Studying the magnetic stimulation of nervous tissues: A calculation framework to investigate stimulation areas.

The electromagnetic stimulation of nervous tissue has represented an alternative to electrical stimulation since the 1980s. The growing number of potential applications has led to an increasing interest in the development of modeling tools that can help the design of novel electromagnetic stimulators. In this context, the aim of this paper is to provide a versatile calculation framework to investigate the properties of the electric field generated by a plurality of miniature coils, arranged in cuff configuration. Furthermore, the capability of the miniature coils to elicit a neuronal response in specific portions of the (peripheral) nerve will be investigated. Starting from Jefimenko's equations, a model was implemented in MATLAB. It calculates the electromagnetic field induced by coils, with arbitrary shape and spatial orientation, and the activating function around the coils through simple numerical integration. By studying the activating functions, it is possible to determine where the neurons can be excited. The model was validated by comparison with FEM simulations. A dimensional analysis was conducted to compare in terms of shape and depth of the stimulation volumes different coil geometries, regardless of design parameters such as current, number of turns and coil sizes.The dimensionless groups identified according to Buckingham's theorem provide a direct estimate of the stimulation depth reached within the nerve.The calculation tools developed in this paper can be used in the design of coils to quickly compare different geometries and spatial distribution of coils in order to identify the optimal configurations for the specific application addressed by the designer.

Evidence of cellular stress and caspase-3 resulting from a combined two-frequency signal in the cerebrum and cerebellum of sprague-dawley rats.

Multiple simultaneous exposures to electromagnetic signals induced adjustments in mammal nervous systems. In this study, we investigated the non-thermal SAR (Specific Absorption Rate) in the cerebral or cerebellar hemispheres of rats exposed in vivo to combined electromagnetic field (EMF) signals at 900 and 2450 MHz.Forty rats divided into four groups of 10 were individually exposed or not exposed to radiation in a GTEM chamber for one or two hours. After radiation, we used the Chemiluminescent Enzyme-Linked Immunosorbent Assay (ChELISA) technique to measure cellular stress levels, indicated by the presence of heat shock proteins (HSP) 90 and 70, as well as caspase-3-dependent pre-apoptotic activity in left and right cerebral and cerebellar hemispheres of Sprague Dawley rats.Twenty-four hours after exposure to combined or single radiation, significant differences were evident in HSP 90 and 70 but not in caspase 3 levels between the hemispheres of the cerebral cortex at high SAR levels. In the cerebellar hemispheres, groups exposed to a single radiofrequency (RF) and high SAR showed significant differences in HSP 90, 70 and caspase-3 levels compared to control animals. The absorbed energy and/or biological effects of combined signals were not additive, suggesting that multiple signals act on nervous tissue by a different mechanism.

[Therapeutic potential of optogenetic neuromodulation]. / Potentiel thérapeutique de la neuromodulation optogénétique.

Optogenetic neuromodulation techniques, which have emerged during the last 15 years, have considerably enhanced our ability to probe the functioning of neural circuits by allowing the excitation and inhibition of genetically-defined neuronal populations using light. Having gained tremendous popularity in the field of fundamental neuroscience, these techniques are now opening new therapeutic avenues. Optogenetic neuromodulation is a method of choice for studying the physiopathology of neurological and neuropsychiatric disorders in a range of animal models, and could accelerate the discovery of new therapeutic strategies. New therapeutic protocols employing optogenetic neuromodulation may also emerge in the near future, offering promising alternative approaches for disorders which lack appropriate treatments, such as pharmacoresistant epilepsy and inherited retinal degeneration.

Laser microbeam targeting of single nerve axons in cell culture.

By focusing a laser with short pulses to a diffraction-limited spot, single nerve axons can be precisely targeted and injured. Subsequent repair can be analyzed using various imaging and biochemical techniques to understand the repair process. In this chapter, we will describe a robotic laser microscope system used to injure nerve axons while simultaneously observing repair using phase and fluorescence microscopy. We provide procedures for controlled laser targeting and an experimental approach for studying axonal repair in embryonic rat hippocampus neurons.

Interdependence of Bad and Puma during ionizing-radiation-induced apoptosis.

Ionizing radiation (IR)-induced DNA double-strand breaks trigger an extensive cellular signaling response that involves the coordination of hundreds of proteins to regulate DNA repair, cell cycle arrest and apoptotic pathways. The cellular outcome often depends on the level of DNA damage as well as the particular cell type. Proliferating zebrafish embryonic neurons are highly sensitive to IR-induced apoptosis, and both p53 and its transcriptional target puma are essential mediators of the response. The BH3-only protein Puma has previously been reported to activate mitochondrial apoptosis through direct interaction with the pro-apoptotic Bcl-2 family proteins Bax and Bak, thus constituting the role of an "activator" BH3-only protein. This distinguishes it from BH3-only proteins like Bad that are thought to indirectly promote apoptosis through binding to anti-apoptotic Bcl-2 family members, thereby preventing the sequestration of activator BH3-only proteins and allowing them to directly interact with and activate Bax and Bak. We have shown previously that overexpression of the BH3-only protein Bad in zebrafish embryos supports normal embryonic development but greatly sensitizes developing neurons to IR-induced apoptosis. While Bad has previously been shown to play only a minor role in promoting IR-induced apoptosis of T cells in mice, we demonstrate that Bad is essential for robust IR-induced apoptosis in zebrafish embryonic neural tissue. Moreover, we found that both p53 and Puma are required for Bad-mediated radiosensitization in vivo. Our findings show the existence of a hierarchical interdependence between Bad and Puma whereby Bad functions as an essential sensitizer and Puma as an essential activator of IR-induced mitochondrial apoptosis specifically in embryonic neural tissue.

Radiation impairs perineural invasion by modulating the nerve microenvironment.

PURPOSE: Perineural invasion (PNI) by cancer cells is an ominous clinical event that is associated with increased local recurrence and poor prognosis. Although radiation therapy (RT) may be delivered along the course of an invaded nerve, the mechanisms through which radiation may potentially control PNI remain undefined. EXPERIMENTAL DESIGN: An in vitro co-culture system of dorsal root ganglia (DRG) and pancreatic cancer cells was used as a model of PNI. An in vivo murine sciatic nerve model was used to study how RT to nerve or cancer affects nerve invasion by cancer. RESULTS: Cancer cell invasion of the DRG was partially dependent on DRG secretion of glial-derived neurotrophic factor (GDNF). A single 4 Gy dose of radiation to the DRG alone, cultured with non-radiated cancer cells, significantly inhibited PNI and was associated with decreased GDNF secretion but intact DRG viability. Radiation of cancer cells alone, co-cultured with non-radiated nerves, inhibited PNI through predominantly compromised cancer cell viability. In a murine model of PNI, a single 8 Gy dose of radiation to the sciatic nerve prior to implantation of non-radiated cancer cells resulted in decreased GDNF expression, decreased PNI by imaging and histology, and preservation of sciatic nerve motor function. CONCLUSIONS: Radiation may impair PNI through not only direct effects on cancer cell viability, but also an independent interruption of paracrine mechanisms underlying PNI. RT modulation of the nerve microenvironment may decrease PNI, and hold significant therapeutic implications for RT dosing and field design for patients with cancers exhibiting PNI.

[Er:YAG laser: tissue interaction and histomorphological characterization]. / Laser a Erbium YAG: caratteristiche istomorfologiche da interazione energia-tessuto.

Authors describe Er:YAG laser interaction with tissues, in particular their histomorphological characteristics to identify a specific clinic area for laser application through the examination of different clinical international trials. This study includes experimental trials about pig and rat skin laser application to know laser Er: YAG capability and limits; investigation is extended to laser application in human soft tissues as mucosa, periosteum and bones, its utility in cutaneous pathologies and in antiageing treatments.

Combined optical and electrical stimulation of neural tissue in vivo.

Low-intensity, pulsed infrared light provides a novel nerve stimulation modality that avoids the limitations of traditional electrical methods such as necessity of contact, presence of a stimulation artifact, and relatively poor spatial precision. Infrared neural stimulation (INS) is, however, limited by a 2:1 ratio of threshold radiant exposures for damage to that for stimulation. We have shown that this ratio is increased to nearly 6:1 by combining the infrared pulse with a subthreshold electrical stimulus. Our results indicate a nonlinear relationship between the subthreshold depolarizing electrical stimulus and additional optical energy required to reach stimulation threshold. The change in optical threshold decreases linearly as the delay between the electrical and optical pulses is increased. We have shown that the high spatial precision of INS is maintained for this combined stimulation modality. Results of this study will facilitate the development of applications for infrared neural stimulation, as well as target the efforts to uncover the mechanism by which infrared light activates neural tissue.

Expression of the melatonin receptor Mel(1c) in neural tissues of the reef fish Siganus guttatus.

The golden rabbitfish, Siganus guttatus, is a reef fish exhibiting a restricted lunar-related rhythm in behavior and reproduction. Here, to understand the circadian rhythm of this lunar-synchronized spawner, a melatonin receptor subtype-Mel(1c)-was cloned. The full-length Mel(1c) melatonin receptor cDNA comprised 1747 bp with a single open reading frame (1062 bp) that encodes a 353-amino acid protein, which included 7 presumed transmembrane domains. Real-time PCR revealed high Mel(1c) mRNA expression in the retina and brain but not in the peripheral tissues. When the fish were reared under light/dark (LD 12:12) conditions, Mel(1c) mRNA in the retina and brain was expressed with daily variations and increased during nighttime. Similar variations were noted under constant conditions, suggesting that Mel(1c) mRNA expression is regulated by the circadian clock system. Daily variations of Mel(1c) mRNA expression with a peak at zeitgeber time (ZT) 12 were observed in the cultured pineal gland under LD 12:12. Exposure of the cultured pineal gland to light at ZT17 resulted in a decrease in Mel(1c) mRNA expression. When light was obstructed at ZT5, the opposite effect was obtained. These results suggest that light exerts certain effects on Mel(1c) mRNA expression directly or indirectly through melatonin actions.

Simultaneous birefringence and scattered light measurements reveal anatomical features in isolated crustacean nerve.

Simultaneous fast birefringence and scattered light changes associated with crustacean nerve activation have different time courses and are produced by separate biophysical mechanisms. Technological advances in illumination, photodiodes and amplification circuitry achieved better signal-to-noise than earlier studies revealing optical signals in axonal nerve bundles as small as crayfish ventral cord and claw. The birefringence measurements yielded signals that could be observed in single trials, with temporally separated peaks associated with axonal populations of different diameters. A slit aperture placed perpendicular to the nerve reduced the spatial-temporal integration and enhanced the temporal structure of the separate peaks in the birefringence signal. Moving the slit aperture farther from the stimulation point delayed the signal in time, and also enhanced the separation between peaks. Different propagation velocities of the separate peaks provided evidence for at least three neuronal populations in the bundle. These studies underscore the advantages of birefringence over scattering measurements. Application of birefringence methods can optimize non-invasive imaging techniques being developed to detect fast optical responses associated with electrical neural activity in humans.

[The effect of the x-ray irradiation of rats on the NAD-pyrophosphorylase and poly(ADP-ribose)polymerase activities of brain nuclei and on the NAD content in nerve tissue]. / Vliianie rentgenovskogo oblucheniia krys na NAD-pirofosforilaznuiu i poli(ADP-riboza)polimeraznuiu aktivnost' iader golovnogo mozga i na soderzhanie NAD v nervnoi tkani.

A study was made of the influence of X-irradiation of rats with various doses on NAD-pyrophosphorylase and poly(ADP-ribose) polymerase activity of brain nuclei. It was shown that X-radiation was ineffective with regard to NAD-pyrophosphorylase activity of nuclei and increased their poly(ADP-ribose) polymerase activity. Stimulation of poly(ADP-ribose) polymerase activity of nuclei was a function of radiation dose and correlated with the decrease in the NAD content of nervous tissue. It was found that mainly nonhistone proteins were ADP-ribosylated in nuclei of both irradiated and nonirradiated rats.

Distribution of phosphate-independent MAP2 epitopes revealed with monoclonal antibodies in microwave-denatured human nervous system tissues.

In contrast with results obtained in experimental animals, antibodies to microtubule associated protein-2 (MAP2) preferentially label abnormal structures in human nervous system tissue samples, but the normal sites at which MAP2 is expressed are not well-defined. To determine the distribution of MAP2 in the human central (CNS) and peripheral (PNS) nervous systems, we prepared monoclonal antibodies (MAbs) specific to MAP2, and compared the localization of this MAP in postmortem bovine and human tissues as well as in several human neural cell lines that express either neurofilament (NF) or glial filament (GF) proteins. Eight MAbs specific for phosphate-independent epitopes in bovine and human MAP2 were obtained, and those that performed well in tissues produced immunoreactivity confined to the somatodendritic domain of neurons in bovine and human CNS and PNS tissues. Other neural cells (e.g. astrocytes) did not express MAP2 immunoreactivity using these MAbs. Postmortem delays of less than 24 h prior to tissue denaturation did not affect the distribution of MAP2 immunoreactivity. However, microwave denaturation of these tissues preserved MAP2 immunoreactivity better than fixation with Bouin's solution or formalin. Microwave treatment also improved the immunoreactivity of several MAbs for NF and GF proteins. Finally, MAP2 was not detected in human neural cell lines that express NF (2) or GF (1) proteins. We conclude that microwave denaturation provides an effective means to preserve the immunoreactivity of normal human neuronal cytoskeletal proteins, and that this method of tissue denaturation allows the normal distribution of MAP2 to be defined in postmortem samples of human CNS and PNS tissues.

Contractile properties of standard and nerve-intact muscle grafts in the rat.

The contractile properties of standard and nerve-intact grafts of extensor digitorum longus muscles of rats were compared in vitro. Fourteen days after grafting, the time to peak tension and the half-relaxation times of nerve-intact grafts were shorter than those of standard grafts, but both were longer than control values. By 60 days, these variables attained normal values. At every sample period, the tetanic tensions of nerve-intact grafts were higher than those of standard grafts. Even at the early sampling periods, the twitch-tetanic tension ratios of nerve-intact grafts were close to normal values, whereas those of standard grafts were higher than normal. Stabilized nerve-intact grafts had a larger mass and greater maximum tetanic tension development than standard grafts, but were more fatigable. Compared with control EDL muscles, stable nerve-intact grafts show no differences except for lessened fatigability, whereas standard grafts demonstrate significant functional deficits.

The subcellular basis of damage to the human urinary bladder induced by irradiation.

The effects of x-irradiation on the subcellular structure of the human urinary bladder were investigated by electron microscopic examination of biopsies taken during check cystoscopies from 25 patients between 1 month and 22 years after completion of a course of therapeutic radiation. All tissues of the bladder wall were damaged to some extent by the treatment. In the urothelium this was reflected by the development of more than the usual numbers of lysosomes and autophagic vacuoles in all cell layers. In the bladder wall, large often binucleate or multinucleate fibroblasts were prominent and persistent in all specimens and were associated with the development of progressive fibrosis. The vasculature and the muscle coats of the bladder wall were also damaged. In the blood vessels many endothelial cells were oedematous or necrotic and some intravascular coagulation was also observed. Smooth muscle cells became oedematous soon after irradiation, and after longer time intervals there was focal death and loss of individual muscle cells. The observed degeneration and extensive necrosis of the bladder wall, which involved severe destruction and disorganization of the muscular layers, is sufficient to explain the clinical sequelae of bladder irradiation, namely loss of elasticity, reduced capacity and incomplete micturition with residual urine.

The laser in neurological surgery.

The use of lasers in neurosurgical procedures has received a great deal of attention recently. Surgical use of lasers has been viewed with suspicion and skepticism, probably because of (justified) apprehensions about the misuse of lasers in early work and about the ways in which laser light affects tissues, and a lack of understanding of the basic physics and practical operation of lasers. The authors review the physics, biophysics, experimental findings, and operative use of lasers in current neurosurgical practice, and discuss briefly their experience gained in over 150 neurosurgical procedures using the carbon dioxide and argon surgical lasers.

Photosensitization and the nervous system in the planarian Dugesia gonocephala. A histochemical, ultrastructural and behavioral investigation.

Photosensitization phenomena may be induced in planarias by eosin and hematoporphyrin, and as a result, dopamine agonistic behavior ("screw-like hyperkinesia") is set up in the animal. Histochemical, ultrastructural and pharmacological investigations have shown that this hyperkinesia is of post-synaptic origin in eosin photosensitization, and of pre-synaptic origin in hematoporphyrin photosensitization. The authors suggest an hypothesis to explain the different activity of the two photosensitizers, and discuss the validity of the experiment with regard to human porphyria.