It is well documented that reactive oxygen species (ROS) affects neurodegeneration in the brain. Several studies also implicate ROS in the regulation of synapse function and learning and memory processes, although the precise source of ROS generation within these contexts remains to be further explored. Here we show that postsynaptic superoxide generation through PKCζ-activated NADPH oxidase 2 (NOX2) is critical for long-term depression (LTD) of synaptic transmission in the CA1–Shaffer collateral synapse of the rat hippocampus. Specifically, PKCζ-dependent phosphorylation of p47phox at serine 316, a NOX2 regulatory subunit, is required for LTD but is not necessary for long-term potentiation (LTP). Our data suggest that postsynaptic p47phox phosphorylation at serine 316 is a key upstream determinant for LTD and synapse weakening.
Synapse weakening is part of a group of physiological processes referred to as synaptic plasticity, which govern changes in synaptic function in response to neuronal activity, and are thought to represent the cellular and molecular mechanisms of learning and memory1. On the other hand, aberrant activation of synapse weakening signalling pathways has been reported in several Alzheimer's disease (AD) models2,3,4, suggesting that these signalling pathways represent a crucial interplay between physiology and the onset of disease-associated pathophysiology. Mounting evidence suggests that apoptotic signalling cascades, including caspase-3 and glycogen synthase kinase 3β (GSK-3β) activation, are centrally involved in physiological and pathophysiological forms of synapse weakening, manifest through postsynaptic α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) endocytosis and long-term depression (LTD) of synaptic transmission4,5,6. However, how these signals are first initiated is unknown.
Reactive oxygen species (ROS) are not only well-known upstream regulators of neuronal apoptosis7 and neurodegenerative signals8 but are also known to play important regulatory roles in aspects of neuronal physiology, including synaptic plasticity and synapse weakening9,10,11,12,13. ROS can originate from numerous sources to affect synaptic plasticity, including presynaptic neurons, postsynaptic neurons and microglia11, 13. However, to date there has been no characterization or elucidation of the precise mechanisms of ROS generation that regulate synaptic plasticity.
Here, we examined the production of postsynaptic ROS during LTD, revealing a key role for postsynaptic ROS production via NADPH oxidase 2 (NOX2). Crucially, we find that the activity of postsynaptic protein kinase C zeta (PKCζ) is also required for LTD and identify the phosphorylation of p47phox at serine 316 as a necessary step in this pathway. Therefore, our results uncover a role for a specific postsynaptic ROS production pathway in activity-dependent synapse weakening.
Superoxide ions are one of the primary forms of ROS and are known to be elevated in neurons following the activation of N-methyl-d-aspartate receptors (NMDARs)14,15,16. We therefore hypothesized that intra-neuronal superoxide radicals are upstream regulators of NMDAR-dependent forms of synaptic plasticity. To address this, we analysed the effects of superoxide dismutase (SOD), a class of endogenous enzymes that catalyse superoxide dismutation17, on an NMDAR-dependent form of LTD in rat hippocampal acute slices18. Accordingly, whilst application of low-frequency electric stimulation (LFS) during whole-cell patch clamp recording readily induced LTD in CA1–Schaffer collateral synapses (52.8 ± 9.0% of baseline, p = 0.002 vs. control input, Fig. 1a), postsynaptic infusion of SOD (300 units/ml) through the patch pipette blocked LTD (89.5 ± 8.7% of baseline, p = 0.115 vs. control input, Fig. 1b). To determine whether the superoxide radicals involved in this form of LTD could originate from an extracellular source, such as microglia13, we bath applied SOD. Given that SOD has poor membrane permeability19, extracellular bath application of the enzyme will catalyse extracellular superoxide dismutation without affecting intracellularly generated superoxide. Extracellular SOD application had no effect on LTD (57.5 ± 9.9% of baseline, p = 0.004 vs. control input, Fig. 1c), suggesting that postsynaptic intracellular superoxide, specifically, is critical for LTD expression. Since hydrogen peroxide (H2O2), another ROS, can be a product of SOD catalysis of superoxide dismutation and is also implicated in synaptic plasticity20, we tested whether H2O2 is required for LTD. Postsynaptic infusion of catalase (300 units/ml), an enzyme that catalyses the decomposition of H2O2 to water and oxygen, had no effect on LTD (66.8 ± 4.7% of baseline, p = 0.002 vs. control input, Fig. 1d). We also tested whether H2O2 may be inhibiting NMDAR-LTD, which could provide an alternative explanation for the inhibition of LTD induced by SOD injection. To test this, we postsynaptically injected SOD along with catalase, thereby scavenging the H2O2 product of the SOD reaction. In this experiment, LTD was also blocked (SOD+catalase, 90.1 ± 4.8% of baseline, p = 0.150 vs. baseline, Fig. 1e). In bath application experiments, SOD and catalase co-treatment did not affect LTD (SOD+catalase, 68.3 ± 1.2% of baseline, p < 0.001 vs. baseline, Fig. 1f). Together, these results suggest that postsynaptic intracellular superoxide is required for NMDAR-dependent hippocampal LTD and that H2O2 itself is neither required for nor an inhibitor of LTD.
Fig. 1: Postsynaptic superoxide is required for LTD expression.
While several ROS-inducing mechanisms are present in neurons, emerging evidence suggests that superoxide production post-NMDAR activation is catalysed by NADPH oxidase (NOX)15, 16. NOX is a membrane-bound enzymatic complex responsible for the production of ROS and is present in neurons where it can localize to synapses21. Specifically, NOX2, commonly known as the prototypical NOX, has been suggested as the primary regulator of NMDAR-activated superoxide generation15. While NOX1–4 are reported to be expressed in the brain, NOX3 constitutively generates superoxide without stimulation22 and NOX4 constitutively produces H2O223, an ROS that appears not to be involved in LTD. We therefore predicted that postsynaptic NOX1 and 2 are the most likely candidates for superoxide production during LTD. To test a requirement for these NOX isoforms for LTD, we utilized postsynaptic infusion of NOX inhibitors. Postsynaptic infusion of AEBSF (20 µM), a non-selective inhibitor of NOX24, blocked LTD expression in CA1 neurons of acute hippocampal slices (90.4 ± 6.0% of baseline, p = 0.146 vs. control input, Fig. 2a), whilst postsynaptic infusion of the specific NOX1 inhibitor ML-171 (3 μM)25 had no effect on LTD (58.9 ± 8.6% of baseline, p = 0.006 vs. control input, Fig. 2b). Postsynaptic infusion of apocynin (100 µM), a putative but non-selective inhibitor of NOX226, significantly impaired LTD expression (88.7 ± 8.3% of baseline, p = 0.583 vs. control input, Fig. 2c). In contrast, bath perfusion of apocynin after LTD induction failed to reverse the expression of LTD, indicating that NOX2 is not required for LTD maintenance (apocynin perfusion: 68.8 ± 4.0% of baseline vs. control: 78.9 ± 3.4% of baseline, p = 0.078, Supplementary Fig. S1). Together, these data are suggestive of a specific role for the NOX2 isoform of NOX in the regulation of LTD induction.
Fig. 2: Neuronal NADPH oxidase 2 (NOX2) is required for LTD expression.
Owing to the limited pharmacological selectivity and off-target effects of available NOX inhibitors, we next utilized short hairpin RNA (shRNA) to knockdown either NOX1 or 2 expression (Fig. 2d). Biolistic transfection of NOX1 or NOX2 shRNA into CA1 neurons of organotypic cultured hippocampal slices was performed. LTD was readily inducible in cells transfected with NOX1 shRNA (NOX1 shRNA: 54.9 ± 4.7% of baseline; untransfected cells: 61.7 ± 3.5% of baseline, p = 0.280, Fig. 2e). In contrast, LTD was abolished in neurons transfected with NOX2 shRNA (NOX2 shRNA: 87.3 ± 11.5% of baseline; untransfected cells: 50.7 ± 7.4% of baseline, p = 0.028, Fig. 2f). Neither NOX1 nor NOX2 shRNA influenced the basal state of glutamatergic synaptic transmission, as measured by AMPAR-mediated excitatory postsynaptic current (EPSCAMPA; Supplementary Fig. S2a and S2c) and NMDAR-mediated EPSC (EPSCNMDA; Supplementary Fig. S2b and S2d). Furthermore, transfection of a scrambled form of NOX2 shRNA had no effect on LTD (scrNOX2 shRNA: 54.9 ± 7.3% of baseline; untransfected cells: 49.3 ± 5.2% of baseline, p = 0.541, Supplementary Fig. S2e). Therefore, our data strongly suggest that ROS production through postsynaptic NOX2 is involved in LTD regulation.
The function of NOX1/NOX2 is principally regulated through multi-site phosphorylation of the p47phox subunit of the complex by PKC isoforms, including the atypical PKCζ27, 28. Given the requirement for NOX2 in LTD, we hypothesized that phosphorylation of p47phox would also be required. To test this, we first utilized biolistic transfection of p47phox shRNA in CA1 neurons of organotypic hippocampal slices to knock down the expression of p47phox (Supplementary Fig. S3). Transfection of p47phox shRNA had no significant effect on basal EPSCAMPA compared with untransfected neighbouring cells (untransfected: 145.0 ± 15.1 pA, transfected: 162.1 ± 15.1 pA, p = 0.594, Fig. 3a). Consistent with our hypothesis, p47phox shRNA transfection significantly impaired LTD expression when compared to untransfected cells (p47phox shRNA-transfected cells: 87.2 ± 11.5% of baseline; untransfected cells: 50.6 ± 7.1% of baseline, p = 0.0145, Fig. 3b).
Fig. 3: P47phox and PKCζ are necessary for LTD.
Since phosphorylation of the p47phox subunit by the atypical PKCζ leads to superoxide generation16, 27, we therefore used PKCζ shRNA (Supplementary Fig. S4) to examine whether PKCζ is also required for LTD. We found that transfection of PKCζ shRNA had no significant effect on basal EPSCAMPA (untransfected: 241.0 ± 33.2 pA, transfected 238.9 ± 22.6 pA, p = 0.957, Fig. 3c) but significantly impaired LTD (PKCζ shRNA: 90.6 ± 4.4% of baseline; untransfected cells: 50.4 ± 5.4% of baseline, p = 0.0001, Fig. 3d). Furthermore, the p47phox shRNA-mediated LTD deficit was rescued by co-expression of human p47phox (untransfected cells: 74.8 ± 8.9% of baseline; transfected cells: 69.2 ± 4.5% of baseline, p = 0.583; Fig. 3e).
To substantiate our hypothesis that PKCζ is an active component of LTD signalling, we tested whether LFS of hippocampal slices, which leads to NMDAR activation and LTD induction, regulates PKCζ activity. To do this, we utilized acutely prepared hippocampal slices prepared from 3-week-old rats. Since PKCζ activity depends on the phosphorylation of threonine 410 (pT410)29, 30, we analysed whether LFS phosphorylates PKCζ at pT410. We found that LFS of Schaffer collateral afferents significantly increased phosphorylation of PKCζ at pT410, indicative of PKCζ activation, and this was blocked by AP5 (50 μM), an NMDAR antagonist (Control: 100 ± 16%; LFS: 185 ± 25%, p < 0.05 compared to control; LFS+AP5: 123 ± 21%, p < 0.05 compared to LFS; AP5: 111 ± 21%, Fig. 3f).
Interestingly, long-term potentiation (LTP) was readily inducible in cells expressing p47phox shRNA (p47phox shRNA: 154.2 ± 6.5% of baseline; untransfected cells: 167.1 ± 14.9% of baseline, p = 0.416, Fig. 3g). LTP was also intact in cells expressing PKCζ shRNA (PKCζ shRNA: 147.9 ± 9.4% of baseline; untransfected cells: 160.1 ± 10.0% of baseline, p = 0.391, Fig. 3h), indicating a selective role for p47phox and PKCζ in the LTD form of synaptic plasticity. Together, these data indicate that PKCζ activation downstream of NMDAR activation is a fundamental component of LTD signalling, regulating the activity of NOX2 via p47phox.
Since both p47phox shRNA and PKCζ shRNA transfection resulted in a loss of LTD, it was of interest to determine the specific molecular mechanism surrounding the regulation of p47phox by PKCζ. We generated shRNA-resistant constructs of rat p47phox with differing combinations of site-specific mutations at four residues phosphorylated by PKCζ27. Consistent with our working hypothesis, LTD was blocked in cells co-transfected with p47phox shRNA and an shRNA-resistant mutant form of p47phox with all four residues mutated to alanine, to prevent their phosphorylation (after LFS: 95.1 ± 6.8%, p = 0.725 vs. control input, Fig. 4a). In cells expressing a triple phosphorylation mutant (S/T304/305/316A—residues notated as per rat p47phox), LTD was also impaired (after LFS: 88.0 ± 5.5%, p = 0.087 vs. control input, Fig. 4b). In comparison, LTD was readily inducible in cells transfected with a double S/T304/305A mutant (after LFS: 57.2 ± 4.1%, p = 0.000117 vs. control input, Fig. 4c). Notably, we found that LTD was also impaired in cells expressing the S316A mutant form of p47phox (after LFS: 98 ± 22.8%, p = 0.73 vs. control input, Fig. 4d) but the expression of S329A mutant form of p47phox had no effect on LTD induction (after LFS: 63.1 ± 7.0%, p = 0.004 vs. control input, Fig. 4e). Finally, we tested whether phosphorylation of p47phox at S316 is sufficient to induce synapse weakening, through paired recordings from neurons transfected with a S316 pseudo-phosphorylated form of p47phox (S316D) and neighbouring untransfected neurons. Our data showed significantly reduced AMPAR-mediated currents in S316D-transfected neurons (untransfected: 132.4 ± 12.9 pA vs. S316D transfected: 94.1 ± 9.4 pA, p = 0.023, Fig. 4f), indicating that phosphorylation at this residue is sufficient to reduce AMPAR-mediated synaptic transmission. This effect appears specific to AMPARs, as NMDAR-mediated currents were unchanged between the two cell types (untransfected: 131.1 ± 11.7 pA vs. S316D transfected: 118.7 ± 13.7 pA, p = 0.495, Fig. 4f). Taken together, these data suggest that phosphorylation at S316 of p47phox, likely by PKCζ, is a key regulator of a signalling cascade that governs LTD induction and synapse weakening.
Fig. 4: Phosphorylation of p47phox at S316 is required for LTD.
The notion that ROS production is a key element of synaptic plasticity has been well established11,12,13, 20, 31. However, the precise source of ROS and mechanisms by which it is regulated during synaptic plasticity have not been fully elucidated. Identification of these specific signalling pathways is necessary for our full understanding of its physiological and pathophysiological implications in synaptic function.
In the present study, we have shown that postsynaptic NOX2-mediated superoxide production, via PKCζ-mediated phosphorylation of p47phox at the serine 316 residue (pS316 p47phox), is pivotal for LTD expression and weakening of AMPAR-mediated synaptic transmission. Importantly, few studies have directly addressed the source of ROS in the context of synaptic plasticity. Using our selective knockdown approach, in which we have specifically silenced or inhibited postsynaptic ROS production, we have now shown that postsynaptic NOX2 is a necessary hub for ROS production associated with LTD. Several research groups have shown a similar requirement for ROS during LTD or synapse weakening9, 10, 12, 13 whilst evidence from other groups suggests that ROS can regulate LTP12, 31, 32. Quite how or why ROS can regulate both forms of synaptic plasticity is unclear, but it is possible that different ROS production sources and mechanisms may underpin different forms of synaptic plasticity.
NMDAR activation leads to PKCζ-mediated phosphorylation of the p47phox subunit, which is a critical activator signal for NOX2 mediated ROS production15, 16. In the present study, we show that low-frequency electrical stimulation of hippocampal slices, which induces an NMDAR-dependent form of LTD, also leads to the activation of PKCζ. This effect was blocked by D-AP5, indicative of synaptic NMDAR-dependent activation of PKCζ that is associated with LTD induction. Importantly, through postsynaptic knockdown of PKCζ expression, we reveal an LTD-specific requirement for PKCζ with no observable contribution to LTP expression, consistent with other synapse weakening signals shown in previous studies5, 6, 33,34,35,36. Finally, our data show that constitutive phosphorylation of the PKCζ substrate, p47phox, at S316 is sufficient to induce weakening of AMPAR-mediated synaptic transmission, even in the absence of upstream activator signals. This single phosphorylation event, which induces NOX2 activation and ROS production, is therefore both necessary and sufficient for synapse weakening. It is not clear how postsynaptic ROS production is itself involved in the mechanisms of LTD signalling. One possibility is that postsynaptic ROS activates Bax protein to stimulate cytochrome c release from mitochondria37. Cytochrome c release induces caspase-3 activation38, which in turn can affect synapse weakening signal cascades involving Akt-1 and GSK-3β4, 39. This possibility is supported by the observation that Bax is itself required for LTD signalling33.
A growing list of molecules, including caspases, GSK-3β and tau, are now known to be involved in AMPAR endocytosis and LTD5, 6, 33, 34. Collectively, these molecules form what we have termed the synapse weakening pathway, encompassing molecules associated with apoptosis and synapse elimination in both physiological and pathophysiological circumstances4, 36, 40. It has been postulated that the balance between synapse weakening (caspase-3, GSK-3β and tau) and strengthening pathways (phosphoinositide-3 kinase and Akt-1) is critical to determine the direction of LTP and LTD and long-term fate of synapses4,5,6, 35. Indeed, models of neurodegenerative pathologies such as AD exhibit AMPAR endocytosis and facilitated LTD induction, concomitant with the inhibition of LTP in the hippocampus2, 3, 39, 41, 42. Aberrant activation of synapse weakening signals is therefore believed to be a central underlying molecular mechanism in the pathology and cognitive decline of numerous neurodegenerative diseases4, 40, 43, 44. Our results suggest that ROS, via a specific production mechanism, now form a part of this critical synapse weakening signalling cascade. Addressing whether aberrant LTD-like and/or AMPAR-mediated synapse weakening can be seen in human forms of neurodegenerative disease remains a key challenge to translating these findings into viable therapeutic targets.
All procedures involving animals were carried out in accordance with the UK Animals (Scientific Procedures) Act, 1986. Male Wistar rats (Charles River, UK) were used to prepare organotypic (6–8 day-old rats) and acute hippocampal slices (2–4-week-old rats). Older rats were housed four or five per cage and allowed access to water and food ad libitum. The cages were maintained at a constant temperature (23 ± 1 °C) and relative humidity (60 ± 10%) under a 12-h light/dark cycle (lights on from 07:30 to 19:30).
Rats were killed by cervical dislocation and decapitation. Following this, the brain was rapidly removed and placed into ice-cold artificial cerebrospinal fluid (aCSF; continuously bubbled with 95% O2/5% CO2) containing 124 mM NaCl, 3 mM KCl, 26 mM NaHCO3, 1.25 mM NaH2PO4, 2 mM CaCl2, 1 mM MgSO4, and 10 mM d-glucose. Hippocampi were extracted and transverse hippocampal slices (400 µm thickness) were cut using a McIlwain tissue chopper. Following manual separation, the slices were then submerged in aCSF for a minimum of 1 h before experiments commenced.
Acute slices were placed in a recording chamber and perfused with warmed (28–29 °C) and carbogenated aCSF at 2 ml/min. Two independent stimulating electrodes were placed separately in the Schaffer collateral–CA1 input (test pathway) and subiculum–CA1 input (control pathway). For whole-cell patch experiments, 20 μM picrotoxin was included in the aCSF and CA1 neurons were blind-patched using a 4–6 MΩ borosilicate glass pipette containing 130 mM CsMeSO4, 8 mM NaCl, 4 mM Mg-ATP, 0.3 mM Na-GTP, 0.5 mM EGTA, 10 mM HEPES, 6 mM QX-314, pH 7.2–7.3 and 280–290 mOsm/kg. For field excitatory postsynaptic potential (fEPSP) recordings, a glass pipette containing 3 M NaCl was placed in the stratum radiatum region of the CA1. LTD experiments during whole-cell patch were carried out as for cultured hippocampal slices (see below), except a 20-min baseline was used. For LTD of fEPSPs, a 30-min stable baseline at 70% of maximum stimulation intensity was followed by LFS, consisting of 900 pulses at 1 Hz, of the test pathway. This was followed by 60 min of post-conditioning recording. In some cases, slices were removed immediately after LFS for western blotting processing.
Hippocampal slice cultures were prepared from 6–8-day-old male Wistar rats. Hippocampal slices (350 μm) were cut using a McIlwain tissue chopper and cultured on semipermeable membrane inserts (Millipore Corporation, Bedford, MA, USA) in a six-well plate containing culture medium (78.8% minimum essential medium, 20% heat-inactivated horse serum, 30 mM HEPES, 26 mM d-glucose, 5.8 mM NaHCO3, 2 mM CaCl2, 2 mM MgSO4, 70 μM ascorbic acid, 1 μg/ml insulin, pH adjusted to 7.3 and 320–330 mOsm). Slices were cultured for 6–8 days in vitro (DIV) with a change of medium every 2 days, without antibiotics. Neurons were transfected using a biolistic gene gun (Helios Gene-gun system, Bio Rad, USA) at DIV3–4 (100 μg DNA; 90% of the construct to test; 10% pEGFP-C1). Electrophysiological recordings were performed 3–4 days after transfection.
For whole-cell patch recordings, cultured slices were perfused with a warmed (28–29 °C) recording solution (119 mM NaCl, 2.5 mM KCl, 26 mM NaHCO3, 1 mM NaH2PO4, 4 mM MgCl2, 11 mM d-glucose, 4 mM CaCl2, 10 μM 2-chloroadenosine and 20 μM picrotoxin). The recording solution was continuously bubbled with 95% O2/5% CO2 at source. The usual flow rate was 2 ml/min. In most recordings, two independent stimulating electrodes were placed separately in the Schaffer collateral–CA1 input and subiculum–CA1 input. Recordings were made from pyramidal neurons in the CA1 region, using glass pipettes containing CsMeSO4 internal solution (as above) and neurons voltage clamped at −70 mV unless otherwise stated. To induce LTD, a 10-min baseline was followed by 1 Hz stimulation (200 stimuli) with recorded neurons voltage clamped at −40 mV. To induce LTP, a 5-min baseline was followed by a 2 Hz stimulation (200 stimuli) with recorded neurons voltage clamped at 0 mV. For quantification and comparisons between groups/inputs, the peak EPSC amplitude of the test input (relative to baseline) was averaged 15–20 min (cultured slice) or 20–25 min (acute slice) after conditioning was applied. EPSCAMPA was measured as the peak EPSC amplitude at a holding potential of −70 mV. EPSCNMDA was measured as the peak EPSC amplitude 90–100 ms after stimulus, at a holding potential of +40 mV.
The following drugs were dissolved in internal recording solution (for postsynaptic infusion) or aCSF (for bath perfusion) at concentrations based on previous studies: AEBSF (20 μM45; Sigma-Aldrich, MO, USA), apocynin (100 μM46; Abcam, Cambridge, UK), SOD (300 units/ml47; Sigma-Aldrich, MO, USA), catalase (300 units/ml48; Sigma-Aldrich, MO, USA), ML-171 (3 μM49; Tocris, Oxford, UK) and AP5 (Hello Bio, Bristol, UK). Primary antibodies used for western blotting were pT410 PKCζ (Cell Signaling, MA, USA), total PKCζ (Santa Cruz, Texas, USA) and total β-actin (Abcam, Cambridge, UK).
Constructs for shRNA knockdown of target transcripts were generated using the Block-iT™ pENTR/U6 system, as per the manufacturer's instructions (Life Technologies, UK). The target sequences for rat NOX1 and NOX2 were GCAACTGTTCATACTCTTTCC and GGTCTTACTTTGAAGTGTTCT, respectively. NOX2 scrambled shRNA sequence was GGTAGTTACTCGTTAGTTTCT. PKCζ shRNA target sequence was GGCCATGAGCATCTCTGTTGT and for p47phox it was GCTCCTACCCTGCTTTAATGT. P47phox-rescue and mutation experiments were performed with co-transfection of the p47phox shRNA construct and a pCMV-SPORT6-p47phox cDNA clone (Source Bioscience, UK). Site-directed mutagenesis to generate S/T304/305A, S/T304/305/316A, S316A and S329A constructs was performed on the pCMV-SPORT6-p47phox construct using QuikChange™ technology, as per the manufacturer's instructions (Agilent Technologies, USA). All generated constructs were sequence verified via Sanger sequencing (Source Bioscience, UK).
Knockdown efficiencies of the generated shRNAs were verified by neuronal single-cell PCR. Briefly, transfected and non-transfected control cells were excised from cultured slices using low-resistance electrodes and then snap frozen. Cells were then subjected to OneStep™ RT-PCR (Qiagen, The Netherlands), as per the manufacturer's instructions, using specific exon-spanning primers: NOX1: sense 5′-AGAGGCTCCAGACCTCCATTT-3′, anti-sense, 5′-CGTGTGGTTGCAAAATGAGCA-3′; NOX2: sense 5′-AGCACTTCACACGGCCATTC-3′, anti-sense 5′-AGAGGTCAGGGTGAAAGGGT-3′; PKCζ: sense 5′-GGACCTCTGTGAGGAAGTGC-3′, anti-sense, 5′-GGATGCTTGGGAAAACGTGG-3′; PKMζ: sense 5′-CCTTCTATTAGATGCCTGCTCTCC-3′, anti-sense 5′-TGAAGGAAGGTCTACACCATCGTTC-3′50; p47phox: sense 5′-CACCTTCATTCGCCACATCG-3′, anti-sense 5′-ATGTCCCTTTCCTGACCAC-3′; Cyclophilin: sense 5′-CCAAGACTGAGTGGCTGGAT-3′, anti-sense 5′- TCCTGCTAGACTTGAAGGGGAA-3′. Amplified PCR products were resolved on a 1% agarose/Midori Green gel and analysed under ultraviolet light.
Sample sizes, indicated by n, are indicated in the figure legends and represent the number of biological replicates. For experiments in acute slices, this reflects the number of individual animals from which the data were obtained. For cultured slices, this reflects the number of individual slices. Samples sizes for electrophysiology experiments were determined through empirical evidence obtained within our laboratory and are consistent with those found in existing literature. Data are expressed as mean ± standard error of the mean (SEM) and analysed using the SigmaPlot software (Systat Software, Chicago, USA). Significance was set at p < 0.05 and unpaired t tests were used to determine the statistical significance of effects vs. control inputs or untransfected cells, where appropriate, and paired t tests were used to compare to baseline values, when necessary.
Citri, A. & Malenka, R. C. Synaptic plasticity: multiple forms, functions, and mechanisms. Neuropsychopharmacology 33, 18–41 (2008).
Hsieh, H. et al. AMPAR removal underlies aβ-induced synaptic depression and dendritic spine loss. Neuron 52, 831–843 (2006).
Shankar, G. M. et al. Amyloid-β protein dimers isolated directly from Alzheimer's brains impair synaptic plasticity and memory. Nat. Med. 14, 837–842 (2008).
Jo, J. et al. Aβ(1-42) inhibition of LTP is mediated by a signaling pathway involving caspase-3, Akt1 and GSK-3β. Nat. Neurosci. 14, 545–547 (2011).
Peineau, S., Taghibiglou, C., Bradley, C. & Liu, L. LTP Inhibits LTD in the hippocampus via regulation of GSK3β. Neuron 53, 703–717 (2007).
Li, Z. et al. Caspase-3 activation via mitochondria is required for long-term depression and AMPA receptor internalization. Cell 141, 859–871 (2010).
Greenlund, L. J., Deckwerth, T. L. & Johnson, E. M. Superoxide dismutase delays neuronal apoptosis: a role for reactive oxygen species in programmed neuronal death. Neuron 14, 303–315 (1995).
Markesbery, W. R. Oxidative stress hypothesis in Alzheimer's disease. Free Radic. Biol. Med. 23, 134–147 (1997).
Wang, Q., Rowan, M. J. & Anwyl, R. Beta-amyloid-mediated inhibition of NMDA receptor-dependent long-term potentiation induction involves activation of microglia and stimulation of inducible nitric oxide synthase and superoxide. J. Neurosci. 24, 6049–6056 (2004).
Ma, T. et al. Amyloid β-induced impairments in hippocampal synaptic plasticity are rescued by decreasing mitochondrial superoxide. J. Neurosci. 31, 5589–5595 (2011).
Massaad, C. A. & Klann, E. Reactive oxygen species in the regulation of synaptic plasticity and memory. Antioxid. Redox Signal. 14, 2013–2054 (2011).
De Pasquale, R., Beckhauser, T. F., Hernandes, M. S. & Giorgetti Britto, L. R. LTP and LTD in the visual cortex require the activation of NOX2. J. Neurosci. 34, 12778–12787 (2014).
Zhang, J. et al. Microglial CR3 activation triggers long-term synaptic depression in the hippocampus via NADPH oxidase. Neuron 82, 195–207 (2014).
Bindokas, V. P., Jordan, J., Lee, C. C. & Miller, R. J. Superoxide production in rat hippocampal neurons: selective imaging with hydroethidine. J. Neurosci. 16, 1324–1336 (1996).
Brennan, A. M. et al. NADPH oxidase is the primary source of superoxide induced by NMDA receptor activation. Nat. Neurosci. 12, 857–863 (2009).
Brennan-Minnella, A. M., Shen, Y., El-Benna, J. & Swanson, R. A. Phosphoinositide 3-kinase couples NMDA receptors to superoxide release in excitotoxic neuronal death. Cell Death Dis. 4, e580 (2013).
McCord, J. M., Keele, B. B. & Fridovich, I. An enzyme-based theory of obligate anaerobiosis: the physiological function of superoxide dismutase. Proc. Natl Acad. Sci. USA 68, 1024–1027 (1971).
Dudek, S. M. & Bear, M. F. Homosynaptic long-term depression in area CA1 of hippocampus and effects of N-methyl-D-aspartate receptor blockade. Proc. Natl Acad. Sci. USA 89, 4363–4367 (1992).
Rosenbaum, D. M., Kalberg, J. & Kessler, J. A. Superoxide dismutase ameliorates neuronal death from hypoxia in culture. Stroke 25, 857–862 (1994). discussion 862–3.
Kamsler, A. & Segal, M. Paradoxical actions of hydrogen peroxide on long-term potentiation in transgenic superoxide dismutase-1 mice. J. Neurosci. 23, 10359–10367 (2003).
Tejada-Simon, M. V. et al. Synaptic localization of a functional NADPH oxidase in the mouse hippocampus. Mol. Cell. Neurosci. 29, 97–106 (2005).
Ueno, N., Takeya, R., Miyano, K., Kikuchi, H. & Sumimoto, H. The NADPH oxidase Nox3 constitutively produces superoxide in a p22phox-dependent manner: its regulation by oxidase organizers and activators. J. Biol. Chem. 280, 23328–23339 (2005).
Martyn, K. D. et al. Functional analysis of Nox4 reveals unique characteristics compared to other NADPH oxidases. Cell. Signal. 18, 69–82 (2006).
Diatchuk, V., Lotan, O., Koshkin, V., Wikstroem, P. & Pick, E. Inhibition of NADPH oxidase activation by 4-(2-aminoethyl)-benzenesulfonyl fluoride and related compounds. J. Biol. Chem. 272, 13292–13301 (1997).
Gianni, D. et al. Optimization and Characterization of an Inhibitor for NADPH Oxidase 1 (NOX-1). Probe Reports from the NIH Molcular Libraries Program [Internet] (National Center for Biotechnology Information (US), Bethesda, MD, 2010).
Jackman, K. A. et al. Reduction of cerebral infarct volume by apocynin requires pretreatment and is absent in Nox2-deficient mice. Br. J. Pharmacol. 156, 680–688 (2009).
Fontayne, A., Dang, P. M.-C., Gougerot-Pocidalo, M. A. & El-Benna, J. Phosphorylation of p47phox sites by PKC alpha, beta II, delta, and zeta: effect on binding to p22phox and on NADPH oxidase activation. Biochemistry 41, 7743–7750 (2002).
Bedard, K. & Krause, K.-H. The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology. Physiol. Rev. 87, 245–313 (2007).
Chou, M. M. et al. Regulation of protein kinase C zeta by PI 3-kinase and PDK-1. Curr. Biol. 8, 1069–1077 (1998).
Hirai, T. & Chida, K. Protein kinase Czeta (PKCzeta): activation mechanisms and cellular functions. J. Biol. Chem. 133, 1–7 (2003).
Kishida, K. T. et al. Synaptic plasticity deficits and mild memory impairments in mouse models of chronic granulomatous disease. Mol. Cell. Biol. 26, 5908–5920 (2006).
Thiels, E. et al. Impairment of long-term potentiation and associative memory in mice that overexpress extracellular superoxide dismutase. J. Neurosci. 20, 7631–7639 (2000).
Jiao, S. & Li, Z. Nonapoptotic function of BAD and BAX in long-term depression of synaptic transmission. Neuron 70, 758–772 (2011).
Kimura, T. et al. Microtubule-associated protein tau is essential for long-term depression in the hippocampus. Philos. Trans. R. Soc. Lond. B. Biol. Sci. 369, 20130144–20130144 (2014).
Regan, P. L. et al. Tau phosphorylation at serine 396 residue is required for hippocampal LTD. J. Neurosci. 35, 4804–4812 (2015).
Yi, J.-H. et al. Glucocorticoids activate a synapse weakening pathway culminating in tau phosphorylation in the hippocampus. Pharmacol. Res. 121, 42–51 (2017).
Gomez-Lazaro, M. et al. Reactive oxygen species and p38 mitogen-activated protein kinase activate Bax to induce mitochondrial cytochrome c release and apoptosis in response to malonate. Mol. Pharmacol. 71, 736–743 (2007).
Cai, J., Yang, J. & Jones, D. P. Mitochondrial control of apoptosis: the role of cytochrome c. Biochim. Biophys. Acta 1366, 139–149 (1998).
Regan, P. L., Whitcomb, D. J. & Cho, K. Physiological and pathophysiological implications of synaptic tau. Neuroscientist https://doi.org/10.1177/1073858416633439 (2016).
Shipton, O. A. et al. Tau protein is required for amyloid β-induced impairment of hippocampal long-term potentiation. J. Neurosci. 31, 1688–1692 (2011).
Kim, J. H., Anwyl, R., Suh, Y. H., Djamgoz, M. B. & Rowan, M. J. Use-dependent effects of amyloidogenic fragments of (beta)-amyloid precursor protein on synaptic plasticity in rat hippocampus in vivo. J. Neurosci. 21, 1327–1333 (2001).
Shankar, G. M. et al. Natural oligomers of the Alzheimer amyloid-beta protein induce reversible synapse loss by modulating an NMDA-type glutamate receptor-dependent signaling pathway. J. Neurosci. 27, 2866–2875 (2007).
Selkoe, D. J. Alzheimer's disease is a synaptic failure. Science 298, 789–791 (2002).
Day, M. et al. Selective elimination of glutamatergic synapses on striatopallidal neurons in Parkinson disease models. Nat. Neurosci. 9, 251–259 (2006).
Kovac, S. et al. Nrf2 regulates ROS production by mitochondria and NADPH oxidase. Biochim. Biophys. Acta 1850, 794–801 (2015).
Lull, M. E., Levesque, S., Surace, M. J. & Block, M. L. Chronic apocynin treatment attenuates beta amyloid plaque size and microglial number in hAPP(751)(SL) mice. PLoS ONE 6, e20153 (2011).
Preston, J. E., Hipkiss, A. R., Himsworth, D. T., Romero, I. A. & Abbott, J. N. Toxic effects of beta-amyloid(25-35) on immortalised rat brain endothelial cell: protection by carnosine, homocarnosine and beta-alanine. Neurosci. Lett. 242, 105–108 (1998).
Yu, A. C., Gregory, G. A. & Chan, P. H. Hypoxia-induced dysfunctions and injury of astrocytes in primary cell cultures. J. Cereb. Blood Flow Metab. 9, 20–28 (1989).
Cetinbas, N. et al. Loss of the tumor suppressor Hace1 leads to ROS-dependent glutamine addiction. Oncogene 34, 4005–4010 (2015).
Marchand, F. et al. Specific involvement of atypical PKCζ/PKMζ in spinal persistent nociceptive processing following peripheral inflammation in rat. Mol. Pain 7, 86 (2011).
This study was supported by a UK Wellcome Trust collaborative grant to K.C. and the BBSRC to K.C. and D.J.W. and the Korea-UK Alzheimer's disease Research Consortium grant to D.H.K. and J.H.Y. (Korean Ministry of Health and Welfare). K.C. was supported by the UK Dementia Research Institute.
The study was conceived and designed by K.C. Electrophysiological studies were conducted by D.H.K., J.H.Y., S.C.K. and P.R., constructs were produced by T.P. and biochemical assays were conducted by D.H.K. and J.H.Y. The manuscript was written by P.R., T.P., J.H.Y., D.J.W. and K.C.
Correspondence to Philip Regan or Kwangwook Cho.


Vortrag: ACM International Conference on Multimedia Retrieval (ICMR 2015), Shanghai, China; 23.06.2015 - 26.06.2015; in:"ACM International Conference on Multimedia Retrieval (ICMR 2015)", (2015), 8 S.
A significant part of publicly available photos on the Internet depicts a variety of different social events. In order to organize this steadily growing media content and to make it easily accessible, novel indexing methods are required. Essential research questions in this context concern the efficient detection (clustering), classification, and retrieval of social events in large media collections. In this paper we explore two aspects of social events mining. First, the initial clustering of a given photo collection into single events and, second, the retrieval of relevant social events based on user queries. For both aspects we employ commonly available metadata information, such as user, time, GPS data, and user-generated textual descriptions. Performed evaluations in the context of social event detection demonstrate the strong generalization ability of our approach and the potential of contextual data such as time, user, and location. Experiments with social event retrieval clearly indicate the open challenge of mapping between previously detected event clusters and heterogeneous user queries.


Toronto-area estate lawyer Charles Ticker is scheduled to speak on the importance of making informed legal decisions as you age at a panel discussion next month.
The event, taking place at Congregation Darchei Noam, Toronto's Reconstructionist Synagogue, is called A Symposium on Liberal Jewish Responses to Aging.
Running from 2 to 6 p.m., the symposium aims to offer participants a chance to learn how medicine, Canadian law and liberal halachic perspectives can help inform decisions regarding health, finances and relationships. Preparing in advance for aging years and opening doors to important conversations to avoid family conflict will also be part of the discussion.
Ticker is slated to speak on the topic of power of attorneys. Other presenters are Dr. Sandy Buchman and Rabbi Michael Stroh.
Registration is now open for the event, taking place Dec. 2. All participants will choose two workshops and receive and resource binder. More information can be found by calling 416-638-4783 ext. 24.


text:
Terminally ill patient welcomes 'humane' future for dying Victorians
Victorian economy to bounce back fastest: Report
Zero tolerance vital: Racism in any form just isn't cricket
Dennis Atkins: Scott Morrison's antics show Australia is going to the polls in 2021
Michael Pascoe: From QAnon to climate denial, Australia has a 'Ratbag Government'
Capitol insurrection: American democracy is bruised and sore, so what happens next?
News State Victoria News Terminally ill patient welcomes 'humane' future for dying Victorians
5:24pm, Nov 29, 2017 Updated: 7:15pm, Nov 29
Jen and Ken Barnes say the voluntary assisted dying laws give them comfort. Photo: ABC
Victorians will have the right to request a lethal drug to end their lives from mid-2019, but the new legislation will likely come too late for Jen Barnes.
After more than 100 hours of debate across both houses of Parliament and two demanding all-night sittings, Lower House MPs on Wednesday ratified the amended bills, which will now be sent for Royal assent.
Premier Daniel Andrews, who came to support euthanasia after the death of his father last year, said it was an historic day.
Daniel Andrews (right) Jill Hennessy (centre), and Victorian Attorney-General Martin Pakula (left) congratulated MPs after hours of debate.
"This is a day of reform, a day of compassion, a day of giving control to those who are terminally ill," he said.
Barnes has just been told the blob-like tumour on her brain is growing, inoperable and will ultimately rob her of life.
But with the little time she has left, she and her husband Ken have, for months, been regularly making the four-hour round trip to the Victorian Parliament from their home on the coast to watch politicians debate the new legislation.
"I just want to know that it's there, the option is there," she told 7.30.
"You don't know until you're actually there and getting close to the moment, you don't know how you're going to react. You don't know if you're going to say, 'No, no, I want that next day,' or whatever, [but] it's the comfort of it."
Ms Barnes was overjoyed when the legislation cleared its final hurdle.
"It's unlikely that I would see it come through for me because I'm not likely to be here in 18 months' time," she said.
"But for the future, I think it's the right way to go.
"I mean they always said that it's only going to affect a small number of people but for those people I think it's important that it's there."
Doctors still divided over legislation
While some patients rejoiced over the legislation passing, doctors met the news with unease.
Associate professor Mark Boughey, director of palliative medicine at Melbourne's St Vincent's Hospital, said he still had major concerns about what voluntary assisted dying would mean for patients.
"I think anyone that's facing the end of their life is vulnerable," he told 7.30.
When you consider more vulnerable people, those who are coming from non-English speaking backgrounds or Aboriginal or Torres Strait Island backgrounds, I think again there will be a whole raft of concerns that will be difficult for the legislation to be explained and understood by those people."
He won't have to go against his beliefs because earlier this year his employer, Catholic organisation St Vincent's Health, announced it would ban its staff from taking part in voluntary assisted dying if it was legalised.
But Dr Boughey said it was still a bad day for patient choice.
"My biggest concern is that people will see the progression from illness and disease treatment, the next step will be assisted dying," he said.
"They won't get the benefit of engagement and referral to palliative care.
"My other concern is that they will feel a burden within their home and their family and therefore that's an alternative that they'll feel pressured to consider."
'None of us get out of this life alive'
Ms Barnes believes the concerns about coercion and pressure are unfounded, given the dozens of safeguards in the legislation.
Her husband is also on board.
Images of Jen Barnes' brain tumor.
"There's no argument, we're all on the same page," Mr Barnes said. "Death is death, it's part of life, it's the dying bit that's the problem.
"This is the most practical and humane decision for all of us."
As he sits with his wife back at their Inverloch home, the pair are flicking through funeral brochures over a cup of tea.
"So this is where we'll both finish up. You go there first and let me know what it's like," Mr Barnes joked.
Ms Barnes isn't sure she would take up voluntary assisted dying even if she does live until the law is implemented, but she's already decided where she wants to end up.
"I'm going to a place called Mount Elephant, and you get buried standing up and hawks fly around there and they plant a native tree there," she said.
"It's the cheapest funeral you can have and I thought, 'That'll do me'."
-ABC
AFL: Sun shines on Carlton as Garlett settles in to revive his career

redpajama_set_name: RedPajamaCommonCrawl


text:
Harmon's Histories: Hammond Arcade rises once more to celebrate Art Deco past
Missoula Current Published: January 16, 2023
Missoulian March 23,1934
"Live, by remote control," direct from the studio in the Hammond Arcade, it's "Paul Keith and His Arcade Orchestra!"
That's right, Missoula's Hammond Arcade had its own orchestra, appearing on its own radio show (the Hammond Radio Review) for the downtown building's grand opening in 1934.
The broadcast took place Friday and Saturday, March 23 and 24 from 3 to 5 p.m., and again from 7 to 9 p.m. on KGVO radio!
The new Hammond Arcade rose from the ashes of the original "Hammond block," a four-story brick building constructed in 1890.
The original block, destroyed by fire on October 9, 1932, included 10 businesses on the ground level, plus 25 apartments and offices above.
The state highway commission had offices on the second floor. The division engineer lost his personal diary books and "some records, maps and equipment."
Dr. F.G. Dratz, a local dentist, had his office on the second floor as well. He was away, fishing at Placid Lake, at the time. When he returned, he was able to salvage his records, stored in a steel cabinet.
Other tenants like the Postal Telegraph Company and a hamburger shop scrambled to find temporary business quarters, as did the largest tenant, the Missoula Drug Company.
Although damage was estimated at $200,000 ($4,350,510 in today's dollars), Sid Coffee and his partners, Walter L. Pope and Oakley E. Coffee, owners of the 100 block of South Higgins, did not hesitate in announcing immediate plans to rebuild.
They were true to their pledge. A year and a half later, the reconstruction was complete and the grand opening was announced.
Hundreds See New Building March 24, 1934
The new building, while not the multi-story structure it replaced, was celebrated as a "new, fireproof structure, built of reinforced concrete. It covered "more than 11,000 square feet" and included "two city blocks of glass ... the only (such building) of its kind in Missoula."
The interior was finished with Philippine mahogany and "exquisite (tile flooring) that harmonized with the interior decorations." The central heating system, located in the basement, eliminated the need for unsightly radiators.
The stoker-fired boiler was "believed to be one of the most up-to-date heating systems in the Northwest."
The plumbing and heating contractor, Pope & Hintz, was also proud of the modern women's restroom "located on the main floor and open to the public at all hours."
New Design Appreciated 1-7-34
Flash forward to 2023, and we find the current owner of the block announcing plans to remodel again, this time honoring the 1934 Art Deco design (which had been altered considerably over the years).
Nick Caras told the Missoula Current, "With these improvements, the building will be one step closer to its original design and another step closer to being more accessible and approachable to the Missoula downtown community."
Jim Harmon is a longtime Missoula news broadcaster, now retired, who writes a weekly history column for Missoula Current. You can contact Jim at fuzzyfossil187@gmail.com. His best-selling book, "The Sneakin'est Man That Ever Was," a collection of 46 vignettes of Western Montana history, is available at harmonshistories.com.
Filed Under: feature, Hammond Arcade, Missoula history, Montana Today, Radio RSS
Categories: Up in the 406


text:
Fracking Industry Puts Communities at Risk
Robert F. Kennedy, Jr.
Oct. 26, 2011 12:53PM EST Fracking
Superb investigative journalism by the New York Times has brought the paper under attack by the natural gas industry. That campaign of intimidation and obfuscation has been orchestrated by top shelf players like Exxon and Chesapeake aligned with the industry's worst bottom feeders. This coalition has launched an impressive propaganda effort carried by slick PR firms, industry funded front groups and a predictable cabal of right wing industry toadies from cable TV and talk radio. In pitting itself against public disclosure and reasonable regulation, the natural gas industry is once again proving that it is its own worst enemy.
I confess to being an early optimist on natural gas. In July of 2009, I wrote a widely circulated op-ed for the Financial Times predicting that newly accessible deposits of natural gas had the potential to rapidly relieve our country of its deadly addiction to Appalachian coal and end forever catastrophically destructive mountaintop removal mining. At that time, government and industry geologists were predicting that new methods of fracturing gas rich shale beds had provided access to an astounding 2000-5000 trillion cubic feet of natural gas in the lower 48—enough, they claimed to power our country for a century.
These rich reserves might have allowed America to mothball or throttle back our 336 gigawatts of mainly antiquated and inefficient coal fired electric plants replacing them with underutilized capacity from existing gas generation plants. That transition could reduce U.S. mercury emissions by 20-25 percent, dramatically cut deadly particulate matter and the pollutants that cause acid rain and slash America's grid based CO2 by an astonishing 20 percent—literally overnight! Gas could have been a natural companion for wind and solar energy with its capacity to transform variable power into base load, and could have been a critical bridge fuel to the new energy economy rooted in America's abundant renewables.
American sourced natural gas might also have helped free us from our debilitating reliance on foreign oil now costing our country so dearly in blood, national security, energy independence, global leadership, moral authority and treasure amounting to $700 billion per year—the total cost to our country of annual oil imports—in addition to two pricey wars that are currently running tabs of $2 billion per week.
My caveat was that the natural gas industry and government regulators needed to act responsibly to protect the environment, safeguard communities from irresponsible practices, and to candidly inform the public about the true risks and benefits of shale extraction gas.
The opposite has happened.
The industry's worst actors have successfully battled reasonable regulation, stifled public disclosure while bending compliant government regulators to engineer exceptions to existing environmental rules. Captive agencies and political leaders have obligingly reduced already meager enforcement resources and helped propagate the industry's deceptive economic projections. As a result, public skepticism toward the industry and its government regulators is at a record high. With an army of more than 40,000 highly motivated anti-fracking activists in New York alone, popular mistrust of the industry is presenting a daunting impediment to its expansion.
I sit on the New York State Governor Andrew Cuomo's High Volume Hydraulic Fracturing Advisory Panel. I and the other panelists are charged with developing recommendations to the Commissioner regarding rules that will hopefully safeguard New Yorkers from the kind of calamities caused by the natural gas industry to communities just across our border with Pennsylvania. We spend much of our time sorting truth from the web of myths spun about fracking by fast talking landsmen, smarmy CEOs and federal regulators.
Recent studies have raised doubts about many of the industry's fundamental presumptions;
• For example, releases of methane, a far more potent greenhouse gas, may counterbalance virtually all the benefits of CO2 reductions projected to result from substituting gas power for coal. [Robert W. Howarth, Renee Santoro, Anthony Ingraffea, Methane and the greenhouse-gas footprint of natural gas from shale formations, Climatic Change (2011); Wigley T. (2011) Coal to Gas: The Influence of Methane Leakage. Climate Change Letters. DOI 10.1007/s10584-011-0217-3].
• The human health impacts of gas extraction on local communities may rival those associated with coal. A new study by Centers for Disease Control finds that breast cancer rates have dropped in every county in Texas, but have increased in the six counties with the heaviest natural gas air emissions.
• The U.S. Geological Survey just slashed its estimate on the amount of gas in the Marcellus Shale by 80 percent, raising doubts about all the industry's positive economic projections about jobs, royalties and revenues. Industry based those projections on resource estimates that the federal government has now jettisoned.
• Meanwhile local communities are finding the costs of irresponsible drilling to be ruinous. Contaminated well water, poisoned air, nuisance noise and dust, diminished property values and collapsing quality of life are often the predictable collateral damage of gas shale development in the rural towns of the east. [Barth. The Unanswered Questions About the Economic Impact of Gas Drilling in the Marcellus Shale: Don't Jump to Conclusions. March 2010. Accessed 8/10/11; Christopherson & Rightor. How Should We Think About the Economic Consequences of Shale Gas Drilling? May 2011. Accessed 8/10/11;Stephen G. Osborn, Avner Vengosh, et al., Methane Contamination of drinking water accompanying gas wells drilling and hydraulic fracturing, PNAS Early Edition, April 14, 2011; Riverkeeper, Fractured Communities (Sept. 2010)].
• In a devastating admission, the industry now acknowledges that it absolutely cannot afford to pay localities the costs of roads damaged from the thousands of truck trips per wellhead, leaving those ruinous costs to local taxpayers, many of whom will see no benefits from the shale boom, but only declines in their quality of life.
• With several notable exceptions, like Southwest Energy, the industry has demonstrated a disturbing fervor for secrecy while advocating regulatory policies that favor the most irresponsible practices and the worst actors.
The shale gas industry's campaign against The Times illustrates the difficulty in getting solid information upon which to base a regulatory scheme. The Times is doing an unusually rigorous job at covering this extremely important and complex issue. The paper's ongoing series on natural gas drilling is one of the strongest pieces of investigative journalism this year from any news venue. Thankfully, The Times is covering this extremely important topic with rigor and balance. But it is also going the extra mile in the level of documentation it provides to bolster its stories, a move that raises the bar on public service journalism.
In an era when few papers or news outlets are still willing to take on very powerful interests, The Times has pursued very difficult questions about one of our country's richest and most aggressive industries. At a time when accessing documents through open records requests faces an obstacle course of daunting roadblocks, the series has spent nearly a year using these flawed tools to collect and publish an extraordinary trove of original documentation. Archives published by The Times include thousands of pages obtained through leaks and/or public records requests. The Times reporters provide page-by-page annotations explaining the documents so that the reader can sift through them in guided fashion.
Among the revelations uncovered by The Times' admirable reporting:
• Sewage treatment plants in the Marcellus region have been accepting millions of gallons of natural gas industry wastewater that carry significant levels of radioactive elements and other pollutants that they are incapable of treating.
• An EPA study published by The Times shows receiving rivers and streams into which these plants discharge are unable to consistently dilute this kind of highly toxic effluent.
• Most of the state's drinking water intakes, streams and rivers have not been tested for radioactivity for years—since long before the drilling boom began.
• Industry is routinely making inflated claims about how much of its wastewater it is actually recycling.
• EPA, caving to industry lobbyists and high level political interference reminiscent of the Bush/Cheney era, has narrowed the scope of its national study on hydrofracking despite vocal protests from agency scientists. The EPA had, for example, planned to study in detail the effect on rivers of sending radioactive wastewater through sewage plants, but dropped these plans during the phase when White House-level review was conducted.
• Similar studies in the past had been narrowed by industry pressure, leading to widespread exemptions for the oil and gas industry from environmental laws.
• The Times revealed an ongoing and red hot debate within the EPA about whether the agency should force Pennsylvania to handle its drilling waste more carefully and strengthen that state's notoriously lax regulations and anemic enforcement.
• The Times investigation also explodes the industry's decade old mantra that a "there is not a single documented case of drinking water being contaminated by fracking." The Times investigation of EPA archives exposes this claim as demonstrably false.
A second round of New York Times stories showed that within the natural gas industry and among federal energy officials, there were serious and disturbing reservations about the economic prospects of shale gas:
• Government and industry officials made sure that all of their reservations were discussed privately and never revealed to the American public. Internal commentary by these officials is striking because it contrasts so sharply with the excited public rhetoric from the same agencies, lawmakers, industry officials and energy experts about shale gas.
• Many industry experts have reservations over whether the wells produce as much gas as industry is claiming and whether companies may be misleading investors, landowners, and the public about the true costs of shale gas.
• Shale gas wells often dry up faster than companies expected—sometimes several decades faster than predicted.
• Rather than coming clean, the companies downplay how much it costs to keep these wells flowing and overstate how much profit companies can make by these wells.
• Furthermore, only a small percentage of the land in each shale gas field turns out to be highly productive, even at the start. Nevertheless, companies routinely pretend that all of their acreage will be equally promising.
• These emerging issues also sparked private discussion among federal energy experts, who expressed grave concern that their agency's predictions were too heavily influenced by the natural gas industry's over-optimism. The Times found that the EIA was heavily reliant on data provided by companies with shale-gas industry ties.
The science writer for the Knight Ridder Journalism website summed up the significance of The Times' revelations about the industry's ballyhooed economic prospects. "From here, it appears that the Times and [the series principal author] Mr. Urbina are calmly saying we should learn a lesson from the dot-com bubble and the housing bubble, suggesting investors and regulators and gov't planners step with care and not be blinkered by all the money that's pouring in."
The organized attack on The Times and its reputation by well-financed industry spin machines is illustrative of the perils and real challenges facing public service journalism today.
The Times piece has been the target of massive industry blowback. Industry funded front groups like Energy in Depth, an army of slick PR firms, and former regulatory officials like PA DEP Commissioner John Hanger, now on industry's payroll, have artfully manufactured deceptive talking points and posted blogs that are parroted by journalists looking for an industry response to The Times coverage and then emailed as "facts" to the industry's supporters and its indentured servants in Congress.
Ironically, many of the attacks against the series have claimed that the articles were poorly sourced or under-researched. Yet, The Times has not printed a single factual correction. This is certainly an admirable reporting record for a series that has been running in the paper for nearly a year. This is because, despite massive efforts by the industry to find errors, no critic has been able to identify a single fact that The Times actually got wrong. The Times posted thousands of pages of closely annotated original-source documents along with its news articles.
Rarely has a series had such wide-reaching and immediate impact. The New York Times articles have led to major changes in how the industry as well as state and federal regulators are handling one of America's most important energy issues.
Documents uncovered by The Times have already been put to use in litigation by injured parties seeking to force some treatment plants to stop handling the frack wastewater. The Times series has also pressured the EPA to begin a review of treatment plant permits (signaling the agency's possible intent to prohibit plants from discharging treated waste into rivers without comprehensive testing for shale gas contaminants). Healthy skepticism raised by the series has dampened some of the thrilled exuberance among Wall Street bankers ecstatic about the latest gold rush, federal lawmakers in the thrall of industry money, and in hard pressed rural communities seduced by hollow promises of massive royalties, local prosperity and abundant jobs.
As our panel grapples with these complex and difficult problems, we have found that the principal impediment to going forward with recommendations regarding regulations that could allow fracking in our state is a general mistrust of the claims we are hearing from industry and federal regulators. Revelations from The Times series and elsewhere have cast doubt upon all the industry's assurances about fracking and have complicated the task for those of us charged with advising the regulatory agencies on developing rules that could allow the industry to proceed while safeguarding the public interest.
For many of us on New York State's fracking panel, the one bright light has been the presence of Southwest Energy's Vice President and General Counsel Mark Boling. Boling is bullish on shale gas but his passion for public disclosure and a rigorous and rational regulatory framework, his candor about the perils of certain practices and his honest assessments of the costs and benefits of gas shale extraction have inspired trust and confidence among his fellow panelists. Boling's candor may have made him a pariah in his industry, but the panel's confidence in his integrity is the one thing that might allow us to go forward with recommendations regarding a regulatory scheme that could allow certain kinds of fracking to proceed in New York State. None of us wants to be in the position of getting seduced by sweet and lofty promises that quickly turn into a sour gas and impoverished communities.
Gas fracking flacks routinely make extravagant promises about bringing jobs and income to the depressed rural communities. If those jobs and royalties don't come—the way they have not come for people in Bradford County, PA—New Yorkers will be justifiably angry, as they wonder why the government and our panel did not protect them when there were so many warning signs.
Originally posted in Huffington Post on Oct. 20, 2011.
fracking insights


Gifted individuals are often mishandled and misunderstood by school systems that don't understand them. And most parents have no idea how to help their gifted children achieve their true potential, both socially and vocationally. As a consequence, many gifted people shut down as children and experience creative blocks and other limitations later in life. My own experiences and background in the arts have made me uniquely suited to helping those who are intellectually gifted and those in the creative industries of dance, theater, film, music, art or literature.


Windows PowerShell 2 is the scripting language that enables automation within the Windows operating system. Packed with powerful new features, this latest version is complex, and Windows PowerShell 2 For Dummies is the perfect guide to help system administrators get up to speed.
Written by a Microsoft MVP with direct access to the program managers and developers, this book covers every new feature of Windows PowerShell 2 in a friendly, easy-to-follow format.
Windows PowerShell 2 For Dummies makes this tool easily accessible to administrators of every experience level.
Steve Seguis is a Microsoft Windows systems engineer with more than 12 years of experience managing small- to large-scale Windows environments. He was a Microsoft MVP for Windows Server - Admin Frameworks from 2004–2007, and is a contributing technical editor for Windows IT Pro.


text:
Label: Tiny Global Productions – PICI-0024-LP
Format: Vinyl, LP, Limited Edition
Country: UK & Europe
Released: 08 May 2020
Style: Post-Punk
A1 Thicko Rides Again
A2 The Top Shelf
A3 Neverender
A4 Wicked Winter (Lost In Highland Park)
A5 Then I Felt
A6 The End Began Somewhere
B1 Devil's Due
B2 Everything, Everywhere, All Of The Time
B3 The Other Side
B4 On The Make
B5 Simple Soul
B6 The Desperate Quartet
Recorded At – Faust Studio
Pressed By – GZ Media – 202429E
Alto Saxophone – Kate Armitage (tracks: A1)
Artwork, Design – Fliss Kitson
Bass, Vocals – Andreas Schmid
Double Bass – Mark Bedford (tracks: B6)
Drums, Vocals – Fliss Kitson
Guitar, Vocals – James Smith (38)
Keyboards – Randy Kuntz (tracks: A3, B2)
Mastered By – Stuart Moxham
Producer, Engineer, Mixed By – Andreas Schmid
Violin – Clara Kebabian (tracks: B6)
Vocals – Sanne Richter (tracks: A3)
Vocals, Mouth Organ – Robert Lloyd
Comes in a plain white sleeve. Includes download card.
Nightingales – Four Against Fate (Vinyl) quantity
Categories: 12" Vinyl LP, Distro, Nightingales Tags: 1977, album, Become Not Becoming, Birmingham, Captain Beefheart, CD, double vinyl, film, fliss kitson, Four Against Fate, Frank Skinner, Henry Moore Institute, indie, Iron Man Records, James Nicholls, john peel, King Kong, King Rocker, King Rocker Film, leeds, marc riley, Michael Cumming, music, Nicholas Monroe, Nightingales, No Love Lost, Out Of True, punk, Robert Lloyd, Robert Lloyd and the New Four Seasons, rock, sky, stewart lee, The Clash, the nightingales, the prefects, The Second Best Music Film of 2020, Vinyl, White Riot
An air of the unsettled is a staple of Robert Lloyd's career, from The Prefects's dank dexterity and jittery paranoia of the first Nightingales' release, Idiot Strength, onward through four decades of top-notch recordings. If the unique persona of Lloyd and crew always came across on their ten albums and countless line-ups, it was largely as an acquired taste of the musical cognoscenti. Labels good and bad seemed to feel, at one point or another, a public duty and a point of pride to release a Nightingales album before returning to the business of business.
Four Against Fate is remarkable. It's the work of what's now the band's longest-serving line-up. The instrumental precision of any version of Nightingales has been one of the band's defining hallmarks, but the psychic interplay of a group can take a few albums to kick in with full majesty – here's proof of that. The rhythm section of Fliss and Andi functions now on a purely intuitive level. Jim's work now ranks with that of any guitarist in modern 'rock' music, not just in originality, but also across an egalitarian mass of inspiration. Each member sings. Although Robert's voice functions as the band's superego, Fliss takes lead in several songs. Few bands today sound as much like a single unit as do Nightingales, but this group has the bonus of a distinct and credible musical language, exemplified by The Desperate Quartet, which comes across as both a medieval war march and the anthem of looming apocalypse. When at the song's halfway point, American classical musician Clara Kebabian's violin and Mark Bedford's (of Madness) double bass overtake the Robert, Fliss, Jim and Andi, it's a jawdropper of such intense perversity that it alone defies the listener to not play the album again from the start.
Not that this album lacks 'hits' – The Top Shelf, Everything Everywhere All Of The Time, Devil's Due and The Other Side are stunners. Robert claims Four Against Fate is the first of his album on which he skips no tracks on playback!
Very likely the best album of 2020, and easily the best in a career that's lasted for decades. "Four Against Fate" is the latest in a string of brilliant releases, but here it all comes together like nothing they've done before.
Hot on the heels of an expanded deluxe edition 2xLP reissue of their classic "No Love Lost", "Four Against Fate" was recorded late last autumn at Faust Studio in Scheer Germany by bassist Andi Schmid and mastered by Young Marble Giants' Stuart Moxham. It features Clara Kebabian on violin, Kate Armitage from Hotpants Romance on sax, and Mark Bedford from Madness / Near Jazz Experience on double bass.
Hightlights are too many to list – the immediacy of The Top Shelf, the stirring album closer The Desperate Quartet and the what-the-fuck-is-that madness of "Everything, Everywhere, All Of The Time" . . . any one of them (and several others) could be a hit, and the band will be launching another long UK tour starting in April, two jaunts to Europe and more dates later in the year. 2020 will also see the debut of the Stewart Lee / Michael Cumming film about Robert and the band, "King Rocker", and it's inevitable that there will be some special performances surrounding that.
Mark Eitzel – The Ugly American (Vinyl) Blue 180g USA Edition
Spacemen 3 – Performance (Vinyl)
Spacemen 3 – For All The Fucked-Up Children Of This World We Give You Spacemen 3 (First Ever Recording Session, 1984) (Vinyl)
Nomeansno – Dad / Revenge (Vinyl) 7″
World Party – Private Revolution (Vinyl) £19.50
The Jesus Lizard – Goat (Vinyl) (Deluxe) £23.00
Dag Nasty – Dag With Shawn (Vinyl) £14.00


Which parent has not exulted when baby lifts her hand in an awkward "bye-bye," or shakes her head for the first time to convey an emphatic "no?" All animals have some form of gestural ability in order to communicate within species. My sub-primate golden doodle displays his abilities admirably when he uses a simple communicable action to thumb his nose at a canine neighbor's warning. He lowers his haunches and pees in front of the infuriated barking dog threatened by our proximity to his territory.
During prehistoric times, gesturing and signing were probably the primary means of communication. We can well imagine the Stone Age hunter gesticulating with fingers and eyes for quiet in order to execute a stealthy kill. Among humans, gestures are as natural as talk and they precede human speech in the ontogeny of language. The rolling of eyes in scornful expression is now almost a rite of passage to teenhood.
M.S Thirumalai, editor of Language in India, writes that gestures maintain and enhance "the level of intimacy between individuals." Among other things, he calls it a reflection of the visual attitude of the soul. This observation, though imprecise, does have some credulity, for in extreme and deep moments of connection we often resort to soundless gestural acts like kissing, hugging, smiling or crying. Language somehow becomes inadequate when deep emotions are involved. So while gestures and sign language predate the spoken word, they are never completely eliminated from our vocabulary.
The culture of the Indian subcontinent, in particular, inclines towards gesturing while talking. Hands wave, eyebrows become mobile, and lips shape in slow exaggerated affectation. Dance forms like bharatanatyam rely heavily on the ability to gesture. A reference in Manmohan Ghosh's translation of Natyasastrain 1967 explains the purpose of abhinaya (in dance forms) as illuminating the content of songs or words in a play. So, in a supplemental role, gestures and signing can help advance comprehension.
In a primary role, the benefits to the hearing-challenged are evident, but how about infants and toddlers?
There can be no arguing that babies would be better able to express themselves if they learned a system of communication before their tongues were able to manipulate a coherent language of sounds. Children and infants use signs all the time. Sadly, it is our inability to understand or pay attention that is often the issue. Take for example, the pulling of the ear to indicate an earache, or clenched fists to indicate tiredness, or sticking the tongue in and out of the mouth or smacking the lips to indicate hunger. Experienced parents, in most cases, can gauge what the child needs by the movements that precede the crying session. The benefits of teaching a more formalized system of signing, could possibly take the guesswork out of the process.
Many studies have found that babies use gestures to control, manipulate and understand their environment. "At around nine months of age human infants begin to gesture (and sometimes to vocalize with cries or grunts)," writes socio linguist Elizabeth Bates, "in order to get adults either (i) to do something for them (e.g., get something for them or perform some activity), or (ii) to share attention with them to some object or ongoing event." These types of gesturing, she says, jumpstart the process of communication.
As they grow older, these hand signs and gestures change to a more supplemental role after the acquisition of conventional linguistic skills. Work in neuroscience has shown that the areas in the brain that control the mouth and speech and the areas that control the hands and gestures overlap a great deal and develop together.
Teaching simple gestures and signs to babies before they can talk can springboard developmental milestones, claims Montessori kindergarten teacher Sridevi Natarajan. She watched a friend and her twelve-month-old baby engaging in a signing conversation. Natarajan was intrigued enough to participate in a workshop run by Baby Signs, an organization started by Linda Acredolo of the University of California at Davis, and Susan W. Goodwyn of the California State University at Stanislaus. They found that second graders who had been encouraged to use the Baby Signs signing system during the second year of life had an advantage of 12 IQ points over children who did not use any such system.
Acredolo cautions parents that this is not the reason to sign up for the Baby Signs program. The main reason is to improve the relationship between parent and child and to help babies "share their world with you." In 1996 Acredolo and Goodwyn published Baby Signs: How to Talk to Your Baby Before Your Baby Can Talk, which became a best-selling parenting title, parents being ever ready to clutch at any straw that can perpetuate the dream of superior achievement.
It stands to reason that parents who talk a lot to their children incorporate signs and gestures as part of the communicative package. This increased level of interaction, whether verbal or signing, generally leads to a larger vocabulary. Furthermore, since verbal skills are an essential part of the IQ test, it would naturally result in an increased IQ score.
In a laboratory experiment, it was demonstrated that when two babies (one ten months old and the other nine months) who cried frequently were taught signs, their crying and whining decreased in proportion to their ability to convey commands through signs.
However, the experiment was deemed inconclusive since the experimenters used the strategy of extinction: rewarding only for using the correct signs, never for the crying. In other words the baby was rewarded only for the desired outcome, which biased the results.
With any wildly popular new idea it is important to temper expectations. Esther Wender, a clinical professor of pediatrics at Yeshiva University says that gesturing is "like trying to teach children to walk earlier. It just doesn't make sense neurologically." However, Natarajan passionately believes that signing to infants and toddlers helps their cognitive, verbal and emotional development and "babies throw fewer to no tantrums when they have the ability to express through signs." But wouldn't that also presuppose that verbally agile two- and three-year-olds exist tantrum-free since they have learned an essential vocabulary to express themselves? I only wish that were true!
More information on sign language for babies can be found athttp://babysigns.com.
Jaya Padmanabhan is a prize-winning fiction writer and is currently in the process of writing a novel.


The button placement underneath the bezel is a little awkward and although the keys are clearly labelled, the text isn't backlit. Each menu and sub-menu was in an understandable order and category. Cons Glare due to glossy screen and bezel, awkward button placement, poor viewing angles. There is noticeable colour shift and a loss of detail when viewing the screen from horizontal or vertical angles though the horizontal viewing angles are slightly better. The Desktop is sharp and striking and text has no visible aberrations. The outer edge of the HP m's bezel uses a matte black finish and the controls our housed in the bottom left corner. Although this monitor offers good clarity and colours, excessive glare from the glossy screen and bezel detracts from the overall user experience.
Each menu and sub-menu was in an understandable order and category. The 3ms response time means it isn't as quick as some monitors most new LCD monitors have a 2ms response time but the HP m handles fast motion superbly, with no ghosting visible.
Combined with the native Full HD resolution of x, the HP m is ideal for use with the latest high-definition gaming consoles, such as Microsoft's Xbox and Sony's PlayStation 3. If you can afford the price tag, it is well worth the money. It out performs any other laptop I have tried for gaming, and the transportable design and incredible display also make it ideal for work.
There is noticeable colour shift and a loss of detail when viewing the screen from horizontal or vertical angles though the horizontal viewing angles are slightly better.
It has a rounded, glossy black bezel with silver edging and a curved silver base. Cons Glare due to glossy screen and bezel, awkward button placement, poor viewing angles.
The button placement underneath the bezel is a little awkward and although the keys are clearly labelled, the text isn't backlit. HP claims the design of the base allows you to maximise desk space. Sign in with LinkedIn Sign in with Facebook. Would you buy 2309, Although this monitor offers good clarity and colours, excessive glare from the glossy screen and bezel detracts from 239m overall user experience.
Sign up to gain exclusive access to email subscriptions, event invitations, competitions, giveaways, and much more. Touch screen visibility and operation was great and easy to navigate. HP claims the use of BrightView technology reduces glare, but even after adjusting the myriad of settings available it up a huge problem in the office environment where we tested.
The built-in speakers aren't the worst we've seen in a monitor, but any self-respecting 2039m will use external speakers. We connected the LCD monitor to both an Xbox and a Blu-ray player, and the results were impressive. Image quality when using the DVI input is also impressive. Membership is free, and your security and privacy remain protected.
Colour reproduction is excellent: Unfortunately, the bezel and the glossy surface of the display make reflections a problem.


The Crafar family is taking a new tack in its fight to stay on the farms it has been ordered to leave.
The receivers of the Crafar farming operation have taken legal action to evict Allan Crafar and other family members from three houses on two of the 16 farms that have been put up for sale.
Mr Crafar has now told a judge at the High Court in Rotorua that the family has tenancy agreements with its companies, which are in receivership, to stay on the land for a peppercorn rental.
One of the receivers, Michael Stiassny, says they had not heard of the agreements until last week.
He says the court has given Mr Crafar until August the sixth to produce the tenancy agreements.
The documents will then be examined by police forensic experts.


Thank you for visiting our website. Stephen P. O'Hare PLLC is an experienced attorney in Poughkeepsie, NY. We help clients with a variety of legal issues, including personal injury, accidents, estate planning, wills and trusts, criminal matters, construction law and commercial, industrial, and residential real estate transactions.


Remember yesterday we told you that the ETAPA service technician said he'd be out in the morning to let us know what is wrong with our connection? Ok, he did show up and the problem was with the phone line that connects to the modem box. The tech gave us a new line and he left, problem solved…so we thought.
We thought, "Great, the Internet is back, now we can get back to work". I began uploading a new video to You Tube when in the middle of the upload the Internet went back out again, so it is not the Internet line; it has to be something else. The problem was solved for about an hour.
We called ETAPA again and the lady on the phone said someone would be out Friday to see what the problem is. Friday! But this is only Tuesday; what are we supposed to do until then? After four days of not being able to get on the Internet it tends to get a little bit frustrating and patience runs a bit thin.
We are asked all the time "how is the Internet in Ecuador", well, there are problems with it at certain times. If you make a living online, we suggest move somewhere else; Our experience dictates we are not able to recommend Ecuador; the Internet is not reliable enough for us to encourage such a move for anyone that has to have good internet all the time.
Over the last thirteen months we have had major issues with the connection on four different occasions, where we don't have Internet for days, sometimes weeks at a time. Today is the fourth day with no Internet for us. Sometimes, mostly in the mornings, we can stream videos, but by afternoon and evening, forget about it. It isn't going to happen with ETAPA. We have heard mixed reviews about CABLETV Internet of Ecuador. Some have said it is better than ETAPA, some have said it is terrible Internet.
At the moment we happen to be using a CLARO wireless rechargeable stick, which we use for emergencies such as this. Down time is not going to get us down. We're back up on the Internet even though our primary Internet happens to be out of operation. A guy from ETAPA supposed to be coming out again tomorrow…we'll see.
We're not giving up hope. After all this, our idea is to order a second internet company. That is, have two internet companies providing service to our house. First we'll have to see if they will install a second internet in the same house. We will keep you posted on how that turns out for us.


Embracing Balance: Can this finally be Summer?
Can this finally be Summer?
Day Two of glorious sunshine! Yaaaieeeeee. Loves it!
When my alarm went off this morning I had no trouble getting up because I could tell it was a beautiful day (might have been the sun blaring into my eyes through the crack in the curtains…). Got into my shorts, grabbed my coffee, and headed straight for the front steps with my magazine. Perfect way to start the day.
After getting my morning work out of the way, I was back out to the steps with my second cup of coffee and my breakfast. A delicious cheese scone with sliced tomatoes.
When I got back I pounded out some more work, then made lunch. Rummaging through the fridge I found the makings for an awesome hot weather meal…salmon pasta salad.
I mixed the leftover Smart macaroni with the extra salmon filet from last night, some red onion, some carrots, a dollop of light mayo, a squirt of grainy dijon, and a sprinkle of dill, sea salt, and pepper.
At the last minute I decided to stir in some baby spinach and a squirt of lemon. Made enough for two servings…yum yum.
Onto the back deck I went. With a POM spritzer and some Kashi crackers for the crunch factor.
A couple more hours of work, then back out to lay on the deck. This time I laid out a blanket and some pillows so I could get the sun on my back. Pretending I was at the beach I decided I needed a snack so broke into this little bag of Baked Old Dutch chips…deeeelish.
I've also snacked my way through about 3/4 of this 1 pound box of local strawberries today. They are just so good I can't help but eat one (or 5!) every time I walk by.
After finishing work for the day, I went for a 20 minute walk around the neighborhood. I will be so glad to go back to Booty Camp tomorrow night…this lack of real exercise since Wednesday is driving me batty. Cross your fingers for my back!
Dinner had to be something light and simple. Salad was on the menu for sure but I needed to cook up a chicken breast too. There was no way I was turning the stove on, so back to the BBQ I went.
Huge salad: bagged romaine, tomatoes, cukes, red onion, yellow pepper, leftover grilled beets, Ranch dressing, a squirt of BBQ, grated organic old white cheddar.
Mixed all together, the flavors were perfect for a hot Summer day.
Now that it's cooled down, I puttered in my little garden for a bit, cleaned the upstairs bathroom (including tub!), dusted upstairs, and vacuumed upstairs. Whew. I rewarded myself with ice cream I bought at Planet Organic a couple of weeks ago. Chocolate Peanut Buttery goodness…but had I noticed the nutritional information I probably wouldn't have bought it…that calorie content is like double what normal run of the mill regular ice cream would be!
Don't forget I'm looking for guest posts. Come out, come out wherever you are! I want to hear from you!!
It was so gorgeous out yesturday, figures its gray out this am. Your eats were perfectly Summer, yummy!
July 21, 2009 at 4:36 a.m.
Yup yesterday was awesome! I wanted nothing more than to watch the ships sail out, but I had to come home and rest the ol' hip. It was so nice out!
July 21, 2009 at 4:59 p.m.


text:
The Taking of Channel 9
Women in Oaxaca Make a Stand for Local Teachers
It all started with a teachers strike.
In the beginning, the civil disobedience was not organized primarily by women. Oaxaca teachers decided to strike to demand higher federal and state education budgets. They set up a protest camp in Oaxaca City, a tent city that filled the touristy town square and stretched out for blocks, housing tens of thousands of teachers from across the state.
Oaxaca Governor Ulises Ruiz Oritz refused to meet with the teachers' union or answer their demands. Instead, he sent state riot police using tear gas and helicopters to dismantle the striking teachers' camp, leaving scores of men, women, and children injured.
The city exploded. Thousands took to the streets to help the teachers, tend the injured, and offer food and water. But to everyone's surprise, these citizens went one step further--they counterattacked, retook the town square, and drove the police out of town.
This spontaneous rejection of police violence, along with the outpouring of support for the teachers, ignited a five-month civil disobedience uprising. It would put a half million people on the streets in marches and tens of thousands in protest camps across Oaxaca City, paralyze the state government, and send the governor into hiding.
Indigenous organizations, human rights groups and local unions joined the teachers in calling for the Governor's resignation for ordering the police raid. Initially, protest leaders were was almost entirely male, with women relegated to lesser roles. Undaunted, women formed neighborhood groups in order to participate and joined marathon discussions that guided the protesters' actions. In the summer of 2006, they cooked up the idea of a women-only march.
The march drew some 5,000 women, all banging on pots and pans with meat tenderizers, ladles, and soup spoons. The raucous cacophony had the women so jazzed that when they reached their destination (the protester-occupied town square), they decided to keep going, to the state-owned television station, Channel 9. The only statewide local station, Channel 9 failed to report on the police violence and later presented the protesters as vandals and hooligans. At first the women demanded only an hour on television to tell their version of the events and why they wanted Ruiz out of office. But the station director refused. The women asked for less time, then even less, but were repeatedly rebuffed. Finally, they walked past the director, with pots and pans in hand, and took over the station.
Employees had taken the station off the air as the women stormed the office. Now the women scrambled to get the station back on the air, trying to figure out the cameras and sound system, before the police came to retake the station.
But the police did not come. Instead, thousands of residents from the surrounding neighborhood flooded the streets to guard the station, taking over city buses and parking them across the street to block all approaching traffic.
Within three hours, for the first time in Mexican history, a protest movement occupied a state television station and broadcast live. Viewers saw a tight group of women without makeup or designer dresses, pots and pans still in hand, all facing the camera. Their message: if the media insist on airbrushing state violence from the news and distorting social protest into an "urban guerrilla" movement, then the people will take the media in order to tell their own story of suffering, police repression, and organizing social protests.


It was the morning of my birthday and I found myself craving blueberry muffins. I don't remember if maybe it's some sort of tradition, but it was definitly something that I just had to have. Problem is, I haven't quite mastered a gluten-free, vegan moist muffin recipe yet. You can call it birthday luck, or a birthday wish come true because these Blueberry Muffins came out so darn good!
The recipe is so simple, which in my house is a must. So what's the secret? What makes these muffins so moist and delicious? Yogurt! Dairy-free blueberry yogurt! I also like to add crystal sugar on the top for that added sweet crunch.
From making other vegan bloggers bakery, I've learned a method to make baked goods better… creating a sort of "buttermilk" from adding apple cider vinegar to your almond milk. It makes the milk curdle ever so slightly. I'm not entirely sure why this makes a difference, but it truly makes my gluten-free, vegan bakery moister and just a little less dense.
Let's take a peek at what's in these delicious gluten-free, vegan Blueberry Muffins and then check out the full recipe so you can enjoy these Blueberry Muffins as well!
These look so bright and refreshing!!! I love your shots and styling 🙂 I'm glad I found you!


Arriving at the venue to find the performers chatting cheerfully to the audience at the bar, you may feel like you've walked in on a rehearsal. It's an impression that stays with you as the company exhaustively explores a single suggestion from the audience with ambling sketches and flippant philosophy. The results are uneven. There's plenty of surreal imagination at work, but despite a gloriously silly game of Chinese whispers in five languages, there's not enough inspiration to sustain the show. The inexperienced players spend too long groping for new angles, letting scenes fade away without a punchline. Though spontaneous, this feels less like a structured, theatrical game and more like a group of friends playing.
Kiwi Bar at Walkabout, 14 – 20 Aug, 8.45 pm (9.45 pm), free, fpp259.


35 Drumsheugh Gardens, Edinburgh, Midlothian. EH3 7RN.
With a history of over 150 years, the Bonham is set within a lovely triangle of traditional town houses. Relatively inconspicuous from the outside, the hotel has a very great deal to offer on the inside.
All of the bedrooms are equally stylish and extremely comfortable, and some of them offer spectacular views over the Dean Village and Edinburgh's historic New Town centre. Comfy beds with crisp white sheets and fluffy pillows guarantee a good night's sleep after a busy day. For those having to catch up on some work, there are desks with plenty of power sockets. The bathrooms are well equipped with baths and showers, modern appliances, luxury toiletries, and soft, warm towels.
Dining at the Bonham is a wonderful experience. The stylish dining room provides the perfect setting for a memorable evening. Choosing between the A la Carte Menu and the Market Menu can be a hard decision as as all of the dishes look and taste wonderful. The head chef Maciej Szymik and his team definitely do their best to keep the choices interesting. By using Scottish ingredients and traditional dishes, and giving them a lift with international influences, such as Asian cuisine, the elegantly designed plates are an experience not to be missed. The afternoon teas are also to be commended.
The bar is stocked with Scotch Whiskies, with expert advice to ensure that you choose your personal best.
Located in Edinburgh's West End of Edinburgh, both the City Centre and Princess Street are only a five minute walk away and therefore shopping, museums, theatres and art galleries are all within walking distance. Moreover, the Bonham's staff are always more than happy to help you arrange whatever it is that you are looking for.


Pork Carnitas Slow Cooker - "Authentic Shredded Mexican Meat" - Easy!
Today I want to share a shredded meat recipe from Mexico, slow-cooked pork carnitas. When we were traveling through Mexico tender shredded pork carnitas was one of my favorites to buy from the street markets pulled fresh from the cooker.
Typically there would be a large pot of pork carnitas meat next to a grill. When a customer ordered a pork taco, the cook would scoop the most delicious shredded pulled pork meat from the pot and place it on a hot grill.
Cooking on the grill would create delicious crispy bits of pork and give the meat a slightly smoky flavor.
When the shredded pork was sufficiently browned it was placed between two soft tortilla's. We would then be able to add our own sauces from the salsa bar.
Even the most humble street vendor had an impressive range of sauces. We found eating from these stalls a real learning experience.
By trial and error, we discovered that the tomatillo salsa verde was usually slightly milder than the red sauces and perfect for our taste buds over our pulled pork carnitas.
Even with our friend's local knowledge we never quite knew how hot each of the homemade salsa's would be.
I'm sure the locals loved seeing tourists add too much hot sauce to their pork carnitas, and then turn red in the face, whilst gasping for breath.
If there is one thing I enjoy more than eating local food when I'm on holidays it's learning to cook local food.
The recipe for slow cooker pork carnitas (or Mexican shredded pork) I'm sharing today is one that I brought back from Mexico.
It's really simple and the use of a slow cooker means that it requires no input or attention during the cooking time.
The ingredients for your pork carnitas recipe is readily available from most stores no matter where you are in the world.
To make the process even easier prep the pork carnitas the night before and refrigerate.
Next morning all that's left for you to do is put the dish into the slow cooker and turn it on before you leave for work.
When you get home you will be greeted with the most amazing food aroma's form your delicious Mexican pork carnitas.
Slow cooked Pork Carnitas is an authentic "Mexican Pulled Pork" recipe. They taste exactly the same as a market stall in Mexico!
Shred the pork with two forks. Put the meat side whilst you make the sauce.
Add enough pulled pork to cover the base of the pan.
I usually serve the pork carnitas with homemade soft corn tortillas. If you're in a rush, you could always use store bought tortilla's. I find tortilla making very therapeutic.
While you're at the shops buy a "dry slaw" add lime juice and fresh coriander (cilantro) for a fresh salad to accompany the pork carnitas.
Then if you want the meal to be extra special, whip up my 5 minutes Mexican Guacamole.
The combination of flavors such as lime, coriander, and citrus gives a very fresh Mexican feel to the shredded pork. It could easily be served in Summer or Winter.
Until I can return to Mexico, I will have to be satisfied with making these pork carnitas myself. I will continue to dream about watching the carnitas meat cooking at the street market. I loved smelling that rich meaty aroma, and trying to make a wise salsa choice!
Make sure you pin this Slow Cooker Pork Carnitas recipe for when you next crave a pulled or shredded Mexican meat feast.


After Blue Shield rate increase disaster, now the "child only health insurance policies" are another disaster for California residents.
In the last quarter of 2010 with the exception of Blue Shield no insurance carrier offered anymore child only policies for children under 19 years, due to the fact that accordingly to the healthcare reform they had to accept all children, even the unfortunate one's with severe health problems. Than the insurance commissioner threatened, if the carriers would not offer child only policies, the carriers would be banned for 5 years in the CA market.
Now most carriers have an open enrollment window for "child-only policies" for children under 19 years, which most carriers offer from January 1st to February 28th of 2011.
After investigating the individual insurance plans, I can only find PPOs plan with very high deducible Anthem, Aetna offers a $3500 deductible Plan. Blue Shield , Cigna offers a $ 5000 deductible -Healthnet and Anthem are offering also a HMO but rates are higher than any available PPO plans.
Before the Healthcare reform we were able to offer insurance PPO plan with $ 500 and $ 1000 deductible – these plans were very affordable for children. They are not existing anymore and this creates a hardship for all families in CA.
I do understand the insurance carrier having to protect themselves from the financial loss of huge claims for children with health problems, but it is not fair that all CA children cannot find suitable policies.
I will not offer a child policy with a deductible of $ 3500 or $ 5000, it is to easy for children to get sick and have to go to the emergency. Just to open the door of an emergency room cost $ 500, Parents should not take this risk!
Please offer more insurance plans for children under the age of 19 years.
The CA State should support the insurance companies with the unhealthy children in CA, and not just demand that the insurance companies carry the burden of the unhealthy children.
Our children are our future and we must make sure that they are protected!


Eng. Osama Mahdy serves as Chairman of the Board & Managing Director, representing Petrochemical Holding Company since January 25, 2016. Prior to that, he served as Marketing Research & Pricing General Manager.
Mrs. Inas Al Syed Mohamed Al Sayed serves as Non-Executive Member of the Board of Sidi Kerir Petrochemicals Company SAE, representing National Investment Bank since February 23, 2016.
Mr. Jabir Ahmed Hasan serves as Non-Executive Member of the Board of Sidi Kerir Petrochemicals Company SAE, representing Egyptian Petrochemicals Company since September 20, 2016.
Mr. Mohammed Mustafa Hasan serves as Non-Executive Member of the Board of Sidi Kerir Petrochemicals Company SAE, representing Employees Social Insurance Fund for Private & General Sector since September 28, 2016.
Mr. Sami Abdul Hadi Mohammed serves as Non-Executive Member of the Board of Sidi Kerir Petrochemicals Company SAE, representing the Employee Social Insurance Fund since January 15, 2016.
Mr. Hasni Husain Hamd Musharaf serves as Non-Executive Member of the Board of Sidi Kerir Petrochemicals Company SAE, representing Misr Insurance.
Eng. Mohammed Hasan Shouqi Saafan serves as Non-Executive Member of the Board of Sidi Kerir Petrochemicals Company SAE, representing Egyptian Petrochemicals Holding Company since October 17, 2016.