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The appropriate duration of dual antiplatelet therapy in patients at high risk for bleeding after the implantation of a drug-eluting coronary stent remains unclear.One month after they had undergone implantation of a biodegradable-polymer sirolimus-eluting coronary stent, we randomly assigned patients at high bleeding risk to discontinue dual antiplatelet therapy immediately (abbreviated therapy) or to continue it for at least 2 additional months (standard therapy). The three ranked primary outcomes were net adverse clinical events (a composite of death from any cause, myocardial infarction, stroke, or major bleeding), major adverse cardiac or cerebral events (a composite of death from any cause, myocardial infarction, or stroke), and major or clinically relevant nonmajor bleeding; cumulative incidences were assessed at 335 days. The first two outcomes were assessed for noninferiority in the per-protocol population, and the third outcome for superiority in the intention-to-treat population.Among the 4434 patients in the per-protocol population, net adverse clinical events occurred in 165 patients (7.5%) in the abbreviated-therapy group and in 172 (7.7%) in the standard-therapy group (difference, -0.23 percentage points; 95% confidence interval [CI], -1.80 to 1.33; P0.001 for noninferiority). A total of 133 patients (6.1%) in the abbreviated-therapy group and 132 patients (5.9%) in the standard-therapy group had a major adverse cardiac or cerebral event (difference, 0.11 percentage points; 95% CI, -1.29 to 1.51; P = 0.001 for noninferiority). Among the 4579 patients in the intention-to-treat population, major or clinically relevant nonmajor bleeding occurred in 148 patients (6.5%) in the abbreviated-therapy group and in 211 (9.4%) in the standard-therapy group (difference, -2.82 percentage points; 95% CI, -4.40 to -1.24; P0.001 for superiority).One month of dual antiplatelet therapy was noninferior to the continuation of therapy for at least 2 additional months with regard to the occurrence of net adverse clinical events and major adverse cardiac or cerebral events; abbreviated therapy also resulted in a lower incidence of major or clinically relevant nonmajor bleeding. (Funded by Terumo; MASTER DAPT ClinicalTrials.gov number, NCT03023020.).

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cell carcinoma. Currently, there is a lack of noninvasive methods to stratify ccRCC prognosis prior to any invasive therapies. The purpose of this study was to preoperatively predict the tumor stage, size, grade, and necrosis (SSIGN) score of ccRCC using MRI-based radiomics.A multicenter cohort of 364 histopathologically confirmed ccRCC patients (272 low [ 4] and 92 high [≥ 4] SSIGN score) with preoperative T2-weighted and T1-contrast-enhanced MRI were retrospectively identified and divided into training (254 patients) and testing sets (110 patients). The performance of a manually optimized radiomics model was assessed by measuring accuracy, sensitivity, specificity, area under receiver operating characteristic curve (AUROC), and area under precision-recall curve (AUPRC) on an independent test set, which was not included in model training. Lastly, its performance was compared to that of a machine learning pipeline, Tree-Based Pipeline Optimization Tool (TPOT).The manually optimized radiomics model using Random Forest classification and Analysis of Variance feature selection methods achieved an AUROC of 0.89, AUPRC of 0.81, accuracy of 0.89 (95% CI 0.816-0.937), specificity of 0.95 (95% CI 0.875-0.984), and sensitivity of 0.72 (95% CI 0.537-0.852) on the test set. The TPOT using Extra Trees Classifier achieved an AUROC of 0.94, AUPRC of 0.83, accuracy of 0.89 (95% CI 0.816-0.937), specificity of 0.95 (95% CI 0.875-0.984), and sensitivity of 0.72 (95% CI 0.537-0.852) on the test set.Preoperative MR radiomics can accurately predict SSIGN score of ccRCC, suggesting its promise as a prognostic tool that can be used in conjunction with diagnostic markers.

Stellar mass has been shown to correlate with halo mass, with non-negligible scatter. The stellar mass-size and luminosity-size relationships of galaxies also show significant scatter in galaxy size at fixed stellar mass. It is possible that, at fixed stellar mass and galaxy colour, the halo mass is correlated with galaxy size. Galaxy-galaxy lensing allows us to measure the mean masses of dark matter haloes for stacked samples of galaxies. We extend the analysis of the galaxies in the CFHTLenS catalogue by fitting single S\'{e}rsic surface brightness profiles to the lens galaxies in order to recover half-light radius values, allowing us to determine halo masses for lenses according to their size. Comparing our halo masses and sizes to baselines for that stellar mass yields a differential measurement of the halo mass-galaxy size relationship at fixed stellar mass, defined as $M_{h}(M_{*}) \propto r_{\mathrm{eff}}^{\eta}(M_{*})$. We find that on average, our lens galaxies have an $\eta = 0.42\pm0.12$, i.e. larger galaxies live in more massive dark matter haloes. The $\eta$ is strongest for high mass luminous red galaxies (LRGs). Investigation of this relationship in hydrodynamical simulations suggests that, at a fixed $M_{*}$, satellite galaxies have a larger $\eta$ and greater scatter in the $M_{\mathrm{h}}$ and $r_{\mathrm{eff}}$ relationship compared to central galaxies., Comment: 23 pages, 16 figures

The utility of a new laser interferometric technique, inline coherent imaging, for real time keyhole depth measurement during laser welding is demonstrated on five important engineering alloys. The keyhole depth was measured at 200 kHz with a spatial resolution of 22 μm using a probe beam, which enters the keyhole coaxially with the process beam. Keyhole fluctuations limited average weld depth determination to a resolution on the order of 100 μm. Real time keyhole depth data are compared with the weld depths measured from the corresponding metallographic cross-sections. With the exception of an aluminium alloy, the technique accurately measured the average weld depth with differences of less than 5%. The keyhole depth growth rates at the start of welding are measured and compare well with order of magnitude calculations. The method described here is recommended for the real time measurement and control of keyhole depth in at least five different alloys.

Changing-look quasars are a newly-discovered class of luminous active galactic nuclei that undergo rapid ($\lesssim$10 year) transitions between Type 1 and Type 1.9/2, with an associated change in their continuum emission. We characterize the host galaxies of four faded changing-look quasars using broadband optical imaging. We use \textit{gri} images obtained with the Gemini Multi Object Spectrograph (GMOS) on Gemini North to characterize the surface brightness profiles of the quasar hosts and search for [O III] $\lambda4959,\lambda5007$ emission from spatially extended regions, or voorwerpjes, with the goal of using them to examine past luminosity history. Although we do not detect, voorwerpjes surrounding the four quasar host galaxies, we take advantage of the dim nuclear emission to characterize the colors and morphologies of the host galaxies. Three of the four galaxies show morphological evidence of merger activity or tidal features in their residuals. The three galaxies which are not highly distorted are fit with a single S\'ersic profile to characterize their overall surface brightness profiles. The single-S\'ersic fits give intermediate S\'ersic indices between the $n=1$ of disk galaxies and the $n=4$ of ellipticals. On a color-magnitude diagram, our changing-look quasar host galaxies reside in the blue cloud, with other AGN host galaxies and star-forming galaxies. On a color-S\'ersic index diagram the changing-look quasar hosts reside with other AGN hosts in the "green valley". Our analysis suggests that the hosts of changing-look quasars are predominantly disrupted or merging galaxies that resemble AGN hosts, rather than inactive galaxies., Comment: 20 pages, 5 figures

Drug-coated balloons (DCB) have been used to treat de novo small vessel coronary disease (SVD), with promising results and shorter dual antiplatelet therapy (DAPT) duration compared to drug-eluting stents (DES). We compared safety and effectiveness of the two treatments at 1 year.We reviewed 3,613 angioplasty cases retrospectively from 2011 to 2013 and identified 335 patients with SVD treated with device diameter of ≤2.5 mm. DCB-only angioplasty was performed in 172 patients, whereas 163 patients were treated with second-generation DES.DCB patients had smaller reference vessel diameter (2.22 ± 0.30 vs. 2.44 ± 0.19 mm, P 0.001) and received smaller devices (median diameter 2.25 vs. 2.50 mm, P 0.001) compared to the DES group. DES-treated vessels had larger acute lumen gain (1.71 ± 0.48 mm) than DCB (1.00 ± 0.53 mm, P 0.001). Half the patients had diabetes mellitus. While there were more patients presenting with acute coronary syndrome (ACS) in the DCB group (77.9% vs. 62.2%, P = 0.013), they received shorter DAPT (7.4 ± 4.7 vs. 11.8 ± 1.4 months, P 0.001) than the DES group. The 1-year composite major adverse cardiac event rate was 11.6% in the DCB arm and 11.7% in the DES arm (P = 1.000), with target lesion revascularization rate of 5.2% and 3.7%, respectively, (P = 0.601).In this high-risk cohort of patients, DCB-only angioplasty delivered good clinical outcome at 1 year. The results were comparable with DES-treated patients, but had the added benefit of a shorter DAPT regime.

Background and Objective During tissue ablation, laser light can be delivered with high precision in the transverse dimensions but final incision depth can be difficult to control. We monitor incision depth as it progresses, providing feedback to ensure that material removal occurs within a localized target volume, reducing the possibility of undesirable damage to tissues below the incision. Materials and Methods Ex vivo cortical and cancellous bone was ablated using pulsed lasers with center wavelengths of 1,064 and 1,070 nm, while being imaged in real-time using inline coherent imaging (ICI) at rates of up to 300 kHz and axial resolution of ∼6 µm. With real-time feedback, laser exposure was terminated before perforating into natural inclusions of the cancellous bone and verified by brightfield microscopy of the crater cross-sections accessed via side-polishing. Results ICI provides direct information about incision penetration even in the presence of intense backscatter from the pulsed laser and plasma emissions. In this study, ICI is able to anticipate structures 176 ± 8 µm below the ablation front with signal intensity 9 ± 2 dB above the noise floor. As a result, the operator is able to terminate exposure of the laser sparing a 50 µm thick layer of bone between the bottom of the incision to a natural inclusion in the cancellous bone. Versatility of the ICI system was demonstrated over a wide range of light–tissue interactions from thermal regime to direct solid-plasma transition. Conclusions ICI can be used as non-contact real-time feedback to monitor the depth of an incision created by laser ablation, especially in heterogeneous tissue where ablation rate is less predictable. Furthermore, ICI can image below the ablation front making it possible to stop laser exposure to limit unintentional damage to subsurface structures such as blood vessels or nervous tissue. Lasers Surg. Med. 44:249–256, 2012. © 2012 Wiley Periodicals, Inc.

Excessive glutamatergic neurotransmission, particularly when mediated by the N:-methyl-D-aspartate (NMDA) subtype of glutamate receptor, is thought to underlie neuronal death in a number of neurological disorders. Histamine has been reported to potentiate NMDA receptor-mediated events under a variety of conditions. In the present study we have utilized primary hippocampal neurone cultures to investigate the effect of mast cell-derived, as well as exogenously applied, histamine on neurotoxicity evoked by excessive synaptic activity. Exposure of mature cultures for 15 min to an Mg(2+)-free/glycine-containing buffer to trigger synaptic transmission through NMDA receptors, caused a 30-35% neuronal loss over 24 h. When co-cultured with hippocampal neurones, activated mast cells increased excitotoxic injury to 60%, an effect that was abolished in the presence of histaminase. Similarly, addition of histamine during magnesium deprivation produced a concentration-dependent potentiation (+ 60%; EC(50) : 5 microM) of neuronal death which was inhibited by sodium channel blockers and NMDA receptor antagonists, although this effect did not involve known histamine receptors. The histamine effect was further potentiated by acidification of the culture medium. Cultures 'preconditioned' by sublethal (5 min) Mg(2+) deprivation exhibited less neuronal death than controls when exposed to a more severe insult. NMDA receptor activation and the extracellular regulated kinase cascade were required for preconditioning neuroprotection. The finding that histamine potentiates NMDA receptor-mediated excitotoxicity may have important implications for our understanding of conditions where enhanced glutamatergic neurotransmission is observed in conjunction with tissue acidification, such as cerebral ischaemia and epilepsy.

Spreading depression (SD) has long been associated with the underlying pathophysiology of migraine. Evidence that the N-methyl-D-aspartate (NMDA) glutamate receptor (NMDA-R) is implicated in the generation and propagation of SD has itself been available for more than 15 years. However, to date, there are no reports of NMDA-R antagonists being developed for migraine therapy. In this study, an uncompetitive, pan-NMDA-R blocker, memantine, approved for clinical use, and two antagonists with selectivity for NMDA-R containing the NR2B subunit, (1S,2S)-1-(4-hydroxyphenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanol (CP-101,606) and (+/-)-(R*,S*)-alpha-(4-hydroxyphenyl)-beta-methyl-4-(phenylmethyl)-1-piperidine propanol (Ro 25-6981), were investigated to assess their protective effects against SD in the rat. Under isoflurane anesthesia, d.c. potential and the related cortical blood flow and partial pressure of O2 (pO2) were recorded simultaneously at separate cortical sites. Drugs (1, 3, and 10 mg/kg i.p.) were given 1 h or 30 min before KCl application to the brain surface. Core temperature and arterial pCO2,pO2, and pH measurements confirmed physiological stability. KCl induced 7.7+/-1.8 (mean+/-S.D.) SD events with d.c. amplitude of 14.9+/-2.8 mV. Memantine and CP-101,606 dose-dependently decreased SD event number (to 2.0+/-1.8 and 2.3+/-2.9, respectively) and SD amplitude at doses relevant for therapeutic use. Ro 25-6981 also decreased SD events significantly, but less effectively (to 4.5+/-1.6), without affecting amplitude. These results indicate that NR2B-containing NMDA receptors are key mediators of SD, and as such, memantine- and NR2B-selective antagonists may be useful new therapeutic agents for the treatment of migraine and other SD-related disorders (e.g., stroke and brain injury). Whether chronic, rather than acute, treatment may improve their efficacy remains to be determined.

Perturbation of normal survival mechanisms may play a role in a large number of disease processes. Glutamate neurotoxicity, particularly when mediated by the N-methyl-D-aspartate (NMDA) subtype of glutamate receptors, has been hypothesized to underlie several types of acute brain injury, including stroke. Several neurological insults linked to excessive release of glutamate and neuronal death result in tyrosine kinase activation, including p44/42 mitogen activated protein (MAP) kinase. To further explore a role for MAP kinase activation in excitotoxicity, we used a novel tissue culture model to induce neurotoxicity. Removal of the endogenous blockade by Mg2+ of the NMDA receptor in cultured hippocampal neurons triggers a self perpetuating cycle of excitotoxicity, which has relatively slow onset, and is critically dependent on NMDA receptors and activation of voltage gated Na+ channels. These injury conditions led to a rapid phosphorylation of p44/42 that was blocked by MAP kinase kinase (MEK) inhibitors. MEK inhibition was associated with protection against synaptically mediated excitotoxicity. Interestingly, hippocampal neurons preconditioned by a sublethal exposure to Mg(2+)-free conditions were rendered resistant to injury induced by a subsequently longer exposure to this insult; the preconditioning effect was MAP kinase dependent. The MAP kinase signaling pathway can also promote polypeptide growth factor mediated neuronal survival. MAP kinase regulated pathways may act to promote survival or death, depending upon the cellular context in which they are activated.

Original article can be found at: http://www.nature.com/bjp/index.html Copyright British Pharmacological Society and Nature Publishing Group. DOI: 10.1038/sj.bjp.0705945 [Full text of this article is not available in the UHRA]

Activation of the mitogen-activated protein kinase (MAPK/ERK) signal transduction pathway may mediate excitotoxic neuronal cell death in vitro and during ischemic brain injury in vivo. However, little is known, of the upstream regulation or downstream consequences of ERK activation under these conditions. Magnesium removal has been described to induce hyperexcitability and degeneration in cultured hippocampal neurones. Here, we show that neurotoxicity evoked by Mg2+ removal in primary hippocampal neurones stimulates ERK, but not p38, phosphorylation. Removal of Mg2+ also resulted in induction of the MAPK/ERK substrate mitogen- and stress-response kinase 1 (MSK1) and induced phosphorylation of the MSK1 substrate, the transcription factor cAMP response element binding protein (CREB). Neuronal death and phosphorylation of components in this cascade were inhibited by the Raf inhibitor SB-386023, by the MEK inhibitor U0126, or by the MSK1 inhibitors H89 and Ro318220. Importantly, this form of cell death was inhibited in hippocampal neurones cultured from MSK1-/- mice and inhibitors of Raf or MEK had no additive neuroprotective effect. Together, these data indicate that MSK1 is a physiological kinase for CREB and that this activity is an essential component of activity-dependent neuronal cell death.

Our understanding of the pathophysiological mechanisms of migraine remains poor despite the availability of clinically effective drugs and many years of research. Historically, two independent theories regarding the aetiology of headache were suggested: vascular and neuronal. However, recent data demonstrate that neuronal excitation modulates both the pial and meningeal circulation through a critical interaction with the trigeminal nerve, supporting the concept that the integration of neuronal and vascular information in the trigeminovascular network represents a key event in the aetiology of migraine.

In deep-penetration laser welding processes, the shape of the keyhole determines both the incoupling efficiency of the laser radiation and the dynamics of the melt flow. The instability in its shape is responsible for the generation of defects including pores and melt ejections and is therefore closely correlated to the weld quality. The shape of the keyhole and its dynamic behavior is strongly influenced by welding parameters and material properties.In order to detect the shape of the keyhole and its stability during the welding process, online diagnostics are of great interest. A low coherence interferometry method known as Inline Coherent Imaging (ICI) was shown to be a suitable means to measure the optical path length in the keyhole with a temporal resolution better than 1 ms [1]. Such systems take benefit of the fact, that the keyhole is filled with vapor and therefore transparent for a wide range of wavelengths. Especially the welding depth of stable processes can be measured with good accuracy. In unstable processes such as welding of aluminum-magnesium alloys, we observe strong fluctuations in the indicated depth. To investigate the correlation of measured fluctuations with keyhole instabilities, the laser-welding process was observed with a high speed X-ray transmission imaging system during the optical depth measurement. It was seen that an unstable keyhole generates significantly more pores then a stable keyhole. With the X-ray system, the keyhole depth and width were analyzed with a frame-rate of 1 kHz. The ICI system was used at a sampling rate of more than 10 kHz.The depth measurement results of both systems were correlated to the generation of pores, which are clearly visible in the X-ray records. Different signal analyzing methods for the ICI signal will be presented and discussed. We show that ICI allows detection of keyhole instabilities and with it pore formation and melt pool ejections.In deep-penetration laser welding processes, the shape of the keyhole determines both the incoupling efficiency of the laser radiation and the dynamics of the melt flow. The instability in its shape is responsible for the generation of defects including pores and melt ejections and is therefore closely correlated to the weld quality. The shape of the keyhole and its dynamic behavior is strongly influenced by welding parameters and material properties.In order to detect the shape of the keyhole and its stability during the welding process, online diagnostics are of great interest. A low coherence interferometry method known as Inline Coherent Imaging (ICI) was shown to be a suitable means to measure the optical path length in the keyhole with a temporal resolution better than 1 ms [1]. Such systems take benefit of the fact, that the keyhole is filled with vapor and therefore transparent for a wide range of wavelengths. Especially the welding depth of stable processes can be measured with good accuracy. In ...

Myelin-associated glycoprotein (MAG) is expressed on myelinating glia and inhibits neurite outgrowth from post-natal neurons. MAG has a sialic acid binding site in its N-terminal domain and binds to specific sialylated glycans and gangliosides present on the surface of neurons, but the significance of these interactions in the effect of MAG on neurite outgrowth is unclear. Here we present evidence to suggest that recognition of sialylated glycans is essential for inhibition of neurite outgrowth by MAG. Arginine 118 on MAG is known to make a key contact with sialic acid. We show that mutation of this residue reduces the potency of MAG inhibitory activity but that residual activity is also a result of carbohydrate recognition. We then go on to investigate gangliosides GT1b and GD1a as candidate MAG receptors. We show that MAG specifically binds both gangliosides and that both are expressed on the surface of MAG-responsive neurons. Furthermore, antibody cross-linking of cell surface GT1b, but not GD1a, mimics the effect of MAG, in that neurite outgrowth is inhibited through activation of Rho kinase. These data strongly suggest that interaction with GT1b on the neuronal cell surface is a potential mechanism for inhibition of neurite outgrowth by MAG.

Ion channels gated directly by cyclic nucleotides are required for the transduction of sensory signals in photoreceptor cells and olfactory cells. Cyclic nucleotide-gated (CNG) channels may also be expressed in the central nervous system because partial transcripts that share homology with CNG channels have been found therein. We have now isolated and cloned a full-length CNG channel cDNA from adult rat brain. The sequence is identical to that of the alpha-subunit originally found in the olfactory epithelium (CNCalpha3). In situ hybridization, using probes specific for the CNCalpha3 mRNA, suggest that this channel is expressed widely in the rat brain, albeit mostly at relatively low levels. Certain neuronal populations, however, such as deep cerebellar nuclei, olfactory bulb mitral cells and cerebellar Purkinje neurons, appeared specially enriched. The study demonstrates for the first time that a full-length CNG channel mRNA is expressed in the brain, supporting the possibility that CNG channels are involved in central neural communication and plasticity. The sequence reported in this paper has been deposited in the GenBank data base (accession no. AF126808).

The high-energy collision-induced dissociation (CID) spectra of the [M+H]+ and [M-H]- ions of substance P fragment 6–11, the retropeptide and the corresponding peptoid and retropeptoid were compared. The CID spectra of the [M+H]+ ion of the corresponding (retro)peptide and (retro)peptoid exhibit both B- and Y″-type sequence ions at identicalm/z-values. The differences in the relative abundances of these sequence ions, however, can be related to structural characteristics of the compounds. The fragmentation behaviour of theN-substituted immonium ions differs from that of the immonium ions derived from common amino acids by showing a preferential loss of a CH2=NH imine molecule. This dominant fragmentation reaction is suppressed in the CID spectrum of theN-substituted glutamine immonium ion in favour of a less energy demanding cyclization reaction involving the loss of NH3. The CID spectra of the [M-H]- ions of both peptides and peptoids show C-type ions, which appeared to be more abundant in the spectra of the peptides than in those of the peptoids.C-Terminal ″Z- and Y-type ions are characteristic of the peptides, whereas the peptoid spectra show relatively abundantN-terminal ″B-type ions. Both [M+H]+ and [M-H]- ion CID spectra show loss of amino acid-specific side-chains, which occurs as radical loss in the case of peptides and as molecular loss in peptoids. © 1997 by John Wiley & Sons, Ltd.

We achieve in situ, micron-scale tracking of laser machining through a galvo-telecentric beam delivery system using coherent imaging. We collect both high speed intrapulse and interpulse morphology changes as well as overall sweep-to-sweep depth penetration.

Controlling depth aspects during practical laser materials processing has remained a challenge for many years. Inline Coherent Imaging is an emerging technique that may hold important answers to "deep" questions for industry and basic science.

In applications ranging from noncontact microsurgery to semiconductor blind hole drilling, precise depth control of laser processing is a major challenge. Even expensive a priori characterization cannot compensate for material heterogeneity and stochasticity inherent to the ablation process. Here we use in situ depth imaging to guide the machining process in real time. W e image along the machining beam axis at high speeds (up to 300 kHz) to provide real-time feedback, even in high aspect ratio holes. The in situ metrology is based on coherent imaging (similar to optical coherence tomography) and is practical for a wide-range of light sources and machining processes (e.g., thermal cutting or ultrafast nonlinear ablation). Coherent imaging has a high dynamic range (> 60 dB) and strongly rejects incoherent signals allowing weak features to be observed in the presence of intense machining light and plasmas. High axial resolution (∼10 μm) requires broadband imaging light but the center wavelength can be chosen appropriate to the application. Infrared light (wavelength: 1320 ± 35 nm) allows simultaneous monitoring of both surface and subsurface interfaces in non-absorbing materials like tissue and semiconductors. By contrast, silicon based detector technology can be used with near infrared imaging light (805 ± 25 nm) enabling high speed acquisition and low cost implementation.

We observe sample morphology changes in real time (24 kHz) during and between percussion drilling pulses by integrating a low-coherence microscope into a laser micromachining platform. Nonuniform cut speed and sidewall evolution in stainless steel are observed to strongly depend on assist gas. Interpulse morphology relaxation such as hole refill is directly imaged, showing dramatic differences in the material removal process dependent on pulse duration/peak power (micros/0.1 kW, ps/20 MW) and material (steel, lead zirconate titanate PZT). Blind hole depth precision is improved by over 1 order of magnitude using in situ feedback from the imaging system.

The stochastic effects of assist gas in QCW and pulsed laser machining (percussion drilling) in steel are measured with a novel in situ high speed low coherence imaging system. Real-time imaging is delivered coaxially with machining energy and assist gas revealing relaxation and melt flow dynamics over microsecond timescales and millimeter length scales with ~10 micrometer resolution. Direct measurement of cut rate and repeatability avoids post cut analysis and iterative process development. Feedback from the imaging system can be used to overcome variations in relaxation and guides blind hole cutting.

It has been shown that 30 ns FWHM duration pulses from a MOPA fiber laser (wavelength: 1064 nm) cleanly micromachines silicon with little cracking or heat-affected zone 1 . In this paper, we show that similar results can be achieved using a 1070 nm quasi-continuous wave laser pulsed with a 6.6 μs duration (average power: 2.8 W) in combination with coaxially delivered nitrogen assist gas. The holes are cut at a 5 kHz repetition rate with a resulting diameter on the order of 15 μm and an etch rate of up to 18 μm/pulse. Hole size is increased for longer pulses and the heat-affected zone broadens to greater than 25 μm with no assist gas. By combining low coherence microscopy with machining, we depth image the machining front and obtain in situ images during and after the drilling process showing rich cut dynamics in real time.

A key issue in laser surgery is the inability for the human operator to stop the laser irradiation in time while cutting/ablating delicate tissue layers. In the present work, we forward-image through the laser machining front in complex biological tissue (dense bovine bone) to monitor the incision's approach to subsurface interfaces in real-time (47-312 kHz line rate). Feedback from imaging is used to stop the drilling process within 150 micron of a targeted interface. This is accomplished by combining the high temporal and spatial resolution of infrared optical coherence tomography (OCT) with a robust, turn-key, high brightness fiber laser. The high sensitivity of the imaging system (~100 dB) permit imaging through the rapidly changing beam path even with the additional scattering caused by the thermal cutting process. In spectral-domain OCT, the imaging acquisition period is easily locked to the machining laser exposure. Though motion-induced artifacts reduce interface contrast, they do not introduce incorrect depth measurements as found in other OCT variants. Standard tomography imaging of the tissue (B-scans) is also recorded in situ before and after laser processing to highlight morphology changes.

Optical coherence imaging can measure hole depth in real-time (> 20 kHz) during laser drilling without being blinded by intense machining light or incoherent plasma emissions. Rapid measurement of etch rate and stochastic melt relaxation makes these images useful for process development and quality control in a variety of materials including metals, semiconductors and dielectrics. The ability to image through the ablation crater in materials transparent to imaging light allows the guidance of blind hole cutting even with limited a priori knowledge of the sample.Significant improvement in hole depth accuracy with the application of manual feedback from this imaging has been previously demonstrated [1]. However, the large quantity of raw data and computing overhead are obstacles for the application of coherent imaging as a truly automatic feedback mechanism. Additionally, the high performance components of coherent imaging systems designed for its traditional application in biological imaging are costly and may be unnecessary for materials processing. In this work, we present a coherent imaging system design that costs less than a fifth of comparable commercial products. We also demonstrate streamlined image processing suited for automated feedback that increases processing speed by two orders of magnitude.Optical coherence imaging can measure hole depth in real-time (> 20 kHz) during laser drilling without being blinded by intense machining light or incoherent plasma emissions. Rapid measurement of etch rate and stochastic melt relaxation makes these images useful for process development and quality control in a variety of materials including metals, semiconductors and dielectrics. The ability to image through the ablation crater in materials transparent to imaging light allows the guidance of blind hole cutting even with limited a priori knowledge of the sample.Significant improvement in hole depth accuracy with the application of manual feedback from this imaging has been previously demonstrated [1]. However, the large quantity of raw data and computing overhead are obstacles for the application of coherent imaging as a truly automatic feedback mechanism. Additionally, the high performance components of coherent imaging systems designed for its traditional application in biological imaging are costly and...

In the present studies, the mechanisms underlying the inhibitory actions of lipocortin-1 on the pyrogenic and thermogenic properties of cytokines were investigated. Central (icv) injection of corticotropin-releasing factor (CRF, 4.7 micrograms) or the recombinant cytokines interleukin (IL)-1 alpha (50 ng), IL-1 beta (5 ng), IL-6 (20 ng), IL-8 (20 ng), or tumor necrosis factor-alpha (TNF-alpha, 1 microgram) in conscious rats produced significant increases in resting oxygen consumption (VO2, 13-26%) and colonic temperature (0.7-1.6 degrees C) within 2 h postinjection. Administration (icv) of a recombinant fragment (NH2-terminus, 1-188 amino acids) of human lipocortin-1 (1.2 micrograms) produced small increases in VO2 (< 5%) and body temperature (< 0.3 degrees C). Pretreatment (-5 min) with lipocortin-1 significantly attenuated the thermogenic and pyrogenic effects of centrally injected IL-1 beta (80% inhibition), IL-6 (60%), IL-8 (80%), or CRF (60%). However, pretreatment with lipocortin-1 failed to modify the actions of IL-1 alpha or TNF-alpha. We have previously demonstrated that the pyrogenic and thermogenic effects of IL-1 beta, IL-6, and IL-8 are dependent on the central actions of CRF, whereas IL-1 alpha and TNF-alpha act independently of CRF. Fever and thermogenesis induced by all of these cytokines (with the exception of IL-8) can also be prevented by administration of a cyclooxygenase inhibitor. The data presented here suggest that the potent antipyretic effects of lipocortin-1 may result from inhibition of the release or actions of CRF rather than modulation of eicosanoid synthesis.

By integrating coherent imaging (optical coherence tomography) into an ultrafast machining platform, we directly monitor surface and subsurface changes in sample morphology due to the laser ablation and subsequent relaxation between laser pulses.

We demonstrate in situ observation of percussion drilling in stainless steel at axial rates of 40 kHz. The melt cycle is directly observed and imaging feedback is used to improve cut accuracy.

The presence and localization of endogenous lipocortin-1 (LC-1, a protein which has been proposed to mediate the anti-inflammatory actions of the glucocorticoids) was studied by immunohistochemical techniques in rat brain and pituitary. A polyclonal antiserum specific for a fragment of lipocortin-1 (alpha alpha 1-188) was used to visualize immunoreactive LC-1 (iLC-1) in both neuronal and non-neuronal cell structures. Neuronal staining, which was independent of microtubular axonal transport mechanisms (in that it was not affected by blockade of axonal transport), was found in varicose nerve fibres in various regions of the brain. In addition, iLC-1 was found in the cytoplasm of neuronal cells throughout the brain. Of all brain regions which showed iLC-1, only the hippocampal neurons showed a reduced staining intensity after adrenalectomy. However, iLC-1 was not affected by dexamethasone treatment. Non-neuronal iLC-1 was found in ependymocytes lining the cerebral ventricles and aqueduct. In addition, iLC-1 was found in tanycytes in all circumventricular organs studied and in the ventral walls of the third ventricle, where some of the branching tail processes appeared to envelop local capillaries and neuronal cell bodies. A tancycyte-mediated release of LC-1 from varicose nerve fibres into the portal vasculature is proposed.

We present a case of a chronic total occlusion that required the use of a 5 Fr in a 6 Fr guiding catheter. The 5 Fr in a 6 Fr guiding catheter method allows ultra-deep seating to increase backup support and assist stent delivery. This technique was paramount in this case, as there were technical challenges with failure to track drug-eluting stents due to vessel tortuosity and lack of support.

We demonstrate simultaneous drilling and coaxial depth imaging of holes in stainless steel at axial rates of 46 kHz. Depth measurement with 6 mum resolution is performed using the machining beam via Fourier-domain interferometry.

Lipocortins form a group of proteins that have been proposed as mediators of the anti-inflammatory actions of glucocorticoids. Intracerebroventricular injection of a recombinant fragment of lipocortin 1 (NH2-terminal 1-188) caused dose-dependent (0.4-1.2 micrograms) reductions in the acute increases in colonic temperature and oxygen consumption, which occurred in response to central injections of recombinant interleukin 1 beta and gamma-interferon in conscious rats. In contrast the lipocortin fragment did not affect the response to prostaglandin E2, and its activity was prevented by heat treatment or by pretreatment of animals with polyclonal antiserum raised to the fragment. Central injection of antiserum significantly enhanced the thermogenic responses to interleukin 1 beta in rats treated with dexamethasone without affecting the responses in normal animals. These results support a physiological role for lipocortin in the central effects of glucocorticoids.

Scanning optics enable many laser applications in manufacturing because their low inertia allows rapid movement of the process beam across the sample. We describe our method of inline coherent imaging for real-time (up to 230 kHz) micron-scale (7–8 µm axial resolution) tracking and control of laser machining depth through a scanning galvo-telecentric beam delivery system. For 1 cm trench etching in stainless steel, we collect high speed intrapulse and interpulse morphology which is useful for further understanding underlying mechanisms or comparison with numerical models. We also collect overall sweep-to-sweep depth penetration which can be used for feedback depth control. For trench etching in silicon, we show the relationship of etch rate with average power and scan speed by computer processing of depth information without destructive sample post-processing. We also achieve three-dimensional infrared continuous wave (modulated) laser machining of a 3.96 × 3.96 × 0.5 mm3 (length × width × maximum depth) pattern on steel with depth feedback. To the best of our knowledge, this is the first successful demonstration of direct real-time depth monitoring and control of laser machining with scanning optics.

The human hyperpolarization-activated cyclic nucleotide-gated 1 (hHCN1) subunit was heterologously expressed in mammalian cell lines (CV-1 and CHO) and its properties investigated using whole-cell patch-clamp recordings. Activation of this recombinant channel, by membrane hyperpolarization, generated a slowly activating, noninactivating inward current. The pharmacological properties of hHCN1-mediated currents resembled those of native hyperpolarization-activated currents (I(h)), that is, blockade by Cs(+) (99% at 5 mm), ZD 7288 (98% at 100 microm) and zatebradine (92% at 10 microm). Inhibition of the hHCN1-mediated current by ZD 7288 was apparently independent of prior channel activation (i.e. non-use-dependent), whereas that induced by zatebradine was use-dependent. The VR1 receptor antagonist capsazepine inhibited hHCN1-mediated currents in a concentration-dependent (IC(50)=8 microm), reversible and apparently non-use-dependent manner. This inhibitory effect of capsazepine was voltage-independent and associated with a leftward shift in the hHCN1 activation curve as well as a dramatic slowing of the kinetics of current activation. Elevation of intracellular cAMP or extracellular K(+) significantly enhanced aspects of hHCN1 currents. However, these manipulations did not significantly affect the capsazepine-induced inhibition of hHCN1. The development of structural analogues of capsazepine may yield compounds that could selectively inhibit HCN channels and prove useful for the treatment of neurological disorders where a role for HCN channels has been described.

Calcitonin gene-related peptide (CGRP) released from trigeminal primary afferents has been implicated in the pathophysiology of migraine. Here, we have used an in vitro slice preparation to investigate its release from nerve terminals in the rat trigeminal nucleus caudalis. Extracellular-calcium dependent CGRP release was stimulated by both capsaicin and neuronal depolarization with KCl. The capsaicin (1M)-evoked CGRP release was blocked by capsazepine and was also attenuated in the presence of the cyclooxygenase inhibitor, indomethacin, an effect that was reversed when slices were stimulated with capsaicin in the presence of the cyclooxygenase metabolite, prostaglandin E2. Taken together, these data further highlight the importance of prostaglandins as enhancers of neuropeptide release and suggest that CGRP released from the central terminals of trigeminal neurones has the potential to be involved in the transmission of nociceptive information of relevance to migraine headache. © 2004 Elsevier Ireland Ltd. All rights reserved.

Corticotropin-releasing factor (CRF) receptors are members of the superfamily of G-protein coupled receptors that utilise adenylate cyclase and subsequent production of cAMP for signal transduction in many tissues. Activation of cAMP-dependent pathways, through elevation of intracellular cAMP levels is known to promote survival of a large variety of central and peripheral neuronal populations. Utilising cultured primary rat central nervous system neurons, we show that stimulation of endogenous cAMP signalling pathways by forskolin confers neuroprotection, whilst inhibition of this pathway triggers neuronal death. CRF and the related CRF family peptides urotensin I, urocortin, and sauvagine, which also induced cAMP production, prevented the apoptotic death of cerebellar granule neurons triggered by inhibition of phosphatidylinositol kinase-3 pathway activity with LY294002. These effects were negated by the highly selective CRF-R1 antagonist CP154,526. CRF even conferred neuroprotection when its application was delayed by up to 8 h following LY294002 addition. The CRF peptides also protected cortical and hippocampal neurons against death induced by β-amyloid peptide (1-42), in a CRF-R1 dependent manner. In separate experiments, LY294002 reduced neuronal protein kinase B activity while increasing glycogen synthase kinase-3, whilst CRF (and related peptides) promoted phosphorylation of glycogen synthase kinase-3 without protein kinase B activation. Taken together, these results suggest that the neuroprotective activity of CRF may involve cAMP-dependent phosphorylation of glycogen synthase kinase-3.

Activation of the mitogen-activated protein kinase (MAPK/ERK) signal transduction pathway may mediate excitotoxic neuronal cell death in vitro and during ischemic brain injury in vivo. However, little is known, of the upstream regulation or downstream consequences of ERK activation under these conditions. Magnesium removal has been described to induce hyperexcitability and degeneration in cultured hippocampal neurones. Here, we show that neurotoxicity evoked by Mg2+ removal in primary hippocampal neurones stimulates ERK, but not p38, phosphorylation. Removal of Mg2+ also resulted in induction of the MAPK/ERK substrate mitogen- and stress-response kinase 1 (MSK1) and induced phosphorylation of the MSK1 substrate, the transcription factor cAMP response element binding protein (CREB). Neuronal death and phosphorylation of components in this cascade were inhibited by the Raf inhibitor SB-386023, by the MEK inhibitor U0126, or by the MSK1 inhibitors H89 and Ro318220. Importantly, this form of cell death was inhibited in hippocampal neurones cultured from MSK1-/- mice and inhibitors of Raf or MEK had no additive neuroprotective effect. Together, these data indicate that MSK1 is a physiological kinase for CREB and that this activity is an essential component of activity-dependent neuronal cell death.

Pfizer has developed and launched eletriptan, a 5-HT1B/1D agonist, for the potential treatment of migraine with and without aura. Eletriptan has 6-fold greater affinity for the 5-HT1D receptor than sumatriptan, and a 3-fold greater affinity for the 5-HT1B receptor [249570]. Eletriptan pharmacology has also been evaluated in vitro in comparison with zolmitriptan (AstraZeneca plc) and naratriptan (GlaxoSmithKline plc) [290116].

Inhibition of the p38 mitogen-activated protein kinase (MAP Kinase) pathway reduces acute ischemic injury in vivo, suggesting a direct role for this signaling pathway in a number of neurodegenerative processes. The present study was designed to evaluate further the role of p38 MAP Kinase in acute excitotoxic neuronal injury using the selective p38 inhibitor SB-239063 (trans-1-(4hydroxycyclohexyl)-4-(fluorophenyl)-5-(2-methoxy-pyrimidin-4-yl) imidazole). Unlike the widely used p38 inhibitor, SB-203580 (4-(4-Fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole), this second generation p38 inhibitor more selectively inhibits p38 MAP Kinase without affecting the activity of other MAP Kinase signaling pathways and provides a more accurate means to selectively assess the role of p38 in excitotoxicity that has not been previously possible. SB-239063 provided substantial protection against cell death induced by either oxygen glucose deprivation (OGD) or magnesium deprivation in cultured neurons. The ability of this compound to block excitotoxicity was not due to direct inhibition of N-methyl-D-aspartate (NMDA) receptor-mediated currents as SB-239063 did not alter NMDA electrophysiological responses. SB-239063 did not protect against a severe excitotoxic insult induced by 60-min exposure to NMDA. However, when tested against a less severe, brief (5 min) NMDA exposure, p38 inhibition provided substantial protection. These data demonstrate that inhibition of p38 MAP Kinase can confer neuroprotection in vitro against mild but not severe excitotoxic exposure, and suggests that other additional pathways/mechanism(s) may be involved in severe excitotoxic cell death.

Although new affordable high-power laser technologies enable many processing applications in science and industry, depth control remains a serious technical challenge. In this Letter we show that inline coherent imaging (ICI), with line rates up to 312 kHz and microsecond-duration capture times, is capable of directly measuring laser penetration depth, in a process as violent as kW-class keyhole welding. We exploit ICI's high speed, high dynamic range, and robustness to interference from other optical sources to achieve automatic, adaptive control of laser welding, as well as ablation, achieving 3D micron-scale sculpting in vastly different heterogeneous biological materials.

Salmonella typhimurium causes an invasive disease in mice that has similarities to human typhoid, with key roles for cytokines and possibly also inducible nitric oxide synthase (iNOS), in mediating host responses to infection. In this paper we demonstrate that iNOS mRNA, protein and enzyme activity is induced within spleens and livers of infected mice as early as 5 h post-infection. Immunohistochemistry and in situ hybridization indicated that iNOS expression occurs predominantly in macrophages in localized, discrete foci in the infected organs. iNOS activity in spleen and liver was not detectable in uninfected control mice. The presence of mRNA encoding pro-inflammatory cytokines (TNFalpha, IL-1beta and IFNgamma) in infected organs was measured using RT-PCR, all three being present from 2 h post-infection onwards, but not before. These data show that there is a very early host response to S. typhimurium infection in mice, limited to foci within the infected organs.

Fractalkine is a recently identified chemokine that exhibits cell adhesion and chemoattractive properties. It represents a unique member of the chemokine superfamily because it is located predominantly in the brain in which it is expressed constitutively on specific subsets of neurons. To elucidate the possible role of neuronally expressed fractalkine in the inflammatory response to neuronal injury, we have analyzed the regulation of fractalkine mRNA expression and protein cleavage under conditions of neurotoxicity. We observed that mRNA encoding fractalkine is unaffected by experimental ischemic stroke (permanent middle cerebral artery occlusion) in the rat. Similarly, in vitro, levels of fractalkine mRNA were unaffected by ensuing excitotoxicity. However, when analyzed at the protein level, we found that fractalkine is rapidly cleaved from cultured neurons in response to an excitotoxic stimulus. More specifically, fractalkine cleavage preceded actual neuronal death by 2-3 hr, and, when evaluated functionally, fractalkine represented the principal chemokine released from the neurons into the culture medium upon an excitotoxic stimulus to promote chemotaxis of primary microglial and monocytic cells. We further demonstrate that cleavage of neuron-derived, chemoattractive fractalkine can be prevented by inhibition of matrix metalloproteases. These data strongly suggest that dynamic proteolytic cleavage of fractalkine from neuronal membranes in response to a neurotoxic insult, and subsequent chemoattraction of reactive immune cells, may represent an early event in the inflammatory response to neuronal injury.

The observation that the free radical nitric oxide (NO) acts as a cell signaling molecule in key physiological processes such as regulation of blood pressure and immunological host-defense responses is probably one of the most important and exciting findings made in biology in the last decade. Likewise, in the brain NO has been implicated in a number of fundamental processes, including memory formation, sexual behavior and the control of cerebral blood flow. This has radically altered the accepted dogma of brain physiology and has placed NO at the center stage of neuroscience research. Evidence suggests that some of the actions of NO in the brain may be intimately linked to those of the classic excitatory neurotransmitter glutamate. The historical view that aberrations in glutamate signal transduction may underlie central neurodegeneration following, for example, cerebral ischemia, has implicated NO, by default, as a potential mediator of neuronal death. Indeed, with the advent of potent and specific compounds that interact with NO synthesizing (NOS) enzymes and with the NO signaling cascade, there is now ample evidence to suggest that NO can mediate neurodegeneration, although its involvement is paradoxical. Its cerebrovascular effects may act to limit ischemic damage by preserving tissue perfusion and preventing platelet aggregation, while NO produced in the parenchyma, either directly following the ischemic insult or at a later stage as part of a neuroinflammatory response, may be deleterious to the outcome of ischemia. Nonetheless, significant efforts are made into the potential therapeutic use of chemical NO donors and specific NOS inhibitors in the treatment of cerebral ischemia and other central neurodegenerative disorders. Here, the latest concepts and developments in our understanding of the role of NO in cerebral ischemic neurodegeneration are discussed.

We achieve high aspect-ratio laser ablation of silicon with a strong nonlinear dependence on pulse duration while using a power density 10(6) times less than the threshold for typical multiphoton-mediated ablation. This is especially counter-intuitive as silicon is nominally transparent to the modulated continuous wave Yb:fiber laser used in the experiments. We perform time-domain finite-element simulations of thermal dynamics to investigate thermo-optical coupling and link the observed machining to an intensity-thresholded runaway thermo-optically nonlinear process. This effect, cascaded absorption, is qualitatively different from ablation observed using nanosecond-duration pulses and is general enough to potentially facilitate high-quality, high aspect-ratio, and economical processing of many materials.

The ex vivo tissue concentration of nitrite and nitrate (NOx) was found to correlate closely with the activity of nitric oxide synthase (NOS; EC 1.14.13.39) in various brain regions. Systemic administration of the nonselective NOS inhibitor N omega-nitro-L-arginine (L-NA) at doses that completely inhibited both central and peripheral NOS, depleted whole-brain and CSF NOx by up to 75% but had no effect on plasma NOx. Selective inhibition of central NOS by intracerebroventricular administration of L-NA methyl ester produced similar decreases in levels of whole-brain NOx. A residual concentration of NOx of 10-15 microM remained in all brain regions even after complete inhibition of brain NOS. Brain NOx content decreased rapidly and in parallel with the inhibition of brain NOS. The ex vivo measurement of levels of brain NOx was found to reflect the in vivo efficacy of several different types of NOS inhibitor: L-NA, N omega-monomethyl-l-arginine, and 7-nitroindazole. Intraperitoneal administration of the NOS substrate L-arginine increased brain NOx concentrations by up to 150% of control values. These results demonstrate that the ex vivo measurement of levels of brain tissue NOx is a rapid, reliable, and straightforward technique to determine NOS activity in vivo. This method can be used to assess both the regional distribution and the degree of inhibition of NOS activity in vivo.

Infection by lentiviruses such as human immunodeficiency virus (HIV) and Maedi-Visna virus (MVV) is associated with neurodegenerative disorders. We have investigated the neurotoxic mechanisms of a synthetic peptide of transactivating protein tat of MVV in striatal neuronal cultures. Tat peptide (but not control peptide) caused neuronal death, without affecting glial viability, in a time- and dose-dependent manner. Significant neuronal death was not observed until 6-8 h after tat peptide application (2.35-2350 nM), whereas half maximal and maximal cell death was observed after 12 and 24 h respectively. Tat peptide neurotoxicity could be partially inhibited by blockade of either N-methyl-D-aspartate (NMDA)- or non-NMDA receptors, suggesting that excessive neuroexcitation by glutamate or its analogues may contribute to tat-neurotoxicity. Furthermore, when both these glutamate receptor subtypes were blocked simultaneously, an increased degree of neuroprotection was observed. Finally, tat peptide toxicity was also reduced by blockade of L-type calcium channels. Calcium imaging revealed that intracellular calcium increases slowly upon tat application, predominantly due to entry of extracellular calcium. These results indicate that cellular calcium entry through voltage-gated calcium channels following activation of both NMDA and non-NMDA receptors, and subsequent accumulation of intracellular calcium may contribute to the neuronal death induced by tat protein.

This study investigated the involvement of corticotrophin-releasing factor (CRF) in acute neuronal damage induced by focal cerebral ischaemia or pharmacological activation of NMDA receptors in the rat brain. Intracerebroventricular injection of a CRF receptor antagonist (alpha-helical CRF9-41), markedly inhibited ischaemic (61%) and excitotoxic (41%) brain damage. Peripheral injection of a glucocorticoid antagonist (RU38486) did not affect ischaemic damage. Ischaemic and excitotoxic damage caused increased hypothalamic concentrations of CRF. These data indicate that CRF mediates ischaemic and excitotoxic neuronal damage in the rat, but that this effect is not dependent on glucocorticoids.