Mesencephalic Electrical Stimulation Reduces Neuroinflammation after Photothrombotic Stroke in Rats by Targeting the Cholinergic Anti-Inflammatory Pathway.

Inflammation is crucial in the pathophysiology of stroke and thus a promising therapeutic target. High-frequency stimulation (HFS) of the mesencephalic locomotor region (MLR) reduces perilesional inflammation after photothrombotic stroke (PTS). However, the underlying mechanism is not completely understood. Since distinct neural and immune cells respond to electrical stimulation by releasing acetylcholine, we hypothesize that HFS might trigger the cholinergic anti-inflammatory pathway via activation of the α7 nicotinic acetylcholine receptor (α7nAchR). To test this hypothesis, rats underwent PTS and implantation of a microelectrode into the MLR. Three hours after intervention, either HFS or sham-stimulation of the MLR was applied for 24 h. IFN-γ, TNF-α, and IL-1α were quantified by cytometric bead array. Choline acetyltransferase (ChAT)+ CD4+-cells and α7nAchR+-cells were quantified visually using immunohistochemistry. Phosphorylation of NFĸB, ERK1/2, Akt, and Stat3 was determined by Western blot analyses. IFN-γ, TNF-α, and IL-1α were decreased in the perilesional area of stimulated rats compared to controls. The number of ChAT+ CD4+-cells increased after MLR-HFS, whereas the amount of α7nAchR+-cells was similar in both groups. Phospho-ERK1/2 was reduced significantly in stimulated rats. The present study suggests that MLR-HFS may trigger anti-inflammatory processes within the perilesional area by modulating the cholinergic system, probably via activation of the α7nAchR.

Near-infrared light is neuroprotective in a monkey model of Parkinson disease.

OBJECTIVE: To examine whether near-infrared light (NIr) treatment reduces clinical signs and/or offers neuroprotection in a subacute 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) monkey model of Parkinson disease. METHODS: We implanted an optical fiber device that delivered NIr (670 nm) to the midbrain of macaque monkeys, close to the substantia nigra of both sides. MPTP injections (1.5-2.1mg/kg) were made over a 5- to 7-day period, during which time the NIr device was turned on. This was then followed by a 3-week survival period. Monkeys were evaluated clinically (eg, posture, bradykinesia) and behaviorally (open field test), and their brains were processed for immunohistochemistry and stereology. RESULTS: All monkeys in the MPTP group developed severe clinical and behavioral impairment (mean clinical scores = 21-34; n = 11). By contrast, the MPTP-NIr group developed much less clinical and behavioral impairment (n = 9); some monkeys developed moderate clinical signs (mean scores = 11-15; n = 3), whereas the majority--quite remarkably--developed few clinical signs (mean scores = 1-6; n = 6). The monkeys that developed moderate clinical signs had hematic fluid in their optical fibers at postmortem, presumably limiting NIr exposure and overall clinical improvement. NIr was not toxic to brain tissue and offered neuroprotection to dopaminergic cells and their terminations against MPTP insult, particularly in animals that developed few clinical signs. INTERPRETATION: Our findings indicate NIr to be an effective therapeutic agent in a primate model of the disease and create the template for translation into clinical trials.

Chronic exposure to broadband noise at moderate sound pressure levels spatially shifts tone-evoked responses in the rat auditory midbrain.

Noise-induced hearing disorders are a significant public health concern. One cause of such disorders is exposure to high sound pressure levels (SPLs) above 85 dBA for eight hours/day. High SPL exposures occur in occupational and recreational settings and affect a substantial proportion of the population. However, an even larger proportion is exposed to more moderate SPLs for longer durations. Therefore, there is significant need to better understand the impact of chronic, moderate SPL exposures on auditory processing, especially in the absence of hearing loss. In this study, we applied functional magnetic resonance imaging (fMRI) with tonal acoustic stimulation on an established broadband rat exposure model (65 dB SPL, 30 kHz low-pass, 60 days). The auditory midbrain response of exposed subjects to 7 kHz stimulation (within exposure bandwidth) shifts dorsolaterally to regions that typically respond to lower stimulation frequencies. This shift is quantified by a region of interest analysis that shows that fMRI signals are higher in the dorsolateral midbrain of exposed subjects and in the ventromedial midbrain of control subjects (p<0.05). Also, the center of the responsive region in exposed subjects shifts dorsally relative to that of controls (p<0.05). A similar statistically significant shift (p<0.01) is observed using 40 kHz stimulation (above exposure bandwidth). The results suggest that high frequency midbrain regions above the exposure bandwidth spatially expand due to exposure. This expansion shifts lower frequency regions dorsolaterally. Similar observations have previously been made in the rat auditory cortex. Therefore, moderate SPL exposures affect auditory processing at multiple levels, from the auditory cortex to the midbrain.

Visualization and quantification of cerebral metabolic fluxes of glucose in awake mice.

Biotransformation of glucose in organs includes multiple pathways, while quantitative evaluation of percentages of its utilization for individual pathways and their spatial heterogeneity in vivo remain unknown. Imaging MS (IMS) and metabolomics combined with a focused microwave irradiation for rapidly fixing tissue metabolism allowed us to quantify and visualize metabolic fluxes of glucose-derived metabolites in the mouse brain in vivo. At 15 min after the intraperitoneal injection of (13) C6 -labeled glucose, the mouse brain was exposed to focused microwave irradiation, which can stop brain metabolism within 1 s. Quantification of metabolic intermediates containing (13) C atoms revealed that a majority of the (13) C6 -glucose was diverted into syntheses of glutamate, lactate, and uridine diphosphate (UDP)-glucose. IMS showed that regions rich in glutaminergic neurons exhibited a large signal of (13) C2 -labeled glutamate. On the other hand, the midbrain region was enriched with an intensive (13) C6 -labeled UDP-glucose signal, suggesting an active glycogen synthesis. Collectively, application of the current method makes it possible to examine the fluxes of glucose metabolism in a region-specific manner.

Variations in amino acid neurotransmitters in some brain areas of adult and young male albino rats due to exposure to mobile phone radiation.

BACKGROUND AND OBJECTIVES: Mobile phone radiation and health concerns have been raised, especially following the enormous increase in the use of wireless mobile telephony throughout the world. The present study aims to investigate the effect of one hour daily exposure to electromagnetic radiation (EMR) with frequency of 900 Mz (SAR 1.165 w/kg, power density 0.02 mW/cm2) on the levels of amino acid neurotransmitters in the midbrain, cerebellum and medulla of adult and young male albino rats. MATERIALS AND METHODS: Adult and young rats were divided into two main groups (treated and control). The treated group of both adult and young rats was exposed to EMR for 1 hour daily. The other group of both adult and young animals was served as control. The determination of amino acid levels was carried out after 1 hour, 1 month, 2 months and 4 months of EMR exposure as well as after stopping radiation. RESULTS: Data of the present study showed a significant increase in both excitatory and inhibitory amino acids in the cerebellum of adult and young rats and midbrain of adult animals after 1 hour of EMR exposure. In the midbrain of adult animals, there was a significant increase in glycine level after 1 month followed by significant increase in GABA after 4 months. Young rats showed significant decreases in the midbrain excitatory amino acids. In the medulla, the equilibrium ratio percent (ER%) calculations showed a state of neurochemical inhibition after 4 months in case of adult animals, whereas in young animals, the neurochemical inhibitory state was observed after 1 month of exposure due to significant decrease in glutamate and aspartate levels. This state was converted to excitation after 4 months due to the increase in glutamate level. CONCLUSION: The present changes in amino acid concentrations may underlie the reported adverse effects of using mobile phones.

Differential numbers of foci of lymphocytes within the brains of Lewis rats exposed to weak complex nocturnal magnetic fields during development of experimental allergic encephalomyelitis.

To discern if specific structures of the rat brain contained more foci of lymphocytes following induction of experimental allergic encephalomyelitis and exposures to weak, amplitude-modulated magnetic fields for 6 min once per hour during the scotophase, the residuals between the observed and predicted values for the numbers of foci for 320 structures were obtained. Compared to the brains of sham-field exposed rats, the brains of rats exposed to 7-Hz 50 nT (0.5 mG) amplitude-modulated fields showed more foci within hippocampal structures and the dorsal central grey of the midbrain while those exposed to 7-Hz 500 nT (5 mG) fields showed greater densities within the hypothalamus and optic chiasm. The brains of rats exposed to either the 50 nT or 500 nT amplitude-modulated 40-Hz fields displayed greater densities of foci within the midbrain structures related to rapid eye movement. Most of the enhancements of infiltrations within the magnetic field-exposed rats occurred in structures within periventricular or periaqueductal regions and were both frequency- and intensity-dependent. The specificity and complexity of the configurations of the residuals of the numbers of infiltrated foci following exposures to the different fields suggest that the brain itself may be a "sensory organ" for the detection of these stimuli.

Intracranial application of near-infrared light in a hemi-parkinsonian rat model: the impact on behavior and cell survival.

OBJECT The authors of this study used a newly developed intracranial optical fiber device to deliver near-infrared light (NIr) to the midbrain of 6-hydroxydopamine (6-OHDA)-lesioned rats, a model of Parkinson's disease. The authors explored whether NIr had any impact on apomorphine-induced turning behavior and whether it was neuroprotective. METHODS Two NIr powers (333 nW and 0.16 mW), modes of delivery (pulse and continuous), and total doses (634 mJ and 304 J) were tested, together with the feasibility of a midbrain implant site, one considered for later use in primates. Following a striatal 6-OHDA injection, the NIr optical fiber device was implanted surgically into the midline midbrain area of Wistar rats. Animals were tested for apomorphine-induced rotations, and then, 23 days later, their brains were aldehyde fixed for routine immunohistochemical analysis. RESULTS The results showed that there was no evidence of tissue toxicity by NIr in the midbrain. After 6-OHDA lesion, regardless of mode of delivery or total dose, NIr reduced apomorphine-induced rotations at the stronger, but not at the weaker, power. The authors found that neuroprotection, as assessed by tyrosine hydroxylase expression in midbrain dopaminergic cells, could account for some, but not all, of the observed behavioral improvements; the groups that were associated with fewer rotations did not all necessarily have a greater number of surviving cells. There may have been other "symptomatic" elements contributing to behavioral improvements in these rats. CONCLUSIONS In summary, when delivered at the appropriate power, delivery mode, and dosage, NIr treatment provided both improved behavior and neuroprotection in 6-OHDA-lesioned rats.

Sympathetically induced renal vasoconstriction during stimulation of mesencephalic locomotor region in rats.

Central command, which is a neural drive originating in the brain during exercise, regulates the sympathetic nervous system and evokes cardiovascular responses to exercise. To examine the role of the central command on sympathetic regulation of renal circulation, we compared responses in renal cortical blood flow and vascular conductance to electrical stimulation of mesencephalic locomotor region (MLR) for 30 s in decerebrate and paralyzed rats between renal nerves that were intact (n=8) and denervated (n=8). In rats with renal nerves intact, stimulation of the MLR at 40 microA current intensity significantly (p<0.05) decreased renal cortical blood flow (-17+/-5%, means+/-S.E.M.) and vascular conductance (-43+/-4%) and the decrease in renal vascular conductance was dependent on current intensity (between 20 and 60 microA). In renal denervated rats, in contrast, there were no significant changes in either renal cortical blood flow or vascular conductance during stimulation at all current intensities. In a subset of rats (n=8), the response in renal sympathetic nerve activity to 30 s stimulation of the MLR was investigated. Stimulation of the MLR significantly increased renal sympathetic nerve activity (+57+/-14% at 40 microA) and the response was dependent on current intensity (between 20 and 60 microA). These data provide evidence that central command induces renal vasoconstriction by increasing sympathetic activity, depending on central command intensity.

[Fulminating midbrain irradiation injury of pediatric brain tumor].

We report two children with post radiation midbrain damage causing severe neurological symptoms. A twelve-year-old boy with a four year history of hydrocephalus was diagnosed with tectal glioma, which endoscopic biopsy revealed to be low grade. He underwent gamma knife radiation surgery (central 24 Gy/peripheral 12 Gy). Two months later bilateral ptosis followed by total oculomotor palsy and drowsiness developed. Despite pulsed-steroid therapy the tumor size increased up to 4.6 times in volume. The tumor was totally removed and was diagnosed as an early delayed radiation reaction pathologically. His symptoms disappeared except for a slight upper gaze palsy. The second patient was a six-year-old girl with a medulloblastoma. Following total resection and a VP shunt she received conventional radiation therapy along with chemotherapy. After the final irradiation she became comatose (JCS II-2) and MRI revealed diffuse midbrain damage with acute aqueduct obstruction, which recovered in two weeks. Reports of irradiation injuries of the midbrain in childhood are rare but it should be considered as a possible cause of fulminant symptoms requiring emergency treatment. Because of midbrain anatomical complexity, midbrain radiation therapy requires great care, especially in children.

Late ultrastructural changes in neuronal mitochondria after lonizing radiation of the brain.

Brain tissue was examined for morphological changes at 12 and 16 months after focal irradiation of the brain in female mice. Irradiation was performed with deuteron beams derived from the 60-inch Brookhaven cyclotron at a rate of 1925 rad second -minus 1 though an anti-Bragg device which produced uniform exposure. Experimental animals received a dose of 10,000 rad, covering a 9 by 5 mm area over the skull and extending into the brain from a depth of about 2 mm. An additional group of animals served as sham-irradiated controls. Histological studies revealed extensive cell loss, vacuolation, and prominent vascular changes in irradiated regions of the brain at 16 months post-irradiation. Ultrastructural examination of brain tissue at 12 and 16 months after irradiation revealed the presence of unusual elongated mitochondria with parallel arrays of cristae. Altered mitochondria were more prevalent at the longer post-irradiation interval.

Radiotherapy of midbrain and brainstem tumours in children.

From 1984 to 1987 in the First Radiotherapy Department in the Centre of Oncology in Warsaw, 11 children with midbrain (group I) and 14 with brainstem (group II) tumours were treated. In 4 cases diagnostic biopsy was performed and in 21 diagnosis was established by CT scan. All children were treated with megavoltage radiotherapy with a Co-60 unit. The initial radiotherapy treatment volume was determined from CT scan and was subsequently adapted to include whole brain or whole cranio-spinal axis, depending on the response to treatment. Improvement or stabilization of disease in 23/25 (92%) of cases was observed. Total survival, longer than 3 years in 14/25 (56%) was observed, while 9/11 (82%) survived greater than 3 years (NED) in group I, and in 4/14 (28%) in group II. Ninety two percent of living children have normal school life, with minimal or no neurological defects.

Induction of apoptosis in cultured midbrain cells from embryonic mice.

Midbrain cells from embryonic mice were cultured at a high cell density for 4 days to observe proliferation and differentiation. Results indicate that UVC radiation caused dose-dependent inhibition of their proliferation and differentiation. Inhibitory effects of UVC radiation (per unit dose) were greater on cell differentiation than on proliferation. The percentage of apoptotic cells in the groups irradiated with 15 J/m2 became significantly higher than the control level 2 h after irradiation. The percentage of apoptotic cells increased with time in a dose-associated fashion and reached a maximum at about 7.5 h after irradiation. Then it decreased with time and returned to the control level at about 30 h after irradiation. Metabolic inhibitors could postpone occurrence of intermediate apoptosis (>2 h, but <20 h) if they were added at suitable times after irradiation but failed to prevent a delayed apoptosis (>24 h). Results showed that UVC radiation induced intermediate apoptosis which could be postponed by metabolic inhibitors, and that UVC could also induce delayed apoptosis with the addition of metabolic inhibitors. The result that UVC radiation could induce intermediate apoptosis in the midbrain cells was different from previous data obtained using cells of an established cell line. The implications of the results for the understanding of the mechanism involved are discussed.

Stimulation in the rat of a nerve fiber bundle by a short UV pulse from an excimer laser.

A bundle of central nervous fibers was excited in the rat with a short pulse (40 ns) of UV light produced by an excimer laser. Evoked responses were recorded in the thalamic ventralis posterior nucleus after stimulation of the medial lemniscus or the cuneate bundle in the spinal cord. The effects of electrical and optical fiber applied UV stimulation were compared in both cases. At threshold, the latency for the UV light stimulation was slightly longer than for electrical stimulation. The excitation threshold was 0.9 J/cm2, very close to the UV photoablation threshold (order of 1 J/cm2). The intermediary events mediating the light excitation are discussed.

Histology of the irradiated rat brain in the first post-irradiation year.

The histology of the brains of rats killed were studied, following various doses of 250 kV X rays. The majority of rats which were killed before 200 days showed no damage to the mid-brain and cerebral hemispheres. However, the trigeminal and facial nerves exhibited necrosis, the origin of which was probably in the central nervous tissue of the transitional region between central and peripheral nerve systems. The result of these lesions was atrophy of the masticatory muscles and a consequent difficulty in feeding. Rats surviving beyond 200 days post-irradiation exhibited focal necrosis within central white matter regions which resembled, both in form and location, those described in previous reports for rats.

[Effect of static magnetic field on development toxicity of rat embryonic midbrain neurons cells].

OBJECTIVE: To explore the effect of static magnetic field (SMF) on the differentiation and proliferation of rat embryonic midbrain neurons cells. METHODS: The micromass culture of rat embryonic midbrain neurons cell was applied to study the effect of varieties of SMF (1.0, 10.0, 50.0, 100.0, 200.0 mT) and FACS. RESULTS: SMF inhibited the differentiation of the cell without affecting cell proliferation. The concentration of 50% inhibition of cell differentiation (ICD(50)) was 25 mT. The concentration of 50% inhibition of cell differentiation (IVD(50)) was 45 mT. CONCLUSIONS: The inhibition of SMF on embryonic midbrain neurons cells may be associated with impact of protein syntheses, and lipid peroxidation.

[Effects of different frequency electromagnetic fields on the differentiation of midbrain neural stem cells].

Objective. To examine the effect of electromagnetic fields (EMF) (20 Hz, 8 mT; 5 Hz, 8 mT) on the neuron-orientated differentiation of neural stem cells (NSCs) from midbrains of new-bom rats. Method. Differentiated NSCs were exposed to EMF for 2 x 15 min per day lasting for I d, 5 d, or 10 d. The sham-exposure controls were correspondingly established. Cells were fixed and processed for immunofluorescent staining using the antibody against neuron-specific marker MAP2, then the percentage of MAP2+ cells was calculated. Result. The two EMFs promoted the differentiation of a neuronal fate in different ways. Both of them came into effect even after 1 d exposure. When cells exposed to the 20 Hz EMF, the percentage of neuron-orientated cells gradually increased with longer-term exposure and the most significant effect appeared in 10 d group while that happened in 5 d group under the condition of 5 Hz EMF. The effect contrasted horizontally, significant differences between the two EMFs were observed only at 10 d groups, 20 Hz EMF having more favorable effect than 5 Hz EMF. Conclusion. 20 Hz and 5 Hz EMF could promote the differentiation of midbrain NSCs to a neuronal phenotype in different ways, suggesting that the physical induction might be another strategy to manipulate the differentiation of NSCs.

Spatially reciprocal inhibition of inhibition within a stimulus selection network in the avian midbrain.

Reciprocal inhibition between inhibitory projection neurons has been proposed as the most efficient circuit motif to achieve the flexible selection of one stimulus among competing alternatives. However, whether such a motif exists in networks that mediate selection is unclear. Here, we study the connectivity within the nucleus isthmi pars magnocellularis (Imc), a GABAergic nucleus that mediates competitive selection in the midbrain stimulus selection network. Using laser photostimulation of caged glutamate, we find that feedback inhibitory connectivity is global within the Imc. Unlike typical lateral inhibition in other circuits, intra-Imc inhibition remains functionally powerful over long distances. Anatomically, we observed long-range axonal projections and retrograde somatic labeling from focal injections of bi-directional tracers in the Imc, consistent with spatial reciprocity of intra-Imc inhibition. Together, the data indicate that spatially reciprocal inhibition of inhibition occurs throughout the Imc. Thus, the midbrain selection circuit possesses the most efficient circuit motif possible for fast, reliable, and flexible selection.

Effective dose span of ten different cone beam CT devices.

OBJECTIVES: Evaluation and reduction of dose are important issues. Since cone beam CT (CBCT) has been established now not just in dentistry, the number of acquired examinations continues to rise. Unfortunately, it is very difficult to compare the doses of available devices on the market owing to different exposition parameters, volumes and geometries. The aim of this study was to evaluate the spans of effective doses (EDs) of ten different CBCT devices. METHODS: 48 thermoluminescent dosemeters were placed in 24 sites in a RANDO(®) head phantom. Protocols with lowest exposition parameters and protocols with highest exposition parameters were performed for each of the ten devices. The ED was calculated from the measured energy doses according to the International Commission on Radiological Protection 2007 recommendations for each protocol and device, and the statistical values were evaluated afterwards. RESULTS: The calculation of the ED resulted in values between 17.2 µSv and 396 µSv for the ten devices. The mean values for protocols with lowest and highest exposition parameters were 31.6 µSv and 209 µSv, respectively. CONCLUSIONS: It was not the aim of this study to evaluate the image quality depending on different exposition parameters but to define the spans of EDs in which different CBCT devices work. There is a wide span of ED for different CBCT devices depending on the selected exposition parameters, required spatial resolution and many other factors.

Neuroprotection of midbrain dopaminergic cells in MPTP-treated mice after near-infrared light treatment.

This study explores whether near-infrared (NIr) light treatment neuroprotects dopaminergic cells in the substantia nigra pars compacta (SNc) and the zona incerta-hypothalamus (ZI-Hyp) from degeneration in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice. BALB/c albino mice were divided into four groups: 1) Saline, 2) Saline-NIr, 3) MPTP, 4) MPTP-NIr. The injections were intraperitoneal and they were followed immediately by NIr light treatment (or not). Two doses of MPTP, mild (50 mg/kg) and strong (100 mg/kg), were used. Mice were perfused transcardially with aldehyde fixative 6 days after their MPTP treatment. Brains were processed for tyrosine hydroxylase (TH) immunochemistry. The number of TH(+) cells was estimated using the optical fractionator method. Our major finding was that in the SNc there were significantly more dopaminergic cells in the MPTP-NIr compared to the MPTP group (35%-45%). By contrast, in the ZI-Hyp there was no significant difference in the numbers of cells in these two groups. In addition, our results indicated that survival in the two regions after MPTP insult was dose-dependent. In the stronger MPTP regime, the magnitude of loss was similar in the two regions ( approximately 60%), while in the milder regime cell loss was greater in the SNc (45%) than ZI-Hyp ( approximately 30%). In summary, our results indicate that NIr light treatment offers neuroprotection against MPTP toxicity for dopaminergic cells in the SNc, but not in the ZI-Hyp.