The effect of anoxia on fowl spermatozoa: Recovery of fertilizing ability,motility, and cellular concentrations of potassium and ATP

Fowl semen when diluted in a glutamate-based medium without glucose, gradually lost its fertilizing ability during 4 hr of anaerobic incubation at 30°C. This incubation regime offered a system by which various in-vitro tests of spermatozoal viability could be assessed for their usefulness as monitors of fertilizing ability. Widely used tests such as spermatozoal enzyme leakage, dye exclusion, and morphology as assessed by light microscopy showed no change in spermatozoal status as the fertilizing ability declined. However the ability of sperm, during a short aerobic incubation to restore their motility and ATP and K⁺ concentrations, declined as did their fertilizing ability. When glucose was added to this re-aeration medium, spermatozoal motility, K⁺ and ATP concentrations, and fertilizing ability were restored to optimal levels. Thus the fertilizing ability of fowl sperm, following anaerobic storage at 30°C, appeared to be related to their ability to restore ATP and K⁺ concentrations and motility. An initial event in the loss of fertilizing ability was a loss in the ability of sperm to oxidise endogenous substrates. This could be restored by the addition of glucose.     

Behavioural and autonomic thermoregulation in hamsters during microwave-induced heat exposure

1. 1.|Preferred ambient temperature (T_a) and ventilatory frequency were measured in free-moving hamsters exposed to 2450 MHz microwaves. A waveguide exposure system which permits continuous monitoring of the absorbed heat load accrued from microwave exposure was imposed with a longitudinal temperature gradient which allowed hamsters to select their preferred T_a. Ventillatory frequency was monitored remotely by analysing the rhythmic shifts in unabsorbed microwave energy passing down the waveguide.

2. 2.|Without microwave exposure hamsters selected an average T_a of 30.2°C. This preferred T_a did not change until the rate of heat absorption (SAR) from microwave exposure exceeded approx. 2 W kg⁻¹. In a separate experiment, a SAR of 2.0 W kg⁻¹ at a T_a of 30°C was shown to promote an average 0.5°C increase in colonic temperature. Hamsters maintained their ventilatory frequency at baseline levels by selecting a cooler T_a during microwave exposure. In contrast, hamsters maintained at a T_a of 30°C (without a temperature gradient) underwent a sharp increase in ventilatory frequency compared to animals allowed to select their own T_a.

3. 3.|These data support previous studies suggesting that during thermal stress behavioural thermoregulation (i.e. preferred T_a) takes prescedence over autonomic thermoregulation (i.e. ventilatory frequency). It is apparent that selecting a cooler T_a is a more efficient and/or effective than autonomic thermoregulation for dissipating a heat load accrued from microwave exposure.

Operant control of convective cooling and microwave irradiation by the squirrel monkey

Adult male squirrel monkeys (Saimiri sciureus) were individually chair-restrained in an air-conditioned Styrofoam box in the far field of a horn antenna. Each monkey first received extensive training to regulate the temperature of the air circulating through the box by selecting between 10 and 50 degrees C air source temperatures. Then, to investigate the ability of the animals to utilize microwaves as a source of thermalizing energy, 2450-MHz continuous wave microwaves accompanied by thermoneutral (30 degrees C) air were substituted for the 50 degrees C air. Irradiation at each of three power densities was made available, ie, at 20, 25, and 30 mW/cm2 [SAR = 0.15 (W/kg)/(mW/cm2)]. The percentage of time that the monkeys selected microwave irradiation paired with thermoneutral air averaged 90% at 20 and at 25 mW/cm2. The mean percentage declined reliably (p less than 0.001) to 81% at 30 mW/cm2, confirming the monkey's ability to utilize microwave irradiation as a source of thermal energy during the course of behavioral thermoregulation. All animals readily made the warm-air to microwave-field transition, regulating rectal temperature with precision by sequentially selecting 10 degrees C air, then microwave irradiation accompanied by 30 degrees C air. Although the selection of cooler air resulted in a slight reduction of skin temperatures, normal rectal temperature was maintained. The results indicate that the squirrel monkey can utilize a microwave source in conjunction with convective cooling to regulate body temperature behaviorally.     

Comparative effects of extremely high power microwave pulses and a brief CW irradiation on pacemaker function in isolated frog heart slices

The existence of specific bioeffects due to high peak power microwaves and their potential health hazards are among the most debated but least explored problems in microwave biology. The present study attempted to reveal such effects by comparing the bioeffects of short trains of extremely high power microwave pulses (EHPP, 1 micros width, 250-350 kW/g, 9.2 GHz) with those of relatively low power pulses (LPP, 0.5-10 s width, 3-30 W/g, 9.2 GHz). EHPP train duration and average power were made equal to those of an LPP; therefore both exposure modalities produced the same temperature rise. Bioeffects were studied in isolated, spontaneously beating slices of the frog heart. In most cases, a single EHPP train or LPP immediately decreased the inter-beat interval (IBI). The effect was proportional to microwave heating, fully reversible, and easily reproducible. The magnitude and time course of EHPP- and LPP-induced changes always were the same. No delayed or irreversible effects of irradiation were observed. The same effect could be repeated in a single preparation numerous times with no signs of adaptation, sensitization, lasting functional alteration, or damage. A qualitatively different effect, namely, a temporary arrest of preparation beats, could be observed when microwave heating exceeded physiologically tolerable limits. This effect also did not depend on whether the critical temperature rise was produced by LPP or EHPP exposure. Within the studied limits, we found no indications of EHPP-specific bioeffects. EHPP- and LPP-induced changes in the pacemaker rhythm of isolated frog heart preparation were identical and could be entirely attributed to microwave heating.     

微波辐射对桃蚜生长发育和繁殖的影响

【目的】研究筛选对桃蚜Myzus persicae有致死作用的安全微波频率和照射时长,以为探究新型物理防蚜技术,弥补化学防治上的缺陷提供参考依据。【方法】在暗箱中,应用微波发射仪分别发射1375, 2 750, 5 500和11 000 MHz 4个不同频率的微波照射桃蚜1日龄无翅成蚜,每个频率的照射时长分别为15, 30, 60和120 s;照射后在人工气候箱中饲养,分别于照射后8, 24, 48和72 h观察其生长发育及繁殖状况,统计桃蚜死亡率、繁殖力(累计产蚜量)及子代有翅蚜率。【结果】4个不同频率的微波分别在4个不同照射时长下,对桃蚜1日龄无翅成蚜的死亡率、繁殖力和子代翅型分化都有不同程度的影响。照射后72 h, 5 500 MHz微波照射时间为15 s时对桃蚜1日龄无翅成蚜的致死作用最强,死亡率达到55.00%,在照射时间为30和120 s时可抑制子代桃蚜向有翅蚜的分化。2 750 MHz微波照射30和60 s时促进桃蚜1日龄成蚜繁殖,照射30 s时繁殖力最强,而照射15和120 s时却表现为抑制繁殖,且2 750 MHz微波照射30 s能抑制子代桃蚜向有翅蚜分化。【结论】微波辐射能够影响桃蚜1日龄成蚜的存活、繁殖和子代翅型分化。本研究初步筛选出了对桃蚜1日龄无翅成蚜有致死作用的微波频率和照射时长。     

Safranin O Staining Using a Microwave Oven

We investigated the effects of microwave irradiation on a safranin O staining method for paraffin sections of formalin fixed rabbit larynx. The control sections were stained according to the conventional method, and the experimental sections were stained in microwave oven for 10 sec at 360 W in Weigert's iron hematoxylin, and for 30 sec at 360 W in fast green and 0.1% safranin O staining solutions. Light microscopic examination of the sections revealed that the microwave heating did not adversely affect the staining properties of cartilage tissue compared to the conventional staining method. Small differences such as darker staining of the matrix and shrinkage of the cytoplasm was observed in some microwave treated sections. The present study revealed that microwave application can be used safely for the safranin O method with the advantage of reduced staining time.     

Safranin O Staining Using a Microwave Oven

We investigated the effects of microwave irradiation on a safranin O staining method for paraffin sections of formalin fixed rabbit larynx. The control sections were stained according to the conventional method, and the experimental sections were stained in microwave oven for 10 sec at 360 W in Weigert's iron hematoxylin, and for 30 sec at 360 W in fast green and 0.1% safranin O staining solutions. Light microscopic examination of the sections revealed that the microwave heating did not adversely affect the staining properties of cartilage tissue compared to the conventional staining method. Small differences such as darker staining of the matrix and shrinkage of the cytoplasm was observed in some microwave treated sections. The present study revealed that microwave application can be used safely for the safranin O method with the advantage of reduced staining time.     

Thermoregulatory responses of the immature rat following repeated postnatal exposures to 2,450-MHz microwaves

This study was designed to determine the changes that occur in the thermoregulatory ability of the immature rat repeatedly exposed to low-level microwave radiation. Beginning at 6-7 days of age, previously untreated rats were exposed to 2,450-MHz continuous-wave microwaves at a power density of 5 mW/cm2 for 10 days (4 h/day). Microwave and sham (control) exposures were conducted at ambient temperatures (Ta) which represent different levels of cold stress for the immature rat (ie, "exposure" Ta = 20 and 30 degrees C). Physiological tests were conducted at 5-6 and 16-17 days of age, in the absence of microwaves, to determine pre- and postexposure responses, respectively. Measurements of metabolic rate, colonic temperature, and tail skin temperature were made at "test" Ta = 25.0, 30.0, 32.5, and 35.0 degrees C. Mean growth rates were lower for rats exposed to Ta = 20 degrees C than for those exposed to Ta = 30 degrees C, but microwave exposure exerted no effect at either exposure Ta. Metabolic rates and body temperatures of all exposure groups were similar to values for untreated animals at test Ta of 32.5 degrees C and 35.0 degrees C. Colonic temperatures of rats repeatedly exposed to sham or microwave conditions at exposure Ta = 20 degrees C or to sham conditions at exposure Ta = 30 degrees C were approximately 1 degrees C below the level for untreated animals at test Ta of 25.0 degrees C and 30.0 degrees C. However, when the exposure Ta was warmer, rats exhibited a higher colonic temperature at these cold test Ta, indicating that the effectiveness of low-level microwave treatment to alter thermoregulatory responses depends on the magnitude of the cold stress.     

Increased serum enzyme activity in microwave-exposed rats

Heat stable serum enzymes were studied in rats exposed to microwaves (2.45 GHz, 120 Hz amplitude modulated) 24 hr after a single 4-hr exposure or immediately after 3 and 10 exposures to 0.1 to 55 mW/cm2. In addition, stable colonic temperature at 41.5 degrees C for 30 min was maintained by microwave exposure in a group of five rats under barbiturate anesthesia. Alkaline phosphatase and lactic dehydrogenase did not increase as a result of microwave exposure. Increased serum glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminase (GOT) were noted in the 41.5 degrees C group 24 hr after exposure. A threshold body temperature for acute cellular injury after microwave exposure was demonstrated. The acute cellular injury could be in the liver. These mild elevations in the serum enzyme levels (mean +/- SE, GOT = 167 +/- 40 U/liter: GPT = 74 +/- 26 U/liter) indicated that the injuries were not accompanied by any significant sequelae in the rat. From this threshold and colonic temperature (41.5 degrees C for 30 min) in barbiturate-anesthetized, microwave-exposed rats, we derived a tentative threshold for the whole-body average absorption rate at 14 W/kg (70 mW/cm2 at 2.45 GHz for adult rats) for 4 hr. This tentative threshold is subject to changes by duration of exposure and by compounding variables influencing maintenance of body temperature.     

Absence of a synergistic effect between moderate-power radio-frequency electromagnetic radiation and adriamycin on cell-cycle progression and sister-chromatid exchange.

In our laboratories we are conducting investigations of potential interactions between radio-frequency electromagnetic radiation (RFR) and chemicals that are toxic by different mechanisms to mammalian cells. The RFR is being tested at frequencies in the microwave range and at different power levels. We report here on the 1) ability of simultaneous RFR exposures to alter the distribution of cells in first and second mitoses from that after treatment by adriamycin alone, and 2) on the ability of simultaneous RFR exposure to alter the extent of sister chromatid exchanges (SCEs) induced by adriamycin alone. This chemical was selected because of its reported mechanism of action and because it is of interest in the treatment of cancer. In our studies, Chinese hamster ovary (CHO) cells were exposed for 2 h simultaneously to adriamycin and pulsed RFR at a frequency of 2,450 MHz and a specific absorption rate of 33.8 W/Kg. The maximal temperature (in the tissue-culture medium) was 39.7 +/- 0.2 degrees C. The experiments were controlled for chemical and RFR exposures, as well as for temperature. Verified statistically, the data indicate that the RFR did not affect changes in cell progression caused by adriamycin, and the RFR did not change the number of SCEs that were induced by the adriamycin, which adriamycin is known to affect cells by damaging their membranes and DNA.     

In vitro cytogenetic effects of 2450 MHz waves on human peripheral blood lymphocytes

Cytogenetic analyses were performed on human peripheral blood lymphocytes exposed to 2450 MHz microwaves during 30 and 120 min at a constant temperature of 36.1°C (body temperature). The temperature was kept constant by means of a temperature probe put in the blood sample which gives feedback to a microcomputer that controls the microwave supply. We found a marked increase in the frequency of chromosome aberrations (including dicentric chromosomes and acentric fragments) and micronuclei. On the other hand the microwave exposure did not influence the cell kinetics nor the sister chromatid exchange (SCE) frequency. ©1993 Wiley-Liss, Inc.     

Calreticulin attenuated microwave radiation-induced human microvascular endothelial cell injury through promoting actin acetylation and polymerization

Recent work reveals that actin acetylation modification has been linked to different normal and disease processes and the effects associated with metabolic and environmental stressors. Herein, we highlight the effects of calreticulin on actin acetylation and cell injury induced by microwave radiation in human microvascular endothelial cell (HMEC). HMEC injury was induced by high-power microwave of different power density (10, 30, 60, 100 mW/cm², for 6 min) with or without exogenous recombinant calreticulin. The cell injury was assessed by lactate dehydrogenase (LDH) activity and Cell Counting Kit-8 in culture medium, migration ability, intercellular junction, and cytoskeleton staining in HMEC. Western blotting analysis was used to detected calreticulin expression in cytosol and nucleus and acetylation of globular actin (G-actin). We found that HMEC injury was induced by microwave radiation in a dose-dependent manner. Pretreatment HMEC with calreticulin suppressed microwave radiation-induced LDH leakage and increased cell viability and improved microwave radiation-induced decrease in migration, intercellular junction, and cytoskeleton. Meanwhile, pretreatment HMEC with exogenous calreticulin upregulated the histone acetyltransferase activity and the acetylation level of G-actin and increased the fibrous actin (F-actin)/G-actin ratio. We conclude that exogenous calreticulin protects HMEC against microwave radiation-induced injury through promoting actin acetylation and polymerization.     

Temporal bisection in rats: The effects of high-peak-power pulsed microwave irradiation

The effects of high-peak-power, pulsed microwaves on a time perception and discrimination task were studied in rats. Exposures were performed with the TEMPO exposure system, which produces an 80 nanosecond pulse with peak-power levels in excess of 700 megawatts. The ability to expose animals to such fields within a controlled environment is unique. As determined by calorimetry, a maximal, whole-body-averaged, specificabsorption rate of 0.072 W/kg was produced. Thus exposures were well below a recommended SAR limit of 0.4 W/kg. Power levels of transmitted microwaves were varied over a 50 dB range to obtain ascending and descending dose-response functions for each of the behavioral measures. Measures of time perception, response bias, and total trials did not change with power level. Dose-response effects were observed for discriminability (ability to distinguish between durations), session time, and trial completions (null responses, failures to respond on a trial). Covarying sound and X-ray exposures produced by TEMPO did not reliably correlate with the observed microwave effects. The observation of repeatable dose-response effects on discriminability and null responses indicates that the microwave exposures were affecting cognitive function in the rats, particularly the decision-making process. © 1993 wiley-Liss, Inc.     

New Developed Cylindrical TM010 Mode EPR Cavity for X-Band In Vivo Tooth Dosimetry

EPR tooth in vivo dosimetry is an attractive approach for initial triage after unexpected nuclear events. An X-band cylindrical TM010 mode resonant cavity was developed for in vivo tooth dosimetry and used in EPR applications for the first time. The cavity had a trapezoidal measuring aperture at the exact position of the cavity’s cylindrical wall where strong microwave magnetic field H1 concentrated and weak microwave electric field E1 distributed. Theoretical calculations and simulations were used to design and optimize the cavity parameters. The cavity features were evaluated by measuring DPPH sample, intact incisor samples embed in a gum model and the rhesus monkey teeth. The results showed that the cavity worked at designed frequency and had the ability to make EPR spectroscopy in relative high sensitivity. Sufficient modulation amplitude and microwave power could be applied into the aperture. Radiation induced EPR signal could be observed remarkably from 1 Gy irradiated intact incisor within only 30 seconds, which was among the best in scan time and detection limit. The in vivo spectroscopy was also realized by acquiring the radiation induced EPR signal from teeth of rhesus monkey whose teeth was irradiated by dose of 2 Gy. The results suggested that the cavity was sensitive to meet the demand to assess doses of significant level in short time. This cavity provided a very potential option for the development of X-band in vivo dosimetry.     

Oil extraction from Scenedesmus obliquus using a continuous microwave system--design, optimization, and quality characterization

A 1.2 kW, 2450 MHz resonant continuous microwave processing system was designed and optimized for oil extraction from green algae (Scenedesmus obliquus). Algae-water suspension (1:1 w/w) was heated to 80 and 95°C, and subjected to extraction for up to 30 min. Maximum oil yield was achieved at 95°C and 30 min. The microwave system extracted 76-77% of total recoverable oil at 20-30 min and 95°C, compared to only 43-47% for water bath control. Extraction time and temperature had significant influence (p<0.0001) on extraction yield. Oil analysis indicated that microwaves extracted oil containing higher percentages of unsaturated and essential fatty acids (indicating higher quality). This study validates for the first time the efficiency of a continuous microwave system for extraction of lipids from algae. Higher oil yields, faster extraction rates and superior oil quality demonstrate this system's feasibility for oil extraction from a variety of feedstock.     

Lethal Temperature for Pinewood Nematode, Bursaphelenchus xylophilus, in Infested Wood Using Microwave Energy

To reduce the risks associated with global transport of wood infested with pinewood nematode Bursaphelenchus xylophilus, microwave irradiation was tested at 14 temperatures in replicated wood samples to determine the temperature that would kill 99.9968% of nematodes in a sample of ≥ 100,000 organisms, meeting a level of efficacy of Probit 9. Treatment of these heavily infested wood samples (mean of > 1,000 nematodes/g of sapwood) produced 100% mortality at 56 °C and above, held for 1 min. Because this "brute force" approach to Probit 9 treats individual nematodes as the observational unit regardless of the number of wood samples it takes to treat this number of organisms, we also used a modeling approach. The best fit was to a Probit function, which estimated lethal temperature at 62.2 (95% confidence interval 59.0-70.0) °C. This discrepancy between the observed and predicted temperature to achieve Probit 9 efficacy may have been the result of an inherently limited sample size when predicting the true mean from the total population. The rate of temperature increase in the small wood samples (rise time) did not affect final nematode mortality at 56 °C. In addition, microwave treatment of industrial size, infested wood blocks killed 100% of > 200,000 nematodes at ≥ 56 °C held for 1 min in replicated wood samples. The 3(rd)-stage juvenile (J3) of the nematode, that is resistant to cold temperatures and desiccation, was abundant in our wood samples and did not show any resistance to microwave treatment. Regression analysis of internal wood temperatures as a function of surface temperature produced a regression equation that could be used with a relatively high degree of accuracy to predict internal wood temperatures, under the conditions of this study. These results provide strong evidence of the ability of microwave treatment to successfully eradicate B. xylophilus in infested wood at or above 56 °C held for 1 min.     

Dominant lethal studies in male mice after exposure to 2.45 GHz microwave radiation

Adult male mice had the lower halves of their bodies exposed in a waveguide system to 2.45 GHz microwave radiation for 30 min. The half body dose-rate of 43 W kg-1 had been shown in a previous study [7] to deplete severely the heat-sensitive stages of sperm production. The males were mated at intervals to adult hybrid females over the following 8-10 weeks. There was no significant reduction in post-implantation survival, suggesting that the microwave exposure did not have a mutagenic effect on the male germ cells. However, pregnancy rate was significantly reduced in weeks 3, 4, 5 and 6; reaching a minimum of about 10% of the control value in weeks 4 and 5. The occurrence of low values in weeks 4 and 5 correlated well with the expected reductions in sperm count due to the pattern of depletion of the spermatogenic epithelium of the testes. Thus it was concluded that the reduced pregnancy rate resulted from reduced male fertility. Pre-implantation survival can also be affected by reduced sperm count [8] and was significantly reduced in this study but it correlated less well with the anticipated heat response. A further study is in progress looking at the contribution of sperm count and sperm abnormality to the results.     

The effect of modification of cytosines in Escherichia coli 16-S rRNA on reconstitution and function of 30-S ribosomes.

O-Methylhydroxylamine (methoxyamine) was used for selective modification of cytosine residues in Escherichia coli 16-S rRNA. It was shown that cytosines accessible for methoxyamination are randomly distributed along the 16-S rRNA chain. Preparations of methoxyaminated 16-S rRNA, containing 2--130 modified cytosines/chain, still retained the ability to bind 30-S proteins, but the physical assembly of reconstituted particles was incorrect. The protein compositions of the reconstituted and native particles did not differ qualitatively from each other. However, the amount of protein in reconstituted particles decreased with an increasing number of methoxyaminated cytosines in 16-S rRNA. The particles obtained sedimented slower than native 30-S subunits, lost their ability to associate with 50-S ribosomes and to bind native phage f2 RNA. In contrast, modification of 16-S rRNA did not affect binding of poly(U) by reconstituted particles.     

Stable and efficient immobilization technique of aldolase under consecutive microwave irradiation at low temperature

To establish a stable and efficient immobilization technique under microwave irradiation, a focused microwave reaction system was used, where the temperature was set appropriately in the microwave system and cooling module to produce consecutive microwave irradiation. 2-Deoxy-D-ribose-5-phosphate aldolase (DERA) was rapidly and efficiently immobilized in mesocellular siliceous foams (MCFs) under microwave irradiation. When the output power in the microwave system was set to 30 W, after 3 min, 88.4% of the enzyme protein was coupled to the wall of the support pores and the specific activity of the immobilized enzyme was 2.24 U mg⁻¹, 149.2% higher than that of the free enzyme and 157.0% higher than that of the non-microwave-assisted immobilized enzyme. In catalysis, microwave-assisted immobilized DERA tolerated a wider range of both pH and temperature than other DERA preparations. The thermal and storage stabilities were also significantly improved. This focused; microwave-assisted immobilization technique has proven to be simple, stable and highly efficient. This technique could also be applied to other enzyme immobilizations.     

Leukocyte numbers in hemorrhaged Japanese quail after microwave irradiation in ovo

1. Coturnix coturnix japonica eggs were exposed to 2.45-GHz continuous wave microwave radiation at an incident power density of 5 mW/cm2 (and a specific rate of 4 mW/g) during the first 12 days of embryogeny. After hatching, hematological changes in response to an acute hemorrhage were measured in exposed and nonexposed (control) juveniles of both sexes. 2. Exposure did not affect erythroid cell numbers either before or after hemorrhage. 3. Exposure affected the recovery of lymphocyte and heterophil numbers after hemorrhage, but the effect was sex-limited. 4. These data indicate that microwave irradiation during embryogeny in ovo affects the ability of Japanese quail to recover from an acute and voluminous hemorrhage and that these radiation effects are sex-limited and consistent with a previous report.