The effect of collagen degradation products (CDP) on the central dopaminergic system

The effect of collagen degradation products (CDP I--molecular weight circa 3000 D and CDP II--molecular weight circa 1200 D) on the central dopaminergic system was studied. Differences in the action of both these fractions in the apomorphine stereotypy test were noted; CDP I administered to rats in a dose of 5 micrograms just before application of the drug inducing the stereotypy enhanced the stereotypic behaviour of the animals whereas a dose of 40 micrograms significantly inhibited such behaviour. CDP II, on the other hand, had no effect on this type of stereotypy. Both fractions given in doses of 15 and 40 micrograms enhanced the stereotypy induced by amphetamine. CDP I and CDP II (15 and 40 micrograms) administered 30 min before observation of the animals intensified the cataleptic action of haloperidol, whereas both fractions (CDP I and CDP II) when administered 45 min before observation reduced the catalepsy.     

Research on the neurological effects of nonionizing radiation at the University of Washington.

This paper reviews research on neurological effects of low-level microwave irradiation, which was performed at the University of Washington, during the decade of the 1980s. We studied in the rat the effects of microwave exposure on the actions of various psychoactive drugs, on the activity of cholinergic systems in the brain, and on the neural mechanisms involved. Our results indicate that endogenous opioids play an important mediating role in some of the neurological effects of microwaves, and that parameters of microwave exposure are important determinants of the outcome of the microwave effects.     

Microwave facilitation of domperidone antagonism of apomorphine-induced stereotypic climbing in mice

The dopaminergic agonist apomorphine produced dose-dependent stereotypic climbing behavior in mice housed in cages with vertical bars. This drug effect was competitively inhibited by systemic pretreatment with the centrally acting dopaminergic antagonist haloperidol but not by microwave irradiation (2.45 GHz, 20 mW/cm2, CW, 10 min) nor by systemic pretreatment with domperidone, a dopaminergic antagonist that only poorly penetrates the blood-brain barrier (BBB). Yet when mice were systemically pretreated with domperidone and then subjected to microwave irradiation (as above), the apomorphine effect was significantly reduced. Microwave irradiation also facilitated antagonism of the apomorphine effect by low and otherwise ineffective systemic pretreatment doses of haloperidol. Apomorphine-induced stereotypic climbing behavior was also reduced by domperidone administered intracerebrally, which bypassed the BBB. Exposure of intracerebral domperidone-pretreated animals to microwave irradiation failed to increase the degree of antagonism. These findings indicate that microwave irradiation can facilitate central effects of domperidone, a drug which acts mainly in the periphery. One possible explanation for these findings is that microwave irradiation alters the permeability of the BBB and increases the entry of domperidone to central sites of action.     

Effect of Microwave and He-Ne Laser on Enzyme Activity and Biophoton Emission of Isatis indigotica Fort

The seed embryos of Isatis indigotica Fort were exposed to He-Ne laser (5.23 mW/mm2, radiated for 5 min) and microwave (1.26 mW/mm2, radiated for 8 s) irradiation to determine the effects of microwave and He-Ne laser pretreatment on enzyme activities, and biophoton emission of cotyledon. Then: (i) changes in the activities of enzymes in I. indigotica cotyledon (such as amylase, transaminase, and proteinase) were measured to investigate the effects of He-Ne laser and microwave pretreatment; and (ii) biophoton emission was measured to determine the speed of cell division and metabolism. Results from these experiments indicated that: (i) the activities of amylase, transaminase, and proteinase of the cotyledon pretreated by HeNe laser and microwave were significantly increased; and (ii) the intensity of biophoton emission was enhanced significantly by He-Ne laser and microwave irradiation. These changes suggest that He-Ne laser and microwave pretreatment can improve the inner energy of seeds, lead to an enhancement of cotyledon enzymes, and speed up the metabolism of the cell, resulting in significantly increased biophoton emission.Moreover, the mechanism of action of the effects of laser and microwave radiation on the microcalorimetric parameters, enzyme activities, and biophoton emission of seeds is discussed on the basis of the results obtained.     

Microwave facilitation of methylatropine antagonism of central cholinomimetic drug effects

Pilocarpine-induced hypothermia and oxotremorine-induced tremors in mice are central cholinomimetic drug effects that are readily blocked by the muscarinic antagonist atropine. However, the quaternary ammonium derivative of atropine, methylatropine, is unable to block these cholinomimetic drug effects by virture of its inability to penetrate the blood-brain barrier (BBB) and blood-cerebral spinal fluid barrier (B-CSFB). Dose-response curves for pilocarpine and oxotremorine effects are not appreciably affected either by pretreatment with methylatropine (1.0 mg/kg) or by exposure to moderate-level microwave irradiation (2.45 GHz, 23.7 W/kg, CW, 10-min exposure). However, in mice receiving both the methylatropine pretreatment and microwave irradiation, the dose-response curves for both pilocarpine and oxotremorine effects were significantly shifted to the right, signifying a central anticholinergic action by methylatropine. These data indicate that a single acute exposure to a thermogenic level of microwave irradiation facilitates methylatropine antagonism of centrally mediated cholinomimetic drug effects. One possible explanation for this observation is that microwave radiation may enhance passage of quaternary ammonium compounds like methylatropine across the BBB and B-CSFB.     

Effect of Microwave and He-Ne Laser on Enzyme Activity and Biophoton Emission of Isatis indigotica Fort

The seed embryos of lsatis indigotica Fort were exposed to He-Ne laser (5.23 mW/mm^2, radiated for 5 min) and microwave ( 1.26 mW/mm^2, radiated for 8 s) irradiation to determine the effects of microwave and He-Ne laser pretreatment on enzyme activities, and biophoton emission of cotyledon. Then: (i) changes in the activities of enzymes in L indigotica cotyledon (such as amylase, transaminase, and proteinase) were measured to investigate the effects of He-Ne laser and microwave pretreatment; and (ii) biophoton emission was measured to determine the speed of cell division and metabolism. Results from these experiments indicated that: (i) the activities of amylase, transaminase, and proteinase of the cotyledon pretreated by HeNe laser and microwave were significantly increased; and (ii) the intensity of biophoton emission was enhanced significantly by He-Ne laser and microwave irradiation. These changes suggest that He-Ne laser and microwave pretreatment can improve the inner energy of seeds, lead to an enhancement of cotyledon enzymes, and speed up the metabolism of the cell, resulting in significantly increased biophoton emission. Moreover, the mechanism of action of the effects of laser and microwave radiation on the microcalorimetric parameters, enzyme activities, and biophoton emission of seeds is discussed on the basis of the results obtained.     

Thermodynamics and kinetics of joint action of antiviral agent tilorone and DMSO on model lipid membranes

Individual and joint action of two water-soluble drugs, DMSO and tilorone, on model l-α-dipalmitoylphosphatidylcholine (DPPC) membranes were studied in equilibrium and kinetic regimes by differential scanning calorimetry (DSC). For equilibrium experiments, the drugs were introduced during preparation of the model membrane. In kinetic studies, one of the drugs was added to the DPPC membrane already containing the other drug, and the effects of drug-membrane interactions were monitored in real-time regime. It was found that tilorone and DMSO had opposite effects on the membrane melting temperature, which were non-additive under joint introduction of these drugs. Analysis of kinetics of DSC profiles under drugs introduction allowed us to discriminate two processes in drug-membrane interactions with different characteristic times, i.e., drug sorption onto the membrane (minutes) and drug diffusion through stacks of lipid bilayers (hours). It was established that 0.1?mol% DMSO effectively enhanced membrane penetration for tilorone with the rate of tilorone diffusion being dependent upon the scheme of drugs administration. A model was proposed describing how sorption of a dopant onto lipid membrane could affect the membrane permeability for other dopants. Conditions were determined for enhancement of membrane permeability, as it was observed for DPPC/DMSO/tilorone system.     

Combination of pharmacological analgesics and microwave irradiation of an acupuncture point for suppression of visceral pain in mice: Role of the opioid and serotonergic cerebral systems

We studied suppression of pain-related reactions induced in mice by i.p. injection of 0.08 ml of a 2% solution of acetic acid using pharmacological analgesics (analgin and tramadol) combined with low-intensity microwave irradiation of an acupuncture point (AP) E-36 (frequency 30 to 300 GHz and power rate density 3·10⁻⁹ W/cm²). The respective effects were also observed under conditions of suppression of the functions of opioid and serotonergic cerebral systems using injections of, respectively, naloxone and DL-p-chlorophenylalanine (p-CPA). We found that antinociceptive effects provided by analgesics used in a 50% mean single dose in the combination with microwave irradiation of the AP were significantly more intense than those induced by isolated injection of analgesics used in both 50% and full mean single doses and isolated microwave irradiation of the AP E-36. After injections of naloxone, analgesic effects caused by the combined action of analgin and microwave irradiation of the AP were considerably smaller. At the same time, after injection of p-CPA, analgesic effects, provided by the combination of injection of pharmacological agents and microwave irradiation of the AP, weakened in the case of use of both analgesics. This was manifested in a significant increase in the total duration of pain-related behavioral reactions. Therefore, the studied analgesic effects observed in the examined animal groups are realized due to the involvement of the opioid and serotonergic cerebral systems. Neirofiziologiya/Neurophysiology, Vol. 39, No. 6, pp. 468–477, November–December, 2007.     

Microwave influence on the isolated heart function: II. Combined effect of radiation and some drugs

The combined effects of microwave radiation and some drugs were studied in an isolated frog auricle preparation. The experiments established that exposure to pulse-modulated 915 MHz microwaves for up to 40 min had no effect on either the rate or the amplitude of spontaneous auricle twitches, unless the average absorbed power was high enough to produce preparation heating. Treatment of the preparation with saline containing (0.6–3.0) 10^?5 M of propranolol or (0.5–1.5) 10^?7 M of atropine altered neither its pacemaker nor its contractile functions; these drugs also had no effect when they were combined with nonthermal microwave irradiation. Caffeine (1 mM) strongly increased the average heart power, which was calculated as the product of twitch rate and amplitude. The caffeine effect appeared to be significantly augmented (by about 15%, P<0.02) under exposure to burst-type pulsed microwaves (pulse width, 1.5 msec; pause, 2.5 msec; 8 pulses/burst, 16 bursts/s; average SAR, 8–10 W/kg). By itself, this modulation was not effective; the heating of the preparation and saline during exposure was approximately 0.1°C, which could not account for the detected changes. The experimental results demonstrate that caffeine treatment increases the microwave sensitivity of the frog auricle preparation and reveals primarily subthreshold, nonthermal microwave effect. © 1995 Wiley-Liss, Inc.     

The effect of 2450-MHz microwave radiation during microtubular polymerization in vitro

Exposure to 2450-MHz (cw) microwave radiation causes inhibition of cell division in intact cells and varied in vivo biological effects in both avian and mammalian species. Because these reported effects may result from alterations in the dynamics of microtubule formation, we studied the effects of simultaneous microwave exposure (2450 MHz, cw) during each of the three critical stages of the intracellar polymerization cycle. In addition, using circular dichroism spectroscopy, we studied the effect of microwave irradiation on the secondary structure of purified tubulin polypeptides. These studies were accomplished using specially constructed exposure systems that permit the continuous recording of turbidometric or circular dichroism measurements during simultaneous exposure to microwaves. The baseline turbidity of microtubular protein did not change under the influence of microwave radiation (20 or 200 mW/g SAR) and irradiation had no effect on the light-scattering properties of the depolymerized protein. EGTA-induced polymerization and cold-induced depolymerization patterns were also similar for both control and microwave-irradiated samples. The circular dichroism spectrum of purified tubulin also did not appear to be influenced by microwave irradiation, indicating a lack of effect on the protein secondary structure. The data suggest that the cellular effects of microwaves are not due to changes in microtubular proteins or their rate of polymerization.     

Estrogen potentiates the stereotypy induced by dopamine agonists in the rat

Repeated weekly treatment with 100 μg/kg of estradiol benzoate (EB) to ovariectomized female rats intensified the stereotypy induced by the dopamine agonists amphetamine and apomorphine. A similar effect on amphetamine-stereotypy was produced 48 hours after a single injection of 10 μg/kg of EB. The fact that EB failed to increase blood or brain levels of either ³H-amphetamine or ³H-apomorphine suggests that these behavioral effects were not due to altered peripheral drug metabolism or uptake into the brain. The enhancement of stereotypy produced by EB is viewed as one manifestation of a more complex modulatory influence of estrogen on DA function.     

Inactivation of Lactobacillus Bacteriophage PL-1 by Microwave Irradiation

The effect of microwave irradiation on the survival of bacteriophage PL-1, which is specific for Lactobacillus casei, was studied using a commercial 2,450 MHz microwave oven. The phages were inactivated by microwave irradiation according to almost first-order reaction kinetics. The rate of phage inactivation was not affected by the difference in the continuous or intermittent irradiation, nor by the concentrations of phages used, but was affected by the volume of phage suspensions, which prevented the loss of generated heat. Microwave irradiation of phage suspensions produced a number of ghost phages with empty heads, but fragmentation of the tail was hardly noticed. The breakage of phage genome DNA was primarily caused by the heat generated by microwave irradiation, whereas the phage DNA was not affected by the same temperature achieved by heat from outside. Thus we concluded that the phage-inactivating effect of microwave irradiation was mainly attributed to a thermal microwave effect, which was much stronger than a simple thermal exposure.     

The effect of microwave thermal denaturation on release properties of bovine serum albumin and gluten matrices

The purpose of this study was to compare the effects of denaturation by microwave irradiation on release properties of 2 physically different proteins. Matrices were prepared from water-soluble bovine serum albumin loaded with metoclopramide and sorbed with adequate amount of moisture were thermally denatured in a microwave oven. The release profile of the rather insoluble denatured albumin matrices followed the classical Fickian diffusion profile. The release rate was dependent on the degree of denaturation, which was highly dependent on the level of moisture originally absorbed by the albuminoidal matrices and the period of exposure to microwave energy. Consersely, attempts to reduce the rate of drug release through microwave irradiation of metoclopramide-loaded matrices prepared from water-insoluble gluten were futile. The denaturation process was shown to be limited to the relatively water-soluble protein core fraction, while aggregation between neighboring gluten proteins in the matrix was not achieved even in the presence of considerable amounts of sorbed water. Published: February 10, 2006     

Does microwave irradiation have other than thermal effects on histological staining of the mammalian CNS? A light microscopical study of microwave stimulated staining under isothermal conditions in man and rat.

The question whether or not microwave irradiation exerts other than thermal effects on histological staining is still a matter of controversy. The present study was undertaken to reveal or reject such a so far hypothetical non-thermal irradiation effect. A device was developed, which enables exposure of histological sections or tissue pieces to microwave irradiation under isothermal conditions, i.e. with synchronous removal of the internal heat produced. Three classical neuroanatomical staining methods were tested on human and rat CNS. As control, identical procedures were performed without simultaneous microwave irradiation. The experiments were performed at three different temperature levels ranging from 5 to 50 degrees C. In none of the cases studied was a light microscopically appreciable difference observed between the microwave and non-microwave versions of a stain at the same temperature. The hypothesis of a separate non-thermal effect of microwave irradiation on histological staining is therefore rejected.     

Alterations in amphetamine-induced locomotor activity and stereotypy after electrical stimulation of the nucleus accumbens and neostriatum

The exacerbation of the locomotor and stereotypic effects of amphetamine after repeated drug administration is well documented. To elaborate on the involvement of the nigrostriatal and mesolimbic dopamine (DA) systems in modulating behavioral sensitization, locomotor activity and the time spent engaged in repetitive stereotyped behaviors following systemic amphetamine injection were assessed after electrical stimulation of the nucleus accumbens and neostriatum. It was found that exposure to repeated sessions of high frequency, low current stimulation of the anteromedial neostriatum and nucleus accumbens significantly enhanced the locomotor excitation induced by administration of 3.0 mg/kg of amphetamine. Stereotypic behaviors were also modified as a function of electrical stimulation of these brain regions, with the development of a significant decrease in the duration of focused head and body movements corresponding to the facilitated locomotor effects of the drug. Taken together, these data provide additional evidence demonstrating the interdependent relationship between amphetamine-elicited locomotor activity and stereotypy, and were discussed in terms of a functional interaction between mesolimbic and nigrostriatal systems in determining the behavioral profile of amphetamine administration.     

Optimizing collagen antigen unmasking in paraffin-embedded tissues

In order to optimize collagen antigen unmasking in paraffin-embedded tissue sections, the effects of various fixatives and duration of fixation in relation to enzyme pretreatment and microwave irradiation for collagen antigen unmasking were studied. A streptavidin--biotin-- peroxidase complex method was used for the immunolocalization of type III and IV collagen antigens. Fixatives and fixation time had significant adverse effects on the immunoreactivity of the antigens. Enzyme pretreatment was found to be superior to microwave irradiation for collagen antigen unmasking. Fixation with paraformaldehyde required shorter enzyme pretreatment and yielded a more enhanced reaction than treatment with formalin and Bouin's fluid. The optimum conditions for type III and IV collagen unmasking were found to be fixation with 4% paraformaldehyde in 0.01 m phosphate-buffered saline, pH 7.4, for up to 3 weeks followed by enzyme pretreatment with 1 mg ml−1 pepsin in 0.01 n hydrochloric acid, pH 2.0, for 30 min (human tissues) or 60 min (rat tissues) at 37°C. It is concluded that collagen antigen unmasking by enzyme pretreatment in tissue sections fixed for a long period of time can be successful if appropriate enzyme(s) and incubation time(s) are employed with regard to the antigen under study and fixative and fixation time used for tissue preparation     

Optimizing collagen antigen unmasking in paraffin-embedded tissues

In order to optimize collagen antigen unmasking in paraffin-embedded tissue sections, the effects of various fixatives and duration of fixation in relation to enzyme pretreatment and microwave irradiation for collagen antigen unmasking were studied. A streptavidin--biotin-- peroxidase complex method was used for the immunolocalization of type III and IV collagen antigens. Fixatives and fixation time had significant adverse effects on the immunoreactivity of the antigens. Enzyme pretreatment was found to be superior to microwave irradiation for collagen antigen unmasking. Fixation with paraformaldehyde required shorter enzyme pretreatment and yielded a more enhanced reaction than treatment with formalin and Bouin's fluid. The optimum conditions for type III and IV collagen unmasking were found to be fixation with 4% paraformaldehyde in 0.01 m phosphate-buffered saline, pH 7.4, for up to 3 weeks followed by enzyme pretreatment with 1 mg ml−1 pepsin in 0.01 n hydrochloric acid, pH 2.0, for 30 min (human tissues) or 60 min (rat tissues) at 37°C. It is concluded that collagen antigen unmasking by enzyme pretreatment in tissue sections fixed for a long period of time can be successful if appropriate enzyme(s) and incubation time(s) are employed with regard to the antigen under study and fixative and fixation time used for tissue preparation     

微波辐射对肾小管上皮细胞的影响以及金雀异黄素对其保护作用

目的：观察微波辐照对人近端肾小管上皮细胞（HKC）的影响及金雀异黄素对其的保护作用。方法：HKC分为对照组、微波辐照组、金雀异黄素组（n=6）。金雀异黄素组在辐照前2 h用含30μmol/L金雀异黄素的DMEM培养基进行预培养。辐照后24 h留取上清进行乳酸脱氢酶（LDH）、β-N-乙酰氨基葡萄糖苷酶（NAG）活性及丙二醛（MDA）、超氧化物歧化酶（SOD）活性检测。Hoechst 33258染色观察细胞凋亡情况。结果：与对照组比较,微波辐照组上清NAG、LDH活性明显增加（P < 0.01）,金雀异黄素预处理组则较微波辐照组明显下降（P < 0.01）;微波辐照组上清活性也较对照组明显增加（P < 0.01）。Hoechst 33258染色显示,微波辐照可导致较多量的细胞凋亡,而应用金雀异黄素预处理后细胞凋亡的比例均大大减少。微波辐照可大大提高HKC细胞中的MDA含量,SOD活性降低（P< 0.01）,应用金雀异黄素预处理后MDA的含量无明显降低,SOD的活性明显增大（P < 0.01）。结论：微波辐照可导致人近端肾小管上皮细胞出现功能损伤,金雀异黄素对其具有一定的保护作用,可能与其减轻氧化应激损伤、减少细胞凋亡有关。     

Suppression of synthesis of serotonin: Effect on antinociception induced by microwave irradiation and analgesic remedies

We studied isolated and combined antinociceptive effects of analgesics (Analginum and Tramadol), as well as of microwave irradiation of an acupuncture point, in mice under conditions of suppression of synthesis of serotonin by an inhibitor of this synthesis, DL-p-chlorophenylalanine (p-CPA). Preliminary introduction of p-CPA weakened the antinociceptive effect of microwave irradiation and also moderated the combined effects of analgesics and microwave irradiation. Neirofiziologiya/Neurophysiology, Vol. 38, Nos. 5/6, pp. 498–499, September–December, 2006.     

The Raf/MEK/ERK pathway can govern drug resistance,apoptosis and sensitivity to targeted therapy

The effects of the Raf/MEK/ERK and PI3K/PTEN/Akt/mTOR signaling pathways on proliferation, drug resistance, prevention of apoptosis and sensitivity to signal transduction inhibitors were examined in FL/ΔAkt-1:ER*(Myr⁺) + ΔRaf-1:AR cells which are conditionally-transformed to grow in response to Raf and Akt activation. Drug resistant cells were isolated from FL/ΔAkt-1:ER*(Myr⁺) + ΔRaf-1:AR cells in the presence of doxorubicin. Activation of Raf-1, in the drug resistant FL/ΔAkt-1:ER*(Myr⁺) + ΔRaf-1:AR cells, increased the IC₅₀ for doxorubicin 80-fold, whereas activation of Akt-1, by itself, had no effect on the doxorubicin IC₅₀. However, Akt-1 activation enhanced cell proliferation and clonogenicity in the presence of chemotherapeutic drugs. Thus the Raf/MEK/ERK pathway had profound effects on the sensitivity to chemotherapeutic drugs, and Akt-1 activation was required for the long-term growth of these cells as well as resistance to chemotherapeutic drugs. The effects of doxorubicin on the induction of apoptosis in the drug resistant cells were enhanced by addition of either mTOR and MEK inhibitors. These results indicate that targeting the Raf/MEK/ERK and PI3K/Akt/mTOR pathways may be an effective approach for therapeutic intervention in drug resistant cancers that have mutations activating these cascades.