Axonal Accumulation of Lysosomal-Associated Membrane Protein 1 (LAMP1) Accompanying Alterations of Autophagy Dynamics in the Rat Hippocampus Upon Seizure-Induced Injury.

We found a dramatic upregulation in the expression of LC3 in the hippocampus of rats upon status epilepticus (SE). However, the enhancement in LC3 expression might be caused by a reduction in lysosomal activity or by alterations in autophagosome-lysosome fusion leading to a cytosolic vesicular retention. In order to dissect this aspect, we monitored the spatial and temporal expression of LC3 and LAMP1 in the hippocampus of rats with SE. The Western blot analysis showed that the expression of LAMP1 was slightly increased in hippocampal cells at 6, 24, and 48 h post-SE. However, immunofluorescence analysis showed dramatic spatial changes in LAMP1 distribution within the hippocampus. LAMP1 in controls was localised only in cytosol as dot like staining, however at 24 h post-SE LAMP1 was not only highly expressed, but accumulated in mossy fibers of dentate gyrus. In parallel, we found few scattered LC3-positive-dots in neurites of dentate gyrus which co-localise with LAMP1-positive structures. We conclude that SE not only increased autophagosomal abundance, but also lysosomal activities and a massive accumulation of LAMP1 in axons of dentate gyrus. This could support the hypothesis that the marked increased autophagosomal abundance in cytosol reflects an increase in the autophagic activity more than an inhibition of autophagosomal clearance. Although LAMP1 may have contributed to cell damage in the selective vulnerable hippocampal CA1-subfield, it is also possible that lysosomal/autophagic mechanisms in mossy fibers were compensatory and reflected an attempt to survive the epileptic insult by breaking down non-essential components.

Monolithic metallic nanocavities for strong light-matter interaction to quantum-well intersubband excitations.

We present the design, realization and characterization of strong coupling between an intersubband transition and a monolithic metamaterial nanocavity in the mid-infrared spectral range. We use a ground plane in conjunction with a planar metamaterial resonator for full three-dimensional confinement of the optical mode. This reduces the mode volume by a factor of 1.9 compared to a conventional metamaterial resonator while maintaining the same Rabi frequency. The conductive ground plane is implemented using a highly doped n+ layer which allows us to integrate it monolithically into the device and simplify fabrication.

THz quantum cascade lasers with wafer bonded active regions.

We demonstrate terahertz quantum-cascade lasers with a 30 µm thick double-metal waveguide, which are fabricated by stacking two 15 µm thick active regions using a wafer bonding process. By increasing the active region thickness more optical power is generated inside the cavity, the waveguide losses are decreased and the far-field is improved due to a larger facet aperture. In this way the output power is increased by significantly more than a factor of 2 without reducing the maximum operating temperature and without increasing the threshold current.

[Significance of frozen section diagnosis for the management of laryngeal tumors]. / Bedeutung der Schnellschnittdiagnostik für das Management laryngealer Tumoren.

The frozen section procedure for immediate intraoperative pathological diagnosis represents a pivotal method in tumor diagnosis. In laryngeal tumors the most frequent indication for the use of this method is the documentation of the residual tumor status, while intraoperative consultation with the purpose of primary tumor diagnosis is less common. The specimen management employed in each case should be chosen depending on the clinical question: while the collection of a maximum amount of tissue is advisable for the determination of the residual tumor status, sparing a portion of the remaining tissue for possible future examinations is advisable in the case of primary tumor diagnosis. Moreover, intraoperative frozen section diagnosis with no immediate consequences should be avoided.

Terahertz meta-atoms coupled to a quantum well intersubband transition.

We present a method of coupling free-space terahertz radiation to intersubband transitions in semiconductor quantum wells using an array of meta-atoms. Owing to the resonant nature of the interaction between metamaterial and incident light and the field enhancement in the vicinity of the metal structure, the coupling efficiency of this method is very high and the energy conversion ratio from in-plane to z field reaches values on the order of 50%. To identify the role of different aspects of this coupling, we have used a custom-made finite-difference time-domain code. The simulation results are supplemented by transmission measurements on modulation-doped GaAs/AlGaAs parabolic quantum wells which demonstrate efficient strong light-matter coupling between meta-atoms and intersubband transitions for normal incident electromagnetic waves.

Time- and momentum-resolved photoemission studies using time-of-flight momentum microscopy at a free-electron laser.

Time-resolved photoemission with ultrafast pump and probe pulses is an emerging technique with wide application potential. Real-time recording of nonequilibrium electronic processes, transient states in chemical reactions, or the interplay of electronic and structural dynamics offers fascinating opportunities for future research. Combining valence-band and core-level spectroscopy with photoelectron diffraction for electronic, chemical, and structural analyses requires few 10 fs soft X-ray pulses with some 10 meV spectral resolution, which are currently available at high repetition rate free-electron lasers. We have constructed and optimized a versatile setup commissioned at FLASH/PG2 that combines free-electron laser capabilities together with a multidimensional recording scheme for photoemission studies. We use a full-field imaging momentum microscope with time-of-flight energy recording as the detector for mapping of 3D band structures in (kx, ky, E) parameter space with unprecedented efficiency. Our instrument can image full surface Brillouin zones with up to 7 Å-1 diameter in a binding-energy range of several eV, resolving about 2.5 × 105 data voxels simultaneously. Using the ultrafast excited state dynamics in the van der Waals semiconductor WSe2 measured at photon energies of 36.5 eV and 109.5 eV, we demonstrate an experimental energy resolution of 130 meV, a momentum resolution of 0.06 Å-1, and a system response function of 150 fs.

Active photonic crystal terahertz laser.

We present the design and the realization of active photonic crystal (PhC) semiconductor lasers. The PhC consists of semiconductor nanostructure pillars which provide gain at a quantized transition energy. The vertical layer sequence is that of a terahertz quantum cascade laser. Thereby, the artificial crystal itself provides the optical gain and the lateral confinement. The cavities do not rely on a central defect, the lasing is observed in flat-band regions at high symmetry points. The experimental results are in excellent agreement with the finite-difference time-domain simulations. For the vertical confinement a double-metal waveguide is used. The lasers are showing a stable single-mode emission under all driving conditions. Varying the period of the PhC allows to tune the frequency by 400 GHz.

Electrically controllable photonic molecule laser.

We have studied the coherent intercavity coupling of the evanescent fields of two microdisk terahertz quantum-cascade lasers. The electrically controllable optical coupling of the single-mode operating lasers has been observed for cavity spacings up to 30 mum. The strongest coupled photonic molecule with 2 mum intercavity spacing allows to conditionally switch the optical emission by the electrical modulation of only one microdisk. The lasing threshold characteristics demonstrate the linear dependence of the gain of a quantum-cascade laser on the applied electric field.

Discovery of microwave emission from four nearby solar-type g stars.

Radio waves from the sun were detected 50 years ago, but the microwave detection of other single solar-type stars has remained a challenge. Here, the discovery of four solar-type radio stars is reported. These "solar twin" G stars are radio sources up to 3000 times stronger than the quiet sun. The microwaves most likely originate from a large number of relativistic electrons, possibly produced along with coronal heating, a process that is not understood. Two of the stars are younger than the sun and rotate more rapidly; the dynamo process in the stellar interior is therefore presumably more vigorous, resulting in enhanced coronal activity. One of the detections, however, is an old, metal-deficient G dwarf.

Exclusive Activation of Caspase-3 in Mossy Fibers and Altered Dynamics of Autophagy Markers in the Mice Hippocampus upon Status Epilepticus Induced by Kainic Acid.

Epileptic seizures are generally associated with pathological changes in the hippocampus such as astrogliosis, mossy fiber sprouting, and neuronal damage. However, more than 30% of temporal lobe epilepsy in humans shows neither neuronal damage nor mossy fiber sprouting despite chronic epileptic seizures. A similar situation exists in certain commonly used strains of mice, specifically C57BL/6 and BALB/c, which exhibit epileptic seizures, but no neuronal damage upon kainic acid administration. This suggests that intrinsic factors may influence the pathological manifestations of epilepsy. Mechanisms which are behind the resistance of hippocampal cells to KA-induced neuronal death are unknown. Autophagy seems to be involved in the pathogenesis of many brain insults and to have a dual nature in neuroprotection and cell death. This study addresses the role of autophagy upon status epilepticus (SE) that has been induced by kainic acid (KA) in the C57BL/6 strain which is classified as seizure resistant. We analyzed the dynamics in the expression of autophagic and cell death markers in the hippocampus upon SE. Immunofluorescence data show that KA did not induce neuronal death in the hippocampal CA1-CA3 subfields; however, it leads to an exclusive activation of caspase-3 in the mossy fibers. We also found alterations in the expression of core proteins of the autophagic machinery. Levels of MAP1LC3, phospho-mTOR/mTOR, and Beclin 1 were significantly increased after induction of seizures. However, levels of Atg3, Atg14, Atg5-Atg12, Atg7, BAG3, Hsp70, and LAMP1 showed no significant alterations compared to controls. Although KA did not induce neuronal death, this study provides morphological and biochemical evidence that status epilepticus induced by KA activates caspase-3 in mossy fibers and induces autophagy in the C57BL/6 hippocampus. These data indicate that autophagic factors may modulate the sensitivity of pyramidal cells to KA and that autophagy may constitute a part of an endogenous neuroprotective arsenal which might be behind the resistance of C57BL/6-hippocampal cells to KA-induced neuronal death.

beta-Hydroxybutyrate fuels synaptic function during development. Histological and physiological evidence in rat hippocampal slices.

To determine whether ketone bodies sustain neuronal function as energy substrates, we examined the effects of beta-hydroxybutyrate (betaHB) on synaptic transmission and morphological integrity during glucose deprivation in rat hippocampal slices. After the depression of excitatory postsynaptic potentials (EPSPs) by 60 min of glucose deprivation, administration of 0.5-10 mM D-betaHB restored EPSPs in slices from postnatal day (PND) 15 rats but not in slices from PND 30 or 120 rats. At PND 15, adding D-betaHB to the media allowed robust long-term potentiation of EPSPs triggered by high frequency stimulation, and prevented the EPSP-spike facilitation that suggests hyperexcitability of neurons. Even after PND 15,D-betaHB blocked morphological changes produced by either glucose deprivation or glycolytic inhibition. These results indicate that D-betaHB is not only able to substitute for glucose as an energy substrate but is also able to preserve neuronal integrity and stability, particularly during early development.

Terahertz photonic crystal resonators in double-metal waveguides.

We present the design and the fabrication of photonic crystals with a complete bandgap for TM-modes used as a resonator for terahertz quantum-cascade lasers (QCL), which are lasing around 2.7 THz. The emission of the devices with and without a photonic crystal shows a shift in the emission from the gain maximum to the bandgap of the crystal. The devices are built up by a core, which provides the optical gain, and by a surrounding photonic crystal, which acts as a frequency selective mirror. The whole device is processed into a double-metal waveguide.

Expression of fructose sensitive glucose transporter in the brains of fructose-fed rats.

Glucose transporters play a critical role in mammalian brain energy metabolism because glucose is the principal brain energy source and these transporters promote glucose movement into neural cells. When glucose is unavailable, fructose can serve as an alternative energy source. Using real-time polymerase chain reaction and actin as a reference mRNA, we investigated the impact of fructose feeding on rat brain and other tissue mRNA expression of glucose transporter 5 which has high affinity for fructose. Brain mRNA levels of glucose transporter 5 increased 1.5-fold in 35-day old rats after 7 days of fructose feeding compared with controls, whereas it increased 2.5-fold in jejunum. Semi-quantitative analysis of protein expression by immunofluorescence of glucose transporter 5 in rat hippocampi indicated a 2.4-fold increase. We demonstrated the specificity of fructose feeding on glucose transporter 5 expression by showing that the expression of the neuronal glucose transporter 3 and insulin-regulated glucose transporter 4 were unaffected. In addition, the expression of glucose transporter 5 increased in fructose fed older adult rats (8-months and 12-months old) when compared with controls. These results suggest that short-term fructose feeding increases the expression of glucose transporter 5 in both young and aging adult rats. Increased brain expression of glucose transporter 5 is likely to be important in the role of fructose as an alternative energy source.

Substrate turnover by transporters curtails synaptic glutamate transients.

Although inhibitors of glutamate transport prolong synaptic currents at many glutamate synapses, the cause of the current prolongation is unclear. Transport inhibitors may prolong synaptic currents by simply interfering with synaptic glutamate binding to transporters, by inhibiting substrate translocation, or by promoting accumulation of ambient glutamate, which may act cooperatively at receptors with synaptic glutamate. We show that reversal of the membrane potential of astrocytes surrounding the synapse prolongs synaptic currents but does not decrease the apparent affinity of transporters or significantly alter glutamate-dependent kinetics of macroscopic transporter currents in excised membrane patches. Positive membrane potentials do not affect binding of a nontransported glutamate analog, nor do positive membrane potentials alter the number of transporters available to bind analog. We also test the hypothesis that glutamate accumulation during uptake inhibition by transporter substrates is the direct cause of synaptic current prolongations. Transporter substrates elevate ambient glutamate near synapses by fostering reverse transport of endogenous glutamate. However, increases in ambient glutamate cannot account for the prolongations of synaptic currents, because a nonsubstrate transport inhibitor does not foster reverse uptake yet it prolongs synaptic currents. Moreover, exogenous glutamate does not mimic synaptic current prolongations induced by substrate inhibitors. These results provide strong support for a major role of substrate translocation in determining the time course of the glutamate concentration transient at excitatory synapses.

Astrocytes exert a pro-apoptotic effect on neurons in postnatal hippocampal cultures.

We investigated conditions that promote basal and activity-dependent neuronal apoptosis in postnatal rat hippocampal cultures. Low-density mixed cultures of astrocytes and neurons exhibited lower sensitivity than high-density cultures to basal neuronal death and activity-sensitive neuronal death, induced with glutamate receptor blockers, sodium channel blockers, or calcium channel blockers. Although elevations of [Ca(2+)](i) protect neurons from apoptosis, low-density microcultures and mass cultures exhibited only minor differences in resting [Ca(2+)](i) and Ca(2+) current density, suggesting that these variables are unlikely to explain differences in susceptibility. Astrocytes, rather than neurons, were implicated in the neuronal loss. Several candidate molecules implicated in other astrocyte-dependent neurotoxicity models were excluded, but heat inactivation experiments suggested that a heat-labile factor is critically involved. In sum, our results suggest the surprising result that astrocytes can be negative modulators of neuronal survival during development and when the immature nervous system is challenged with drugs that dampen electrical excitability.

Oxygen deprivation produces delayed inhibition of long-term potentiation by activation of NMDA receptors and nitric oxide synthase.

The acute and delayed effects of anoxia on synaptic transmission and long-term potentiation (LTP) were examined in the CA1 region of rat hippocampal slices. Oxygen deprivation for 20 minutes completely but reversibly depressed excitatory postsynaptic potentials mediated by both N-methyl-D-aspartate receptors (NMDAR) and non-NMDAR. Although LTP was reliably produced by a single tetanus delivered 30 minutes after reoxygenation, LTP could not be induced when a tetanus was delivered 70 to 100 minutes after reoxygenation. A tetanus delivered 100 minutes after reoxygenation produced lasting synaptic enhancement when 100 mumol/L D,L-amino-phosphonovaleric acid (APV), a competitive NMDAR antagonist, was administered during the period of oxygen deprivation. The delayed effects of oxygen deprivation were not blocked when APV was administered after oxygen deprivation. Similarly, the delayed effects on LTP induction were overcome by inhibitors of nitric oxide synthase when the nitric oxide synthase inhibitors were administered during anoxia, but not when administered after oxygen deprivation. These results suggest that untimely activation of NMDAR and nitric oxide release during anoxia produce delayed inhibition of LTP induction and may be involved in the memory defects that occur subsequent to cerebral hypoxia.

Neuroprotective agent riluzole potentiates postsynaptic GABA(A) receptor function.

The antiepileptic drug riluzole is a use-dependent blocker of voltage-gated Na(+) channels and selectively depresses action potential-driven glutamate over gamma-aminobutyric acid (GABA) release. Here we report that in addition to its presynaptic effect, riluzole at higher concentrations also strongly potentiates postsynaptic GABA(A) responses both in cultured hippocampal neurons and in Xenopus oocytes expressing recombinant receptors. Although peak inhibitory postsynaptic currents (IPSCs) of autaptic hippocampal neurons were inhibited, 20-100 microM riluzole significantly prolonged the decay of IPSCs, resulting in little change in total charge transfer. The effect was dose-dependent and reversible. Riluzole selectively increased miniature IPSC fast and slow decay time constants, without affecting their relative proportions. Miniature IPSC peak amplitude, rise time and frequency were unaffected, indicating a postsynaptic mechanism. In the Xenopus oocyte expression system, riluzole potentiated GABA responses by lowering the EC(50) for GABA activation. Riluzole directly gated a GABA(A) current that was partially blocked by bicuculline and gabazine. Pharmacological experiments suggest that the action of riluzole did not involve a benzodiazepine, barbiturate, or neurosteroid site. Instead, riluzole-induced potentiation was inhibited by the lactone antagonist alpha-isopropyl-alpha-methyl-gamma-butyrolatone (alpha-IMGBL). While most anticonvulsants either block voltage-gated Na(+) channels or potentiate GABA(A) receptors, our results suggest that riluzole may define an advantageous class of anticonvulsants with both effects.

Effects of fructose-1,6-bisphosphate on morphological and functional neuronal integrity in rat hippocampal slices during energy deprivation.

D-fructose-1,6-bisphosphate, a high energy glycolytic intermediate, attenuates ischemic damage in a variety of tissues, including brain. To determine whether D-fructose-1,6-bisphosphate serves as an alternate energy substrate in the CNS, rat hippocampal slices were treated with D-fructose-1,6-bisphosphate during glucose deprivation. Unlike pyruvate, an endproduct of glycolysis, 10 mM D-fructose-1,6-bisphosphate did not preserve synaptic transmission or morphological integrity of CA1 pyramidal neurons during glucose deprivation. Moreover, during glucose deprivation, 10-mM D-fructose-1,6-bisphosphate failed to maintain adenosine triphosphate levels in slices. D-fructose-1,6-bisphosphate, however, attenuated acute neuronal degeneration produced by 200 microM iodoacetate, an inhibitor of glycolysis downstream of D-fructose-1,6-bisphosphate. Because (5S, 10R)-(+)-5-methyl-10, 11-dihydro-5H-dibenzo [a,d]cyclohepten-5,10-imine, an antagonist of N-methyl-D-aspartate receptors, exhibited similar protection against iodoacetate damage, we examined whether (5S, 10R)-(+)-5-methyl-10, 11-dihydro-5H-dibenzo [a,d]cyclohepten-5,10-imine and D-fructose-1,6-bisphosphate share a common neuroprotective mechanism. Indeed, D-fructose-1,6-bisphosphate diminished N-methyl-D-aspartate receptor-mediated synaptic responses and partially attenuated neuronal degeneration induced by 100-microM N-methyl-D-aspartate. Taken together, these results indicate that D-fructose-1,6-bisphosphate is unlikely to serve as an energy substrate in the hippocampus, and that neuroprotective effects of D-fructose-1,6-bisphosphate are mediated by mechanisms other than anaerobic energy supply.

Effects of lactate and pyruvate on glucose deprivation in rat hippocampal slices.

Rat hippocampal slices were used to evaluate the effects of glucose deprivation and the ability of lactate or pyruvate to preserve histological integrity and synaptic function. Dark cell changes were observed during 180 min incubations in glucose-free solutions. These changes were blocked by substituting 10 mM lactate or pyruvate for glucose during the incubation. Excitatory postsynaptic potentials disappeared during 60 min of glucose deprivation but were restored by subsequent introduction of glucose, lactate or pyruvate. Incubation of slices with iodoacetate revealed a distinct pattern of damage that was blocked completely by pyruvate and partially by lactate. These results indicate that exogenous pyruvate and lactate can serve as energy substrates in the hippocampus when glucose is unavailable or glycolytic metabolism is impaired.

Lactate dehydrogenase release is facilitated by brief sonication of rat hippocampal slices and isolated retinas following acute neuronal damage.

Although useful for determining neuronal damage in cell cultures, the lactate dehydrogenase (LDH) assay is not suitable for acute brain preparations because LDH release is typically delayed relative to neuronal deterioration. The slow release suggests that LDH may remain trapped inside damaged cells until late in the degenerative process. To test this, we examined whether brief sonication facilitates LDH release from acutely damaged neurons. In rat isolated retinas and hippocampal slices, LDH release was minimal following acute administration of iodoacetate or kainate. However, these toxins promoted significant LDH release, when toxin exposure was followed by brief sonication. Increases in extracellular LDH correlated with changes in neuronal morphology. These findings suggest that sonication may facilitate the use of the LDH assay in acute brain preparations.