls (572 days vs. 706 days; P = .049; truncation >20%). In contrast, the average lifespan of the animals exposed at 3 mW/cm² was slightly, but not significantly, longer (738 days) than that of controls (706 days). © 1994 Wiley-Liss, Inc.

1 This article is a US Government work and, as such, is in the public domain in the United States of America.

     

In vitro microwave effects on human neutrophil precursor cells (CFU-C)

Human marrow cells were irradiated with 2450-MHz CW microwaves in a fluid-filled waveguide irradiation system. Cell exposure was conducted by placing a marrow cell suspension in 20-μl glass microcapillary tubes that were positioned in the exposure chamber, and irradiated at power densities from 31 to 1,000 mW/cm² (with corresponding specific absorption rates of 62 to 2,000 mW/g) for 15 minutes. The temperature of the sample was maintained at a fixed point. Sham-irradiated (SI) and microwave-irradiated (MWI) cells were cultured in a methylcellulose culture system for neutrophil colony proliferation. There was no reduction in neutrophil colony number on days 6–7 or 12–14 in cells exposed at 31 or 62 mW/cm², but as the power density was increased to 1,000 mW/cm², there was a reduction in colony number of MWI cells compared with SI cells. The microwave interaction with the human neutrophil colony-forming cells was apparently not related to temperature rise, or to the state of cell cycle, and was irreversible.     

Effect of low-power red light laser irradiation on the viability of human skin fibroblast

Human skin fibroblast monolayers (S-126 cell line) were exposed to laser radiation (wavelength 670 nm, power density 40 mW/cm²). The energy densities were 2 J/cm² and 12 J/cm², respectively, and the irradiation was carried out at a temperature of 22°C. For fibroblast viability evaluation, the colorimetric assay (conversion of thiazolyl blue to formazan) was used. The experiments were carried out at 37°C, in the presence of 5% CO₂, and at different time periods of incubation after irradiation (2, 4, 8 h and 1, 2, 3, 4, 5 days). The results indicated that there was a certain stimulating effect on the long-term proliferation of skin fibroblasts and that the stimulation proceeded in two stages, the first one 2 h and the second one 3 days post-irradiation. Received 7 January 1998 / Accepted in revised form: 11 June 1998     

The in vivo effects of 2.45 GHz microwave radiation on rabbit serum components and sleeping times

Summary An investigation was conducted to determine the effects of relatively low power density microwave exposures on various serum components of the Dutch rabbit. Both continuous wave and pulsed mode exposures at 2.45 GHz were used at power densities of 25, 10 and 5 mW/cm². Studies of 10 serum components were performed. Additional studies were conducted on changes in sleeping times of pentobarbital-sedated rabbits at various power densities. Gross and histopathological examinations were performed on representative samples of animals.Changes in the blood chemistry of irradiated animals were consistent with a dose-dependent response to a non-specific thermal stress at all power densities used. Observed physiological response, as well as rectal temperature measurements, indicated that the thermoregulatory capability of the rabbits was sufficient to compensate for the thermal burden at 5 and 10 mW/cm², but could be overridden by a 2 h exposure at 25 mW/cm². Pathology findings included a mild, repairable nephrosis in animals exposed at a power density of 25 mW/cm².A further investigation of analeptic effects at power densities varying from 5 mW/cm² to 50 mW/cm² resulted in a statistically significant decrease in sleeping times, apparently proportional to power density below 15 mW/cm².This research was partially supported by the US Army Medical Research and Development Command, Contract No. DADA17-72-C-2144. (The views expressed are those of the authors and do not necessarily reflect those of the Department of the Army)     

Alteration of circulating antibody response of mice exposed to 9-GHz pulsed microwaves

A significant increase was observed in the circulating antibody titers of mice exposed to 9-GHz pulsed microwaves at an average power density of 10 mW/ cm², two hours per day for five days compared with sham-irradiated animals. The mice were previously immunized with type III pneumococcal polysaccharide. Following irradiation, a portion of the immunized animals were challenged with virulent ^{Streptococcus pneumoniae}, type III. Ten days after challenge, mortality was essentially the same in the two groups, but during the ten day period, there was a noticeable increase in the survival time of the irradiated animals compared with the sham-irradiated animals, suggesting that the increased circulating antibody response afforded some degree of temporary protection to the animals.     

Induction of apoptosis coupled to endoplasmic reticulum stress in human prostate cancer cells by n-butylidenephthalide

Background

N-butylidenephthalide (BP) exhibits antitumor effect in a variety of cancer cell lines. The objective of this study was to obtain additional insights into the mechanisms involved in BP induced cell death in human prostate cancer cells.

Methods/Principal Findings

Two human prostate cancer cell lines, PC-3 and LNCaP, were treated with BP, and subsequently evaluated for their viability and cell cycle profiles. BP caused cell cycle arrest and cell death in both cell lines. The G0/G1 phase arrest was correlated with increase levels of CDK inhibitors (p16, p21 and p27) and decrease of the checkpoint proteins. To determine the mechanisms of BP-induced growth arrest and cell death in prostate cancer cell lines, we performed a microarray study to identify alterations in gene expression induced by BP in the LNCaP cells. Several BP-induced genes, including the GADD153/CHOP, an endoplasmic reticulum stress (ER stress)-regulated gene, were identified. BP-induced ER stress was evidenced by increased expression of the downstream molecules GRP78/BiP, IRE1-α and GADD153/CHOP in both cell lines. Blockage of IRE1-α or GADD153/CHOP expression by siRNA significantly reduced BP-induced cell death in LNCaP cells. Furthermore, blockage of JNK1/2 signaling by JNK siRNA resulted in decreased expression of IRE1-α and GADD153/CHOP genes, implicating that BP-induced ER stress may be elicited via JNK1/2 signaling in prostate cancer cells. BP also suppressed LNCaP xenograft tumor growth in NOD-SCID mice. It caused 68% reduction in tumor volume after 18 days of treatment.

Conclusions

Our results suggest that BP can cause G0/G1 phase arrest in prostate cancer cells and its cytotoxicity is mediated by ER stress induction. Thus, BP may serve as an anticancer agent by inducing ER stress in prostate cancer.     

Effects of exposure of different skin areas to low-power laser light

The effect of He-Ne laser light of extremely low power (632.8 nm, 0.2 mW/cm²) on the immune status of mice bearing solid tumors was studied. The state of tumor-bearing animals was assessed taking into account the number of immunocompetent cells, concentration of cytokines (tumor necrosis factor and interleukin-2), production of nitric oxide, expression of heat shock proteins 70 and 90, and the activity of natural killer cells. The model of a solid tumor was formed by subcutaneous transplantation of Ehrlich ascite carcinoma cells to mice; the average lifespan of animals was approximately 55 days. Different areas of skin of tumor-bearing mice were irradiated with laser light either singly (1 min; dose, 0.012 J/cm²) or repeatedly (1 min every 3 days over 30 days; total dose, 0.1 J/cm²). It was established that long-term chronic exposure of mice bearing Ehrlich ascite carcinoma cells to low-power laser light in the thymus projection area and especially in the tumor projection area leads to a decrease in the natural antitumor potential, which is manifested in acceleration of tumor growth and a tendency to decrease in the lifespan of tumor-bearing mice. Conversely, stimulation of antitumor immunity was observed over several days after a single exposure to low-power laser radiation. The results suggest that it is expedient to continue studies of the immunomodulating effects of low-power laser light and demonstrate the necessity of monitoring the immune system in the course of laser therapy.     

Tests of mutagenesis and reproduction in male rats exposed to 2,450-MHz (CW) microwaves

Tests of mutagenesis and reproduction were conducted in male rats which were irradiated by 2,450-MHz, continuous-wave (CW) microwaves, 4 hr/day from day 6 of gestation to 90 days of age at 5 mW/cm²; or 5 hr/day for five days beginning on the 90th day of age at 10 mW/cm²; or 4 hr/day, 5 days/ wk for four weeks, beginning on the 90th day of age. During selected weekly periods after treatment, the rats were bred to pairs of untreated, normal female rats that were examined in late pregnancy by means of the dominant lethal assay. The reproductive efficiency of these males, as reflected in their breeding, was also examined for changes relating to their microwave experience. No significant evidence of germ-cell mutagenesis was detected when data of microwave-exposed males were compared with those of sham-exposed males, even though there were significant increases in rectal and intra-testicular temperatures at a power density of 28 mW/cm². Temporary sterility, as indexed by fewer pregnancies, was seen at the highest power density.     

Effects of Pulsed Microwave Radiation Pre -and Post-Natally on the Developing Brain in Mice

Pregnant mice were irradiated for 5 hours daily throughout the pregnancy with pulsed microwaves at an incident average power density of 8 mW/cm², or were sham irradiated. After birth, from day 3 to day 20, half the offspring delivered by irradiated mice were irradiated (RR group) and half were sham-irradiated (RC group). The same procedure was used for offspring delivered by sham-irradiated mice (CR and CC). All offspring were sacrificed at 22 days of age. Histochemical analyses of the hypothalamus and liver were performed with a microspectrophotometer. The data suggested that succinate dehydrogenase in the hypothalamus was reduced by either pre-or post-natal microwave exposure. Similar changes occurred in the liver. The same pattern appeared with hypothalamic catechol-amine and monoamine oxidase. The data indicate that low-intensity microwave exposure can induce subtle alterations in offspring not detected with previously used techniques.     

Regenerative Activity of Muscle Allografts and the State of Thymus under Conditions of Long-Term Laser Treatment of Adrenal Glands and Transplantation Site Prior to Surgery

Both gastrocnemius muscles were cross-allografted between the intact rats and the rats whose adrenal glands and right shins were exposed to He–Ne laser radiation before the surgery (632.8 nm, 2.5–3.0 mW/cm², 10 sessions over 14 days at a cumulative dose of 15–18 J/cm² per rat). By the time of grafting, the activity of adrenal glands decreased, while the activity of thymus increased. As a result, the allogenic muscle tissue was resorbed more actively in the irradiated rats, especially in the right, irradiated shin. At the same time, reinnervation of the grafts containing a certain amount of survived allogenic muscle tissue was more active in the right shin, compared to the left, unirradiated one. Survival of the donor muscle tissue and its improved reinnervation were observed in a higher proportion of grafts in the intact rats. The allografts from the right (preirradiated) shin of animals with irradiated adrenal glands proved to be more viable, compared to the left (unirradiated) shin.     

Effects of 2.45-GHz microwave radiation and phorbol ester 12-O-tetradecanoylphorbol-13-acetate on dimethylhydrazine-induced colon cancer in mice

The purpose of this study was to investigate the effects of 2.45 GHz microwave (MW) radiation on dimethylhydrazine (DMH)-induced colon cancer in mice. The subjects were 115 Balb/c mice 4 weeks of age. The animals were divided into group A (control), group B (DMH), group C (DMH + MW), and group D [DMH + 12-O-tetradecanoylphorbol-13-acetate (TPA)]. Radiation (10 mW/cm²) was delivered dorsally with the E field parallel to the mouse's long body axis in an anechoic chamber. Radiations were administered 3 hr daily, 6 days per week, over a period of 5 months. The average SAR was estimated to be 10–12 W/kg. During the course of radiation treatments, DMH was injected once per week. The tumor promoter TPA was administered once per week for 10 weeks, from the third week on, after the initial treatment. The incidence of tumors did not significantly differ between the three test groups (groups B, C, and D; P > 0.25). However, the number of tumors, the size of the tumors, and the incidence of protuberant and infiltrative types in tumor-bearing animals were higher in group D compared to groups B and C (P < 0.05). No difference was found between groups B and C (P > 0.25). The study indicates that 2.45 GHz microwave radiation at 10 mW/cm² power density did not promote DMH-induced colon cancers in young mice. The study also showed that TPA could accelerate colon tumor production if a tumor was initiated. © 1994 Wiley-Liss, Inc.     

Microwaves modify thermoregulatory behavior in squirrel monkey

Squirrel monkeys (Saimiri sciureus) trained to regulate environmental temperature (T_a) behaviorally were exposed in the far field of a horn antenna to ten-minute periods of 2,450 MH_z CW microwaves. Incident power density ranged from 1 to 22 mW/cm². The corresponding specific absorption rate (SAR), derived from temperature increments in saline-filled styrofoam models, ranged from 0.15 to 3.25 W/kg. Controls included exposure to infrared radiation of equivalent incident energy and no radiation exposure. Normal thermoregulatory behavior produces tight control over environmental and body temperatures; most monkeys select a T_a of 34–36°C. Ten-minute exposures to 2,450 MH_z CW microwaves at an incident power density of 6–8 mW/cm² stimulated all animals to select a lower T_a. This threshold energy represents a whole-body SAR of 1.1 W/kg, about 20% of the resting metabolic rate of the monkey. Thermoregulatory behavior was highly efficient, and skin and rectal temperatures remained stable, even at 22 mW/cm² where the preferred T_a was lowered by as much as 4°C. No comparable reduction in selected T_a below control levels occurred during exposure to infrared radiation of equal incident power density.     

Microwave radiation (2450 MHz) alters the endotoxin-induced hypothermic response of rats

The parenteral administration of bacterial endotoxin to rats causes a hypothermia that is maximal after approximately 90 minutes. When endotoxin-injected rats were held in a controlled environment at 22°C and 50% relative humidity and exposed for 90 minutes to microwaves (2450 MHz, CW) at 1 mW/cm², significant increases were observed in body temperature compared with endotoxintreated, sham-irradiated rats. The magnitude of the response was related to power density (10 mW/cm² > 5 mW/cm² > 1 mW/cm²). Saline-injected rats exposed for 90 minutes at 5 mW/cm² (specific absorption rate approximately 1.0 mW/g) showed no significant increase in body temperature compared with saline-injected, sham-irradiated rats. The hypothermia induced by endotoxin in rats was also found to be affected by ambient temperature alone. Increases in ambient temperature above 22°C in the absence of microwaves caused a concomitant increase in body temperature. This study reveals that subtle microwave heating is detectable in endotoxin-treated rats that have an impaired thermoregulatory capability. These results indicate that the interpretation of microwave-induced biological effects observed in animals at comparable rates and levels of energy absorption should include a consideration of the thermogenic potential of microwaves.     

Cytokine profiles in mice tissues after irradiation of the thymus projection area with femtosecond laser

The effects of pulsed femtosecond laser irradiation in the near ultraviolet region on the levels of cytokines in the thymus, blood, and skin of irradiated mice have been studied. Irradiation of the thymus projection area with low-intensity laser radiation in the near UV region of the spectrum showed significant changes in cytokine levels in the skin and thymus and, to a lesser extent, in the blood of irradiated mice. Laser irradiation with a power density of 20 mW/cm² affects the cytokine profile in the thymus: IFN-γ, IL-3, IL-4, IL-5, eotaxin, GM-CSF, and chemokine KC–factors that can affect differentiation and proliferation of the cells of the immune system in the gland. Conclusions: It is assumed that changes in the expression of cytokines in the thymus after laser irradiation are explained by the rearrangement of biochemical processes possibly associated with the maturation of cells in the gland.     

Humoral and cell-mediated immune function in adult japanese quail following exposure to 2.45-GHz microwave radiation during embryogeny

Japanese quail, Coturnix coturnix japonica, eggs were subjected to 2.45-GHz CW microwave radiation at 5 mW/cm² (SAR = 4.03 mW/g) during the first 12 days of embryogeny. Following hatching the exposed embryos, as well as nonexposed controls, were reared to 22 weeks of age. Humoral immune potential, as indicated by comparable anti-CRBC antibody, IgM and IgG, levels at 0, 4, and 7 days postimmunization in both exposed and control quail was not affected significantly. However, cell-mediated immune potential, measured by the reaction to intradermal injection of phytohemagglutinin-P in the wing web, was reduced in the exposed females, but not in the exposed males. Additionally, total leukocyte numbers and absolute circulating numbers of lymphocytes, monocytes, and heterophils were increased significantly only in the exposed females. These data show that exposure of Japanese quail during embryogenesis reduced cell-mediated immune potential and induced a general leukocytosis in females.     

Effect of intermittency factor on singlet oxygen and PGE2 formation in azulene-mediated photodynamic therapy: A preliminary study

In photodynamic therapy, intermittent irradiation modes that incorporate an interval between pulses are believed to decrease the effect of hypoxia by permitting an interval of re-oxygenation. The effect of the irradiation intermittency factor (the ratio of the irradiation pulse time to the total irradiation time) on singlet oxygen formation and inflammatory cytokine production was examined using azulene as a photosensitizer. Effects of difference intermittency factor on singlet oxygen formation and inflammatory cytokine were examined. Azulene solutions (1/10 μM) were irradiated with a 638-nm 500 mW diode laser in fractionation (intermittency factor of 5 or 9) or continuous mode using 50 mW/cm² at 4 or 8 J/cm². Singlet oxygen measurement was performed using a dimethyl anthracene probe. Peripheral blood mononuclear cells (PBMC) were stimulated by 10 ng/ml rhTNF-α for 6 h, before addition of 1 and 10 μM azulene solutions and irradiation. PGE₂ measurement was undertaken using a human PGE₂ ELISA kit. Kruskal-Wallis with Dunn Bonferroni test was used for statistical analyses at p < 0.05.Irradiation of 1 μM azulene+4 J/cm²+intermittency factor of 9 increased singlet oxygen 3-fold (p < 0.0001). Irradiation of 10 μM azulene at either 4 J/cm²+intermittency of 9 or 8 J/cm²+intermittency factor of 5 reduced PGE₂ expression in PBMCs to non-inflamed levels. Thus, at 50 mW/cm², 10 μM azulene-mediated photodynamic therapy with a high intermittency factor and a low energy density generated sufficient singlet oxygen to suppress PGE₂ in Inflamed PBMCs.