Fixed Versus Removable Microdialysis Probes for In Vivo Neurochemical Analysis: Implications for Behavioral Studies

Abstract: The levels of several neurochemicals, i.e., uric acid (UA), dopamine (DA), dihydroxyphenylacetic acid, and 5-hydroxyindoleacetic acid, collected daily from the rat striatum with either fixed or removable microdialysis probes for 7 days after surgery were compared. The implantation of the fixed cannula was followed by a 10-fold increase in the UA content in the dialysates collected from the first day after surgery onward and by a steady decrease in dihydroxyphenylacetic acid levels, whereas those of DA remained fairly stable. With the removable cannula system, only a smaller, transient increase in UA during the first 3 days after surgery was observed, with no change in DA or monoamine metabolites. The glial reaction around the cannula tracks was assessed by both quantitative histological techniques and measuring the glutamine levels in the dialysates collected at the time of surgery and 7 days later. Both the glial cell number and nuclear size, as well as the glutamine outflow, were considerably larger in the animals implanted with the fixed probes. It is, therefore, likely that the UA levels in the dialysate reflect the glial reaction to the probe. The suitability of the removable probe system for behavioral experiments involving repeated microdialysis sampling was illustrated in an experiment showing that the DA release in the nucleus accumbens of male rats assessed daily at postsurgery days 5–10 was virtually identical in three alternating sessions of sexual behavior as was the smaller release of this neurotransmitter detected during intervening nonsexual social interactions.     

Headache Treatment with Pulsing Electromagnetic Fields: A Literature Review

Pulsing electromagnetic field (PEMF) therapy may be a viable form of complementary and alternative medicine. Clinical applications include the treatment of fractures, wounds, and heart disease. More recent applications involve treatment of recurrent headache disorders. This paper reviews available studies investigating PEMF for headache management. Possible mechanisms for effects (neurochemical, electrophysical, and cardiovascular) are discussed. The available data suggest that PEMF treatment for headache merits further study. Suggestions for future research are provided.     

Perception of Risks from Electromagnetic Fields: Lessons for the Future

Technologies based on extremely highfrequency electromagnetic fields, inparticular in the terahertz region, arequite recent and new to the public. While anumber of advantages have been shown,especially in the biomedical area,biological effects and possible healthimplications have not been fullyinvestigated. The experience gained withelectromagnetic fields of lowerfrequencies, from ELF to microwaves,suggests that innovating technologies maycreate concern, or even fear, among thepublic for hypothetical health risks.Social research has shown that worries arerelated to the perception of risks by thepublic more than to their actual existence.Risk perception depends on several factors,many of which are relevant forelectromagnetic fields. They include lackof familiarity with the agent, difficultyin understanding interaction mechanisms,and uncertainty in scientific knowledge.Lessons learnt from the past lead torecommend that specific research onbiological effects of terahertz radiationbe started from the very beginning of thedevelopment of technological applicationsand that a continuous and effectivedialogue be established between thescientific community and the public.     

Interaction of Psychoactive Drugs with Exposure to Electromagnetic Fields

Evidence has previously been presented to show that the dopamine (DA) and opiate systems of the brain, in part, mediate the effects of exposure to electromagnetic (EM) fields. The hypothesis was broadened to predict that low-intensity EM energy exposure would interact with naloxone, chlor-diazepoxide and haloperidol treatment in tests using analgesia and stereotypic behavior. In the set of experiments reported here, the data obtained indicated there were such interactions.

One implication of the dopamine-opiate hypothesis is that EM energy exposure would likely affect the hypothalamic set-point for body temperature regulation. Substantial evidence for this view now exits. This theory and data indicate that body temperature rise with exposure is likely to be a consequence of the energy affecting the dopamine-opiate systems of the brain.     

Rapid Stereoselective Hydrolysis of (+)-Cocaine in Baboon Plasma Prevents Its Uptake in the Brain: Implications for Behavioral Studies

The naturally occurring enantiomer of cocaine, (–)-cocaine, has been previously labeled with ¹¹C on the N-methyl group and used in conjunction with positron emission tomography to show that cocaine is rapidly taken up in the striata of human and baboon brain. In the present study, the behaviorally inactive (+)-cocaine was similarly labeled, with a view to its use for measuring the nonspecific binding of cocaine. No brain uptake was seen, although transport of cocaine into the brain is not expected to be stereoselective. The explanation for the lack of uptake was determined to be very rapid metabolism of (+)-cocaine in the blood. By 30s after administration of labeled (+)-cocaine, it was undetectable in plasma. In vitro studies demonstrated that (+)-cocaine is 50% debenzoylated to (+)-ecgonine methyl ester within 5 s of exposure to baboon plasma but not to washed erythrocytes. The hydrolysis of (–)-cocaine is at least 1,000 times slower. Serum butyrylcholinesterase (EC 3.1.1.8) appears to be responsible for this hydrolysis, as evidenced by its inhibition by physostigmine and catalysis by commercially available pseudocholinesterase from horse and human blood.     

Pulsed Electromagnetic Fields and Normal Chick Development

The effects of two clinical-type signals of pulsed electromagnetic fields (PEW) were tested on a large sample of developing chick embryos by two independent laboratories. The embryos were examined after seven days of incubation to avoid the early stages when most spontaneous abnormalities occur. The data were pooled and analyzed statistically. Neither signal, when administered continuously for the first seven days of development, or for the first 24 hours, produced a significant increase in the incidence of malformations. These results are discussed in relation to those of other investigators claiming PEW-related effects on normal development.     

Pseudoarthrosis Treatment with Cancellous Bone Grafts and Low-Frequency Pulsing Electromagnetic Fields

We report our experience in the use of cancellous bone grafts and stimulation with pulsing electromagnetic fields (PEMFs) in the treatment of acquired pseudarthrosis in 28 patients, including 17 patients with active infections. Stimulation was begun immediately after the grafting, and all patients healed in an average time of 4 months.     

Reinterpreting Behavioral Receptive Fields: Lightness Induction Alters Visually Completed Shape

Background

A classification image (CI) technique has shown that static luminance noise near visually completed contours affects the discrimination of fat and thin Kanizsa shapes. These influential noise regions were proposed to reveal “behavioral receptive fields” of completed contours–the same regions to which early cortical cells respond in neurophysiological studies of contour completion. Here, we hypothesized that 1) influential noise regions correspond to the surfaces that distinguish fat and thin shapes (hereafter, key regions); and 2) key region noise biases a “fat” response to the extent that its contrast polarity (lighter or darker than background) matches the shape''s filled-in surface color.

Results

To test our hypothesis, we had observers discriminate fat and thin noise-embedded rectangles that were defined by either illusory or luminance-defined contours (Experiment 1). Surrounding elements (“inducers”) caused the shapes to appear either lighter or darker than the background–a process sometimes referred to as lightness induction. For both illusory and luminance-defined rectangles, key region noise biased a fat response to the extent that its contrast polarity (light or dark) matched the induced surface color. When lightness induction was minimized, luminance noise had no consistent influence on shape discrimination. This pattern arose when pixels immediately adjacent to the discriminated boundaries were excluded from the analysis (Experiment 2) and also when the noise was restricted to the key regions so that the noise never overlapped with the physically visible edges (Experiment 3). The lightness effects did not occur in the absence of enclosing boundaries (Experiment 4).

Conclusions

Under noisy conditions, lightness induction alters visually completed shape. Moreover, behavioral receptive fields derived in CI studies do not correspond to contours per se but to filled-in surface regions contained by those contours. The relevance of lightness to two-dimensional shape completion supplies a new constraint for models of object perception.     

Reduced Dependence of Crested Ibis on Winter-Flooded Rice Fields: Implications for Their Conservation

The Crested Ibis Nipponia nippon was once thought to be extinct in the wild until seven birds were discovered in a remote mountain village in China in 1981. Studies suggested that winter-flooded rice fields play an essential role in nest site selection by the Crested Ibis and hence in their survival. Considerable efforts were therefore made to conserve the winter-flooded rice fields, but these have caused conflicts between the agricultural and conservation communities. The population and geographical range of the wild Crested Ibis has expanded greatly since 1981, but there is no spatial information on the winter-flooded rice fields, nor on the current association of nest sites and winter-flooded rice fields. We mapped winter-flooded rice fields across the entire current range of Crested Ibis using innovative remote sensing and geographical information systems (GIS) techniques. The spatial relationships between the nest site clusters and winter-flooded rice fields were quantified using Ward''s hierarchical clustering method and Ripley''s K-function. We show that both have significantly clumped distribution patterns and that they are positively associated. However, the dependence of Crested Ibis on the winter-flooded rice fields varied significantly among the nest site clusters and has decreased over the years, indicating the absence of winter-flooded rice fields is not constraining their recovery and population expansion. We therefore recommend that efforts should be made to protect the existing winter-flooded rice fields and to restore the functionality of natural and semi-natural wetlands, to encourage both in-situ conservation and the re-introduction of the Crested Ibis. In addition, we recommend that caution should be exercised when interpreting the habitat requirements of species with a narrow distribution, particularly when that interpretation is based only on their current habitat.     

Magnetic Fields in Earth-like Exoplanets and Implications for Habitability around M-dwarfs

We present estimations of dipolar magnetic moments for terrestrial exoplanets using the Olson & Christiansen (EPS Lett 250:561–571, 2006) scaling law and assuming their interior structure is similar to Earth. We find that the dipolar moment of fast rotating planets (where the Coriolis force dominates convection in the core), may amount up to ~80 times the magnetic moment of Earth, M _⊕ , for at least part of the planets’ lifetime. For slow rotating planets (where the force of inertia dominates), the dipolar magnetic moment only reaches up to ~1.5 M _⊕ . Applying our calculations to confirmed rocky exoplanets, we find that CoRoT-7b, Kepler-10b and 55 Cnc e can sustain dynamos up to ~18, 15 and 13 M _⊕ , respectively. Our results also indicate that the magnetic moment of rocky exoplanets not only depends on rotation rate, but also on their formation history, thermal state, age, composition, and the geometry of the field. These results apply to all rocky planets, but have important implications for the particular case of planets in the Habitable Zone of M-dwarfs.     

Treatment of Congenital Pseudarthrosis with Endomedullary Nail and Low Frequency Pulsing Electromagnetic Fields: A Controlled Study

We report our experience in the treatment of congenital pseudarthrosis with pulsing electromagnetic fields in a controlled study. Both the stimulated and control groups received the same surgical procedure: excision of the pseudarthrosis site, reduction and fixation by intramedullary nail. Stimulation with electromagnetic fields was begun within 3 days after surgery. The orthopaedic treatment was the same for all patients. The stimulation lasted up to 12 months. All patients were followed at least 24 months after the surgery. The data showed that the surgical approach in association with pulsing electromagnetic-field stimulation gave significantly better results than surgery alone.     

In Vitro Evaluation of the Effects of Electromagnetic Fields used for Bone Healing

The use of therapeutic electromagnetic fields (EMF) for bone healing has positive clinical effects but may have adverse biologic effects. For this reason, EMF exposure has been repeatedly investigated to exclude the possibility of genotoxic effects and tumour risk. This paper describes the effects of EMFs on cell cultures. We analyzed the effects of EMF (28 gauss, 75 Hz) on growth and metabolic activities in four different cell types: L929 fibro-blasts, osteoblast-like HOS/TE85 cells, human lymphocytes, and rabbit chondrocytes. We found no cytotoxic or mutagenic effects on cultures exposed to EMF compared with unexposed controls. Results of cell proliferation showed a statistically significant increase for all cultures exposed to EMF with respect to controls (L929 +45%, p = 0.002; HOS/TE85 +32%, p = 0.001; chondrocytes +40%, p = 0.0003; lymphocytes +39%, p = 0.0002). Biochemical and enzymatic tests gave different results, depending on cell types: all tested values were increased after EMF exposure, even if only some of them reached statistical significance (total proteins: HOS/TE85 p = 0.004, chondrocytes p = 0.003; alkaline phosphatase: L929p = 0.0003, HOS/RE85 p = 0.0001, chondrocytes p = 0.009, lymphocytes p = 0.006; lactate dehydrogenase: chondrocytes p = 0.0002, lymphocytes p = 0.0005). Biochemical and enzymatic tests and cell proliferation results suggest a more active metabolism in cartilage and bone cells after EMF exposure. These effects could be relevant for bone healing in clinical practice.     

The Effects of Electromagnetic Fields on Human Health:Recent Advances and Future

The potential of clectromagnetic ficlds(EMFs)for discase treatment and health enhancement has been actively pursued over the recent decades.This review first provides a general introduction about natural EMFs and related biological effcts.Then the recent progress on the EMF treatment of some common diseases(such as cancer,diabetes,wound healing and neurological diseases,etc.)has been carefully reviewed and summarized.Yet,the blindness on the selection of therapeutic EMF parameters still hinders the broad application ofEMF therapy.Moreover,the unclear mechanism of EMF function and poor reproducibility of experimental rcsults also remain big challenges in the field of bioclectromagnetics.Bionics is a useful methodology that gains inspiration from nature to serve human life and industry.We have discussed the feasibility of applying bionic approach on the selection of the rapeutic EMFs,which is based on thefindings of natural EMFs.Finally,we advocate that the detailed information of EMFs and biological samples should be thoroughly reorded in future research and reported in publications.In addition,the publication of studies with negative results should also be allowed.     

Histopathological and Ultrastructural Studies on the Effects of Electromagnetic Fields on the Liver of Preincubated White Leghorn Chicken Embryo

There are several reports indicating a linkage between exposures to 50–60 Hz electromagnetic fields and abnormalities in the early stages of embryonic development of chicken embryos. The present study was designed to demonstrate whether electromagnetic fields could be an environmental factor invoking histopathological and ultra-structural changes in livers of preincubated chicken embryos exposed to EMFs. Following other researchers and our previous results from different groups of Developmental Biology at the Animal Sciences, Faculty of Biological Sciences, Shahid-Beheshti University, effects of most effective intensities (1.33, 2.66, 5.52, and 7.32 mT) of electromagnetic fields (EMFs, 50 Hz ) on livers of pre-incubated white leghorn chicken embryos were investigated . 150 healthy, fresh, and fertilized eggs (55–65 gr) were divided into 6 groups of experimental(1–4, n = 30), control (n = 60), and sham (n = 50). Experimental eggs (inside coil) were exposed to 4 different intensities (1.33, 2.66, 5.52, and 7.32 mT). Sham groups were located inside same coil, with no exposure, for 24 h before incubation. Control, sham, and experimental groups (1–4) were then incubated in an incubator (38 ± 0.5°C, 60% humidity) for 17 days. At the end of this period, livers of experimental, sham, and control groups were processed for light and transmission electrom microscopes (TEM and SEM) studies. So, livers of 17-day old chicken embryos were removed by C-sections, fixed in formalin 10%, stained with H&E and reticulin, and studied under light microscope. Others were prepared for electron microscopes (TEM and SEM) investigations. Morphological observations indicated exencephalic embryos, embryos with asymmetrical faces, crossed beak, shorter upper beak, deformed hind limbs, gastroschesis, anophthalmia, and microphthalmia. H&E and reticulin stainings, TEMS, and SEMs studies indicated EMFs would create hepato-cytes with fibrotic bands, severe steatohepatitis, vacuolizations, swollen and extremely electron-dense mitochondria, reduced invisible cristae, crystalized mitochondria with degenerated cristae, myelin-like figures, macrophages engulfing adjacent cells, dentated nuclei, nuclei with irregular envelopes, degenerated hepatocytes, abnormal lipid accumulations, lipid droplets pushing hepatocytes' nuclei to the corner of the cells, abundant cellular infiltrations cellular infiltrations inside sinusoid and around central veins, disrupted reticulin plexus, and release of chromatin into cytosol,, with partially regular water layers. An elevated oxyradical generation and, subsequentely, cell membrane disruptions were the reasons for electromagnetic fields inducing cell damages.     

Ovarian Carcinoma Subtypes Are Different Diseases: Implications for Biomarker Studies

Background

Although it has long been appreciated that ovarian carcinoma subtypes (serous, clear cell, endometrioid, and mucinous) are associated with different natural histories, most ovarian carcinoma biomarker studies and current treatment protocols for women with this disease are not subtype specific. With the emergence of high-throughput molecular techniques, distinct pathogenetic pathways have been identified in these subtypes. We examined variation in biomarker expression rates between subtypes, and how this influences correlations between biomarker expression and stage at diagnosis or prognosis.

Methods and Findings

In this retrospective study we assessed the protein expression of 21 candidate tissue-based biomarkers (CA125, CRABP-II, EpCam, ER, F-Spondin, HE4, IGF2, K-Cadherin, Ki-67, KISS1, Matriptase, Mesothelin, MIF, MMP7, p21, p53, PAX8, PR, SLPI, TROP2, WT1) in a population-based cohort of 500 ovarian carcinomas that was collected over the period from 1984 to 2000. The expression of 20 of the 21 biomarkers differs significantly between subtypes, but does not vary across stage within each subtype. Survival analyses show that nine of the 21 biomarkers are prognostic indicators in the entire cohort but when analyzed by subtype only three remain prognostic indicators in the high-grade serous and none in the clear cell subtype. For example, tumor proliferation, as assessed by Ki-67 staining, varies markedly between different subtypes and is an unfavourable prognostic marker in the entire cohort (risk ratio [RR] 1.7, 95% confidence interval [CI] 1.2%–2.4%) but is not of prognostic significance within any subtype. Prognostic associations can even show an inverse correlation within the entire cohort, when compared to a specific subtype. For example, WT1 is more frequently expressed in high-grade serous carcinomas, an aggressive subtype, and is an unfavourable prognostic marker within the entire cohort of ovarian carcinomas (RR 1.7, 95% CI 1.2%–2.3%), but is a favourable prognostic marker within the high-grade serous subtype (RR 0.5, 95% CI 0.3%–0.8%).

Conclusions

The association of biomarker expression with survival varies substantially between subtypes, and can easily be overlooked in whole cohort analyses. To avoid this effect, each subtype within a cohort should be analyzed discretely. Ovarian carcinoma subtypes are different diseases, and these differences should be reflected in clinical research study design and ultimately in the management of ovarian carcinoma.     

Electromagnetic Fields Induced by Helmholtz Aiding Coils Inside Saline-Filled Boundaries

The use of Helmholtz aiding coils to induce small electromagnetic (EM) fields in living tissue for both medical and research purposes has become quite common. While much progress has been made in showing that these induced EM signals can cause a variety of effects in tissues and individual cells, a satisfactory explanation of how the effects occur or how the EM signals couple to the tissue has not yet emerged. To address the latter problem adequately, it becomes necessary to know the spatial distribution of the induced fields inside a given set of boundaries. This paper examines the situation used for much in vitro research where a cylindrical culture dish is filled with a conducting solution and placed between the Helmholtz coils. Two cases are considered. The first assumes that the coils are above and below the culture dish (the planes of the coils are parallel with the top and bottom of the dish); the second assumes that the planes of the coils are parallel with the sides of the dish. A closed form solution is obtained for both cases, and it is shown that the induced EM field distribution is markedly different for the two cases.     

Effect of Electromagnetic Fields on Two Calcium Dependent Biological Systems

Weak pulsing electromagnetic fields interfered with lymphocyte blastogenesis and thromboxane release by polymorphonuclear leukocytes, as measured with %-thymidine incorporation and radioimmunoassay, respectively. Both cellular systems are calcium-dependent. Electromagnetic exposure significantly Inhibits both DNA synthesis and uptake in transformed lymphocytes. The effect on DNA synthesis seems to be synergistic with that of verapamil, a calcium blocker.