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Cellular/Molecular
Nongenomic Glucocorticoid Inhibition via Endocannabinoid Release in the Hypothalamus: A Fast Feedback Mechanism
Shi Di, Renato Malcher-Lopes, Katalin Cs. Halmos and Jeffrey G. Tasker
Journal of Neuroscience 15 June 2003, 23 (12) 4850-4857; DOI: https://doi.org/10.1523/JNEUROSCI.23-12-04850.2003
Shi Di
1Division of Neurobiology, Department of Cell and Molecular Biology, and 2Neuroscience Program, Tulane University, New Orleans, Louisiana 70118-5698
Renato Malcher-Lopes
Katalin Cs. Halmos
Jeffrey G. Tasker
Glucocorticoid negative feedback in the brain controls stress, feeding, and neural-immune interactions by regulating the hypothalamic—pituitary—adrenal axis, but the mechanisms of inhibition of hypothalamic neurosecretory cells have never been elucidated. Using whole-cell patch-clamp recordings in an acute hypothalamic slice preparation, we demonstrate a rapid suppression of excitatory glutamatergic synaptic inputs to parvocellular neurosecretory neurons of the hypothalamic paraventricular nucleus (PVN) by the glucocorticoids dexamethasone and corticosterone. The effect was maintained with dexamethasone conjugated to bovine serum albumin and was not seen with direct intracellular glucocorticoid perfusion via the patch pipette, suggesting actions at a membrane receptor. The presynaptic inhibition of glutamate release by glucocorticoids was blocked by postsynaptic inhibition of G-protein activity with intracellular GDP-β-S application, implicating a postsynaptic G-protein-coupled receptor and the release of a retrograde messenger. The glucocorticoid effect was not blocked by the nitric oxide synthesis antagonist NG-nitro-l-arginine methyl ester hydrochloride or by hemoglobin but was blocked completely by the CB1 cannabinoid receptor antagonists AM251 [N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide] and AM281 [1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-4-morpholinyl-1H-pyrazole-3-carboxamide] and mimicked and occluded by the cannabinoid receptor agonist WIN55,212-2 [(β)-(+)-[2,3-dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-naphthalenylmethanone mesylate], indicating that it was mediated by retrograde endocannabinoid release. Several peptidergic subtypes of parvocellular neuron, identified by single-cell reverse transcripton-PCR analysis, were subject to rapid inhibitory glucocorticoid regulation, including corticotropin-releasing hormone-, thyrotropin-releasing hormone-, vasopressin-, and oxytocin-expressing neurons. Therefore, our findings reveal a mechanism of rapid glucocorticoid feedback inhibition of hypothalamic hormone secretion via endocannabinoid release in the PVN and provide a link between the actions of glucocorticoids and cannabinoids in the hypothalamus that regulate stress and energy homeostasis.
retrograde messenger
parvocellular neuron
membrane receptor
corticotropin-releasing hormone
thyrotropin-releasing hormone
vasopressin
oxytocin
Glucocorticoids secreted by the adrenal cortex exert inhibitory effects on several hypothalamic neuroendocrine systems, including but not limited to the negative feedback regulation of the hypothalamic—pituitary—adrenal (HPA) axis by suppression of the secretion of corticotropin-releasing hormone (CRH) from neurons of the hypothalamic paraventricular nucleus (PVN) (Herman et al., 1996; de Kloet, 2000). The glucocorticoid negative feedback regulation of the HPA axis occurs acutely by a relatively rapid inhibition of CRH release and chronically by down-regulating CRH and vasopressin (VP) expression in PVN neurons (Keller-Wood and Dallman, 1984). Classical glucocorticoid actions, like those of other steroid hormones, are mediated by binding to intracellular receptors and regulation of gene transcription (Falkenstein et al., 2000). Recent evidence in different species suggests that rapid glucocorticoid actions are mediated by membrane receptors and activation of nongenomic signaling mechanisms. Thus, glucocorticoids have been shown to bind specifically to cellular membrane sites (Suyemitsu and Terayama, 1975; Harrison et al., 1979; Orchinik et al., 1991) and to induce rapid influences on electrolyte movement across cellular membranes (Avanzino et al., 1987; Hua and Chen, 1989; ffrench-Mullen, 1995). Although the molecular identity of the membrane-associated receptors for corticosteroids is still unresolved, a high-affinity binding site for corticosterone, which appears to meet all of the criteria for a functional membrane-associated corticosteroid receptor, has been partially purified and characterized in neuronal membranes from amphibian brain (Evans et al., 2000). However, it remains controversial where in the mammalian brain and through which mechanisms glucocorticoids mediate their rapid feedback regulation of the HPA axis. Here, we show that glucocorticoids exert a rapid inhibitory effect on glutamate release onto identified parvocellular neurons of the PVN, including CRH neurons, by activating a G-protein-dependent signaling mechanism that leads to the retrograde release of an endocannabinoid and results in the suppression of synaptic glutamate release.
Slice preparation. Male Sprague Dawley rats (3–5 weeks of age; Charles River Laboratories, Wilmington, MA) were used in these experiments according to a protocol approved by the Tulane University Institutional Animal Care and Use Committee and in accordance with United States Public Health Service guidelines. Rats were deeply anesthetized with pentobarbital sodium (50 mg/kg body weight) and decapitated. The brain was quickly removed from the cranial cavity after cutting the optic nerves and immersed in cooled (1–2°C), oxygenated artificial CSF (aCSF). The composition of the aCSF was as follows (in mm): 140 NaCl, 3 KCl, 1.3 MgSO4, 1.4 NaH2PO4, 2.4 CaCl2, 11 glucose, and 5 HEPES, pH adjusted to 7.2–7.3 with NaOH. The hypothalamus was blocked and the caudal surface of the block was glued to the chuck of a vibrating microtome (World Precision Instruments, Sarasota, FL). Two coronal hypothalamic slices (350 μm) containing the PVN were sectioned, bisected along the midline (i.e., at the third ventricle), and submerged in a holding chamber in oxygenated aCSF at room temperature, where they were allowed to equilibrate for >1.5 hr before being transferred to the recording chamber.
Electrophysiological methods. Patch pipettes were pulled from borosilicate glass (1.65 mm outer diameter, 1.2 mm inner diameter; KG33; Garner Glass, Claremont, CA) with a Flaming/Brown P-97 micropipette puller (Sutter Instruments, Novato, CA) to a resistance of 3–4 MΩ. The pipette solution contained (in mm): 120 K-gluconate, 10 KCl, 1 NaCl, 1 MgCl2, 1 EGTA, 2 Mg-ATP, 0.3 Na-GTP, and 10 HEPES, pH adjusted to 7.3 with KOH. The osmolarity of the solution was adjusted to 300 mOsm with 20 mm d-sorbitol.
Hemislices were transferred one at a time from the holding chamber to a submersion recording chamber, secured to the floor of the chamber with AgCl2-coated wire, and allowed to equilibrate for at least 15 min before recordings. Medial PVN neurons were visualized with a cooled CCD camera using infrared illumination and differential interference contrast optics and patch-clamped under visual control. After achieving the whole-cell configuration, series resistance and whole-cell capacitance were adjusted and continuously monitored during experiments. Recordings with unstable series resistance were discarded. Whole-cell recordings were performed in voltage-clamp mode using an Axopatch 1-D amplifier (Axon Instruments, Foster City, CA) and monitored continuously on a digital storage oscilloscope (Hitachi, Tokyo, Japan). All data were low-pass-filtered at 2 kHz, converted to digital video format at 22 kHz using a Neuro-Corder (NeuroData Instruments, New York, NY), and stored on videotape for off-line analysis. Selected data were subsequently digitized off-line at 4 kHz and recorded on a personal computer using the Digidata 1200 interface and pClamp 7.0 software (Axon Instruments). Miniature EPSCs (mEPSCs) in putative parvocellular PVN neurons were recorded in the presence of the voltage-gated sodium channel blocker tetrodotoxin (TTX) (1 μm) at a holding potential of -60 mV at 22–24°C. To focus on the fast, nongenomic actions of glucocorticoids, 3 min epochs of mEPSCs were collected just before bath application of glucocorticoids and after 7 min of glucocorticoid application. The frequency, amplitude, and decay time (defined as the time from peak to the point at which the mEPSC had decayed by 63%) of mEPSCs were analyzed using the Minianalysis 5.0 program (Synaptosoft, Decatur, GA). Statistical analyses were performed using the Student's paired t test for within-cell comparisons, the Student's unpaired t test for between-group comparisons, and the Kolmogorov—Smirnov nonparametric test for within-cell comparisons of mEPSC distribution; p values of <0.05 were considered significant.
Type II, putative parvocellular PVN neurons were differentiated from type I, putative magnocellular PVN neurons on the basis of their location in the PVN, morphology, and the absence of transient outward rectification (Hoffman et al., 1991; Tasker and Dudek, 1991; Luther et al., 2000). The presence of transient outward rectification was assessed with a current-clamp protocol consisting of a series of progressively more depolarizing current injections from a hyperpolarization to near -100 mV. Transient outward rectification, found only in type I putative magnocellular neurons, was identified as a pronounced dampening of the membrane-charging curve that was associated with a delay in the onset of depolarization-induced action potentials. Type II neurons were divided into putative neurosecretory and non-neurosecretory parvocellular neurons on the basis of the expression or lack of expression of a calcium-dependent low-threshold spike (Luther et al., 2002). Only putative parvocellular neurosecretory neurons, which lacked a low-threshold spike, were included in this study.
Drug application. Water-soluble forms of the steroids dexamethasone (DEX) (0.01–100 μm), corticosterone (1 μm), and cholesterol (5 μm) (Sigma-Aldrich, St. Louis, MO) were directly dissolved in aCSF to final concentrations and applied in the bath perfusion. The DEX—bovine serum albumin (BSA) conjugate (10 μm) (purified to remove free steroids by the supplier; Steraloids, Newport, RI) was dissolved in aCSF with 25% β-cyclodextrin (Sigma-Aldrich) as a carrier to increase its solubility. The concentration of DEX—BSA, (10 μm) was selected to obtain an effective concentration of dexamethasone of ∼1 μm, because the BSA conjugation reduces the effectiveness of receptor binding by a factor of ∼10. TTX (1 μm) (Sigma-Aldrich), the nitric oxide (NO) synthase (NOS) inhibitor NG-nitro-L-arginine methyl ester hydrochloride (L-NAME) (50 μm) (Tocris Cookson, Ellisville, MO), and the carbon monoxide (CO)—NO scavenger hemoglobin (20 μm; Sigma-Aldrich) were dissolved in sterile water and stored in 10 mm stock solutions at -20°C. The physiologically inactive steroid isopregnanalone (5 μm), the intracellular type I and type II corticosteroid receptor antagonists spironolactone (10 μm) and mifepristone (RU486) (10 μm), the protein kinase blockers staurosporine (1 μm) and bisindolylmaleimide (GF109203X) (0.5 μm) (Sigma-Aldrich), the cannabinoid receptor agonist (R)-(+)-[2,3-dihydro-5-methyl-3-(4-morpholinylmethy)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-naphthalenylmethanone mesylate (WIN55,212-2) (1 μm), and the cannabinoid receptor antagonists N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (AM251) (1 μm), 1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-4-morpholinyl-1H-pyrazole-3-carboxamide (AM281) (10 μm), and 6-iodo-2-methyl-1-[2-(4-morpholinyl)ethyl]-1H-indol-3-yl] (AM630) (1–10 μm) (Tocris Cookson) were stored as 10 mm stock solutions in DMSO at -20°C and dissolved to their final concentrations in aCSF before bath application. The cannabinoid analogs were also dissolved with 25% β-cyclodextrin. The DMSO and β-cyclodextrin solutions without the cannabinoids or glucocorticoids had no effect on mEPSCs at the concentrations used. The nonhydrolyzable guanylyl nucleotide GDP-β-S (500 μm) (Sigma-Aldrich) and, in some cases, dexamethasone (1 μm) were included in the patch solution for intracellular application. To prevent drugs destined for intracellular application from leaking into the extracellular space before a membrane seal was obtained, the tips of the patch pipettes were filled first with regular patch solution and then back-filled with the drug solution.
Single-cell reverse transcription-PCR. The single-cell reverse transcription (RT)-PCR procedure was similar to that described by Surmeier et al. (1996), with minor modifications. The cell contents were aspirated into the patch pipette under visual control, taking care to avoid aspiration of the nucleus, and transferred into 5 μl of a lysis buffer containing (in μl) 2.9 of diethyl pyrocarbonate (DEPC)-treated water, 0.7 of BSA, 0.7 of oligo-dT (0.5 μg/μl), and 0.7 of RNasin (40 U/μl), which was maintained on ice before and after harvesting of the cytoplasm. The cells were stored at -80°C or used immediately for RT. The lysate-buffer mixture was heated to 70°C for 10 min and incubated on ice for 1 min, after which 15 μl of final RT master mix, containing (in μl) 8.0 of DEPC-treated water, 2 of 10× first-strand buffer, 2 of MgCl2 (25 mm), 2 of DTT (0.1 m), and 1 of mixed deoxy NTPs (dNTPs) (10 mm), was added. Single-stranded cDNA was reverse transcribed from the cellular mRNA by adding 0.7 μl of SuperScript II reverse transcriptase (200 U/μl) and incubating at 42°C for 50 min, after which the reaction was terminated by heating to 70°C for 15 min. The RNA strand in the RNA—DNA hybrid was removed by adding 0.5 μl of RNase H (2 U/μl) for 20 min at 37°C. All reagents were obtained from Invitrogen (Madison, WI).
PCR amplification was performed with a thermal cycler (MJ Research, Watertown, MA) using a fraction of the single-cell cDNA as a template. Reaction mixtures contained (30 μl final volume): 2.2 mm MgCl2, a 0.5 mm concentration of each of the dNTPs, a 1.5 μm concentration of primers, 2.5 U of Taq DNA polymerase, 2.9 μl of 10× buffer, and 4 μl of the cDNA template made from the single-cell RT reaction. The thermal cycling program was set at 94°C for 45 sec, 60°C for 45 sec, and 72°C for 1.10 min for 50 cycles.
Primers for CRH and thyrotropin-releasing hormone (TRH) were designed in our laboratory using Primer Select 4 (DNAStar, Madison, WI), and primer designs for VP, oxytocin (OT) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were taken from Glasgow et al. (1999). All primers were synthesized by Integrated DNA Technologies (Coralville, IA). The following primers were used: CRH, 5′-GCCCCGC AGCCGTTGAA-3′ and 5′-GACCGCCTCCCTCTCTCCAG-3′ (326 bp product); TRH, 5′-GGGGACCTCGGTGCTGCCTTAGAC-3′ and 5′-CTGCCGCTTGACTTGGGGGACATC-3′ (276 bp); OT, 5′-GACGGT GGATCTCGGACTGAA-3′ and 5′-CGCCCCTAAAGGTATCATC ACAAA-3′ (462 bp); VP, 5′-CCTCACCTCTGCCTGCTACTT-3′ and 5′-GGGGGCGATGGCTCAGTAGAC-3′ (440 bp); GAPDH, 5′-GGAC ATTGTTGCCATCAACGAC-3′ and 5′-ATGAGCCCTTCCACGAT GCCAAAG-3′ (441 bp). Each of the primers was used for every cell analyzed. Final PCR products were visualized by staining with ethidium bromide after separation by electrophoresis in 1.5–2% agarose gels. The forward and reverse GAPDH primers were designed to span an intron to detect genomic DNA in each cellular template. In addition to the single-cell GAPDH RT-PCR, whole hypothalamic slices were run in parallel with the single-cell reactions as positive controls for the primers. Negative controls for contamination from extraneous and genomic DNA were also run for every batch of neurons by running pure water or by omitting the RT step from the reaction, respectively. None of the cells tested showed a genomic DNA PCR product.
To characterize the fast effects of glucocorticoids on putative neurosecretory parvocellular PVN neurons, we performed whole-cell patch-clamp recordings in neurons located in the medial parvocellular region of the PVN in acutely prepared hypothalamic slices. A total of 131 putative neurosecretory parvocellular PVN neurons, identified on the basis of their visualized position within the PVN and on electrophysiological criteria described in Materials and Methods, were recorded in acute hypothalamic slices from 80 rats. mEPSCs were recorded at a holding potential of -60 mV, which was close to the calculated chloride equilibrium potential (ECl = -63.3 mV), in the presence of TTX (1 μm). They were blocked by the glutamate receptor antagonists AP-5 (50 μm) and DNQX (30 μm) (n = 4) but were not affected by the GABAA receptor antagonist bicuculline methiodide (30 μm) (n = 6), indicating that they were mediated by synaptically released glutamate. Data were collected after an initial 10 min baseline period was established during which the amplitude and frequency of mEPSCs were stable. The mEPSCs recorded under control conditions ranged in amplitude from 8.2 to 36.9 pA, in frequency from 0.34 to 6.95 Hz, and in decay time from 1.8 to 4.5 msec.
Glucocorticoids inhibit glutamate release
The glucocorticoids dexamethasone and corticosterone applied in the bath perfusion had no effect on membrane-holding current or input conductance in any of the neurons tested but suppressed glutamatergic synaptic activity within 3–5 min in 22 of 25 putative parvocellular neurosecretory neurons of the PVN (i.e., >10% change in mEPSC frequency). Dexamethasone (1 μm), a selective synthetic glucocorticoid receptor agonist, caused a significant decrease in the frequency (3.11 ± 0.52 to 1.83 ± 0.39 Hz; 58.8 ± 4.8% of baseline; p < 0.01; n = 18), but not the amplitude (24.77 ± 1.95 vs 24.10 ± 1.76 pA; 98.18 ± 1.5% of baseline; p = 0.13) or decay time (3.52 ± 0.17 vs 3.68 ± 0.16 msec; 105.0 ± 4.5% of baseline; p = 0.32) of mEPSCs (Fig. 1a—c). The inhibitory effect of dexamethasone on mEPSC frequency had a rapid onset (3–5 min) and was dose dependent, with a threshold concentration between 10 and 100 nm and a saturating concentration between 1 and 10 μm (Fig. 1d). Corticosterone (1 μm) had a similar inhibitory effect on mEPSCs, causing a decrease in the mEPSC frequency (1.90 ± 0.57 to 1.39 ± 0.43 Hz; 72.4 ± 4.6% of baseline; p < 0.01; n = 7) (Fig. 1e), without affecting either the mEPSC amplitude (33.16 ± 0.14 vs 31.75 ± 0.57 pA; 97.0 ± 2.4% of baseline; p = 0.13) or decay time (4.06 ± 0.85 vs 3.79 ± 0.76 msec; 98.0 ± 6.13% of baseline; p = 0.36). The effect of glucocorticoid did not reverse after 60 min of washout of the steroid. The effect was steroid-specific, because the corticosteroid precursor cholesterol (5 μm) and the physiologically inactive steroid isopregnanolone (5 μm) had no effect on mEPSCs (Fig. 1e). Thus, glucocorticoids exerted a fast inhibitory effect on glutamate release onto PVN parvocellular neurons.
Glucocorticoids inhibited glutamate release onto PVN parvocellular neurons. a, Bath application of DEX (1 μm) elicited a decrease in the frequency of mEPSCs. b, Cumulative frequency plots of mEPSC interval and amplitude distributions from the same cell showed a significant reduction in mEPSC frequency (p < 0.01) with no change in mEPSC amplitude (p = 0.28). c, Mean changes in the average mEPSC frequency, amplitude, and decay time in dexamethasone (1 μm) in 18 PVN parvocellular neurons. d, The dexamethasone-induced decrease in mEPSC frequency was dose dependent. e, Corticosterone (CORT, 1 μm) caused a similar decrease in the mean frequency of mEPSCs, whereas the corticosteroid precursor cholesterol (5 μm) and the physiologically inactive steroid isopregnanolone (5 μm) had no effect on mEPSC frequency. Numbers in parentheses represent numbers of cells analyzed in each condition in this and in the following figures. *p < 0.05; **p < 0.01.
The glucocorticoid effect is mediated by a membrane receptor
The rapid onset of the effect of glucocorticoid (3–5 min) suggested a nongenomic mechanism of steroid action. We conducted a series of experiments to determine whether the effect of glucocorticoid on glutamate release was mediated by activation of the classical intracellular corticosteroid receptors. The decrease in mEPSC frequency induced by dexamethasone was not blocked by a previous 10 min bath application of the intracellular type I or type II corticosteroid receptor antagonists spironolactone (10 μm) and mifepristone (10 μm), respectively (Fig. 2a). Additionally, dexamethasone (1 μm) applied directly into the cytoplasm of parvocellular neurons by including it in the patch pipette had no effect on the frequency of mEPSCs (101.4 ± 4.5% of baseline after 10 min of recording; n = 7) (Fig. 2a). Also, bath application of a membrane-impermeant DEX—BSA conjugate (10 μm) retained the inhibitory effect of dexamethasone on mEPSC frequency (2.18 ± 0.77 to 1.39 ± 0.54 Hz; 60.2 ± 7.3%; p < 0.01; n = 7). These data indicated that the inhibitory effect of glucocorticoids on glutamate release was not mediated by the classical intracellular corticosteroid receptors, and suggested a mechanism involving a membrane-associated receptor.
The glucocorticoid effect was mediated by a postsynaptic, G-protein-coupled membrane receptor. a, Bath application of membrane-impermeant DEX—BSA maintained the inhibitory effect, whereas intracellular application of dexamethasone (1 μm) had no effect on the frequency of mEPSCs. The intracellular glucocorticoid receptor antagonist RU486 (10 μm) and the intracellular mineralocorticoid receptor antagonist spironolactone (10 μm) failed to block the inhibitory effect of dexamethasone on mEPSCs.b, G-protein and protein-kinase blockers abolished the effect of glucocorticoid on mEPSCs. Postsynaptic G-protein blockade with intracellular GDP-β-S application (500 μm) blocked the effect of dexamethasone on mEPSCs, implicating a postsynaptic G-protein-coupled mechanism. Kinase inhibition with bath application of staurosporine (0.5 μm) or a PKC blocker, GF109203X (0.5 μm), also abolished the inhibitory effect of dexamethasone on mEPSCs, suggesting a PKC-dependent mechanism that is either presynaptic or postsynaptic. **p < 0.01.
G-protein and protein-kinase dependence
We investigated the molecular mechanisms of the rapid glucocorticoid effect in parvocellular neurons by testing for G-protein and protein-kinase dependence. The G-protein antagonist GDP-β-S (500 μm) was applied intracellularly via the patch pipette to block postsynaptic G-protein activity in the PVN parvocellular neurons. Intracellular application of GDP-β-S (10 min) blocked the decrease in mEPSC frequency evoked by dexamethasone (94.7 ± 4.7% of baseline; p = 0.11; n = 9) (Fig. 2b). Because GDP-β-S is not membrane-permeant, this indicated that the effect of glucocorticoid on glutamate release was dependent on postsynaptic G-protein activity in the parvocellular neurons. To determine the protein-kinase dependence of the glucocorticoid effect, staurosporine, a broad-spectrum protein kinase inhibitor, and GF109203X, a protein kinase C (PKC)-selective blocker, were applied in the bath 10 min before dexamethasone application. Staurosporine (0.5 μm) attenuated the inhibitory effect of dexamethasone on mEPSC frequency to non-significant levels (82.5 ± 6.9% of baseline; p = 0.25; n = 5), and GF109203X (0.5 μm) blocked the dexamethasone effect completely (102.7 ± 5.8% of baseline; n = 6) (Fig. 2b). Thus, the inhibitory effect of glucocorticoids on glutamate release in PVN parvocellular neurons is dependent on kinase activity, although it is not yet known whether the kinase dependence of the action of glucocorticoid is presynaptic and/or postsynaptic.
The glucocorticoid effect is mediated by a retrograde endocannabinoid messenger
Our data show, on one hand, that glucocorticoids reduced glutamate release, indicating a presynaptic steroid effect, and on the other hand, that the effect of glucocorticoid was dependent on postsynaptic G-protein signaling, indicating a postsynaptic site of steroid action. This suggested that the actions of glucocorticoid are mediated by the release from PVN parvocellular neurons of a retrograde messenger that inhibits presynaptic glutamate release.
NOS is expressed at high levels in the PVN (Bredt et al., 1990) and has been proposed as an intranuclear messenger in the regulation of GABA inputs to PVN magnocellular neurons (Bains and Ferguson, 1997). We tested whether NO or CO functions as the retrograde messenger that modulates glutamate release in response to glucocorticoids. Bath application of l-NAME (50 μm), a NOS inhibitor, failed to block the effect of dexamethasone (1 μm) on mEPSC frequency (decrease to 68.4 ± 6.7% of baseline; p < 0.05; n = 5) (Fig. 3a—c). Similarly, bath application of hemoglobin (20 μm), the extracellular NO and CO scavenger, also failed to block the decrease in mEPSC frequency induced by dexamethasone (1 μm) (to 72.3 ± 10.7% of baseline; p < 0.05; n = 6) (Fig. 3c). Neither CO nor NO, therefore, appears to play a role in the response to glucocorticoids.
Glucocorticoid inhibition of glutamate release was not mediated by NO or CO. a, The dexamethasone-induced suppression of mEPSCs was not blocked by the previous bath application of the NOS inhibitor l-NAME (10 μm). b, Cumulative frequency plots showed a significant reduction in mEPSC frequency (p < 0.05) with no change in mEPSC amplitude (p = 0.24) in the presence of l-NAME and DEX. c, The NO—CO scavenger hemoglobin (10 μm) also failed to block the inhibitory effect of dexamethasone on the frequency of mEPSCs. *p < 0.05; **p < 0.01.
Endocannabinoids have been shown to act as retrograde messengers that reduce synaptic glutamate and GABA release (Shen et al., 1996; Auclair et al., 2000; Wilson and Nicoll, 2001; Wilson et al., 2001). We examined whether an endocannabinoid serves as the retrograde messenger in the rapid, glucocorticoid-mediated inhibition of glutamate release. Bath application of the type I cannabinoid receptor (CB1) antagonists AM251 (1 μm) and AM281 (10 μm) blocked the dexamethasone-induced reduction in mEPSC frequency (AM251: 94.4 ± 2.7% of AM251 value, or 87.0 ± 4.9% of baseline, p = 0.20, n = 7; AM281: 99.7 ± 4.2% of AM281 value, or 82.9 ± 8.7% of baseline, p = 0.29, n = 7) (Fig. 4a,b). The type II cannabinoid receptor (CB2) antagonist AM630 failed to block the effect of dexamethasone on mEPSC frequency at a concentration selective for CB2 receptors (1 μm: 71.0 ± 9.6% of AM630 value or 69.5 ± 11.3% of baseline; p < 0.05; n = 5) but blocked the dexamethasone effect at a higher, nonselective concentration (10 μm: 101.9 ± 9.1% of baseline; n = 6) (Fig. 4c). The high-affinity synthetic cannabinoid agonist WIN55,212-2 mimicked and occluded the inhibitory effect of glucocorticoids on glutamate release. Thus, bath application of WIN55,212-2 (0.5 - 1 μm) caused a significant decrease in mEPSC frequency (70.3 ± 6.9% of baseline; p < 0.01; n = 13) but had no effect on mEPSC amplitude (101.1 ± 3.4%) or decay time (104.6 ± 2.8%) (Fig. 4d,e). Dexamethasone application (1 μm) after 10 min of WIN55,212-2 application (1 μm) failed to elicit any additional decrease in mEPSC frequency (96.2 ± 5.8% of the WIN55,212-2 value; p = 0.25; n = 5) (Fig. 4f). Therefore, the inhibitory effect of glucocorticoids on glutamate release appears to be mediated by a retrograde endocannabinoid messenger and activation of presynaptic CB1 receptors. Application of the cannabinoid antagonists AM281 (10 μm) and AM630 (10 μm) subsequent to dexamethasone application did not reverse the inhibitory effect of dexamethasone on glutamate release (data not shown), which suggested that the nonreversibility of the effect of glucocorticoid was caused by a mechanism downstream from the presynaptic cannabinoid receptors.
The effect of glucocorticoid on glutamate release was mediated by an endocannabinoid. a, The dexamethasone-induced suppression of mEPSCs was prevented by previous bath application of the CB1 receptor antagonist AM251 (1 μm). b, There was no effect of dexamethasone on the cumulative mEPSC interevent interval or amplitude distribution in the presence of AM251. c, Mean changes in average mEPSC frequency in the presence of cannabinoid receptor antagonists. The CB1 receptor antagonist AM251 blocked the effect of dexamethasone on mEPSC frequency. The CB2 receptor antagonist AM630 failed to block the DEX effect at a 1 μm concentration but blocked the DEX effect at the higher, nonselective 10 μm concentration. d, A cannabinoid receptor agonist, WIN55,212-2 (1 μm), mimicked the glucocorticoid effect, causing a selective reduction in the frequency of mEPSCs. e, Cumulative frequency plots showed a significant increase in mEPSC interval (p < 0.01) but no change in mEPSC amplitude (p = 0.42) in WIN55,212-2. f, Mean changes in the average mEPSC frequency in the presence of glucocorticoid and the cannabinoid receptor agonist. The effect of WIN55,212-2 was similar to that of dexamethasone, but these effects were not additive, suggesting converging mechanisms. *p < 0.05; **p < 0.01.
Glucocorticoid effects on multiple parvocellular cell types
Parvocellular PVN neurons tested for their sensitivity to glucocorticoids or cannabinoids were screened for CRH, TRH, OT, and VP mRNA expression using the single-cell RT-PCR technique after experiments (Surmeier et al., 1996; Glasgow et al., 1999). Of 30 neurons responsive to glucocorticoids, five expressed CRH, two expressed TRH, four expressed OT, and three expressed VP mRNAs. Of 16 neurons responsive to cannabinoid analogs, one expressed CRH, four expressed OT, and two expressed VP mRNAs. Each of the identified cells expressed only one of the four mRNAs (Fig. 5), except one cell in which expression of both TRH and OT was detected. These findings indicate that the glucocorticoid-induced suppression of glutamate synaptic inputs occurs in different peptidergic phenotypes among neurosecretory parvocellular PVN neurons, and suggest a generalized feedback inhibitory role of glucocorticoids in the regulation of hypophysiotropic hormone secretion.
RT-PCR analysis in individual parvocellular neurons. a, CRH mRNA expression in six individual PVN parvocellular neurons. The expected CRH PCR product is 326 bp. b, TRH, VP, OT, and GAPDH mRNA expression in individual PVN parvocellular neurons. The expected PCR product weights are 276 bp (TRH), 440 bp (VP), 463 bp (OT) and 441 bp (GAPDH). The cell identification numbers are shown above each lane. A DNA ladder ranging from 300 to 500 bp is shown in the left lane of each panel.
Together, our data demonstrate a fast, membrane corticosteroid receptor-mediated suppression of excitatory input to neurosecretory parvocellular neurons of the PVN, and support a model of acute glucocorticoid actions (Fig. 6). In this model, glucocorticoids activate a postsynaptic G-protein-coupled membrane receptor that leads to the release of an endocannabinoid. The endocannabinoid acts as a retrograde messenger at presynaptic glutamate terminals to inhibit glutamate release onto the parvocellular neurons, thereby decreasing the excitatory synaptic drive to these cells.
A model of the rapid glucocorticoid actions in PVN parvocellular neurons. The proposed mechanisms include glucocorticoid (CORT) binding to a G-protein-coupled glucocorticoid receptor (GRmb) and activation of an intracellular signaling pathway (dashed arrow) that leads to endocannabinoid synthesis (+). Endocannabinoid (CB) is released from the PVN neuron and binds to a G-protein-coupled CB1 cannabinoid receptor (CBR) on presynaptic glutamate terminals, activating a signaling cascade (dashed arrow) that leads to the inhibition of glutamate (GLU) release onto the PVN neuron-leading to decreased PVN neuronal activity and hormone secretion.
A membrane G-protein-coupled glucocorticoid receptor
Several findings presented here support the involvement of a membrane glucocorticoid receptor in the acute effects of corticosteroids on PVN parvocellular neurons: (1) the rapid response to extracellular corticosteroids, (2) the lack of effect of intracellular corticosteroid application, (3) the maintained effectiveness of the membrane-impermeant DEX—BSA conjugate, and (4) the failure of antagonists of the classical intracellular corticosteroid receptors to block the acute corticosteroid effect. Additionally, blockade of the corticosteroid effect by blocking G-protein activity selectively in the postsynaptic neuron implicates a G-protein-signaling mechanism and suggests that the membrane corticosteroid receptor in PVN parvocellular neurons is a G-protein-coupled receptor. Whether the kinase dependence of the response is associated with the postsynaptic G-protein signaling that leads to the release of endocannabinoid or with the presynaptic cannabinoid receptor signaling remains to be determined. These findings are consistent with reports of G-protein and/or protein-kinase involvement in rapid corticosteroid actions in the brain, including radiolabeled corticosterone binding in amphibian neuronal membranes (Orchinik et al., 1992) and cortisol modulation of calcium currents in guinea pig CA1 pyramidal neurons (ffrench-Mullen, 1995).
Release of a retrograde endocannabinoid messenger
We found that the activation of a postsynaptic, G-protein-dependent signaling mechanism led to the suppression of glutamate release, a presynaptic endpoint, thus implicating the involvement of a retrograde messenger. Inhibiting the retrograde messengers NO and CO failed to block the effect of glucocorticoid on mEPSC frequency or inhibition of cannabinoid receptors. Conversely, saturation with exogenous agonist or antagonist, respectively, abolished the effect of glucocorticoid on mEPSCs completely, suggesting that the retrograde messenger involved is an endocannabinoid. Our findings indicate that the glucocorticoid-induced endocannabinoid actions were mediated by presynaptic CB1 receptors, because they were blocked by CB1 receptor antagonists as well as by a CB2 receptor antagonist at a concentration effective at both CB2 and CB1 receptors but not at a concentration selective for CB2 receptors. This is consistent with the finding that CB1 receptors are the predominant receptors in the brain (Matsuda et al., 1990; Munro et al., 1993). Retrograde regulation of the synaptic release of glutamate and GABA by endocannabinoids is emerging as a widespread modulatory mechanism at synapses throughout the brain, including the hippocampus, cerebellum, striatum, and neocortex (Huang et al., 2001; Ohno-Shosaku et al., 2001). It is worth noting that the CB receptor antagonists we used are considered inverse agonists (Gifford et al., 1997; Ross et al., 1999), although neither had opposite actions on glutamate release, suggesting that they may not play an inverse agonistic role at the presynaptic receptors in the PVN at the concentrations used. Future studies will clarify the CB receptor mechanisms in the PVN.
Introduction of GDP-β-S into a single parvocellular neuron blocked the actions of glucocorticoids completely, suggesting that there was little or no spillover of endocannabinoids released by adjacent neurons onto the recorded neurons. This is consistent with a restricted secretion domain and limited spread of the endocannabinoids in the extracellular space. Unlike the gaseous retrograde messenger NO, which has been shown to spread to activate local GABA circuits in the PVN (Bains and Ferguson, 1997), retrogradely released endocannabinoids might be expected to have a restricted range of action because of their lipophilic nature (Kreitzer and Regehr, 2002).
It is interesting to note that the inhibitory actions on PVN neurosecretory cell activity of glucocorticoids and cannabinoids applied directly in the PVN in our experiments are opposite to the activational effects of systemic anandamide or Δ 9-tetrahydrocannabinol on HPA hormone release (Murphy et al., 1998). However, the effects of systemic cannabinoids on HPA hormone secretion are accompanied by changes in PVN catecholamine levels (Murphy et al., 1990; Rodriguez de Fonseca et al., 1995), and are blocked completely by hypothalamic deafferentation (Puder et al., 1982). This suggests that the activational effects of cannabinoids occur upstream of the PVN, and that this cannabinoid-stimulated afferent excitation either is not glutamatergic or is glutamatergic and strong enough to override the CB1 receptor-mediated inhibition of glutamate release within the PVN.
Physiological significance
Glucocorticoid feedback inhibition of the HPA axis appears to be mediated by steroid actions at different central targets, either indirectly via actions in the hippocampus or directly at the levels of the hypothalamus and pituitary (Dallman et al., 1987). Glucocorticoids have been reported to have direct inhibitory effects on the electrical activity of some parvocellular neurons of the PVN (Kasai and Yamashita, 1988; Saphier and Feldman, 1988), although the inhibitory mechanisms have not been determined and the identification of specific glucocorticoid-responsive PVN subpopulations has been inconclusive. We present evidence here for the inhibition by glucocorticoids of glutamate release onto parvocellular neurosecretory neurons of the PVN, including CRH-expressing cells, which resulted in the suppression of excitatory inputs to these cells. This is the first demonstration, therefore, of a rapid inhibition of identified CRH neurons by corticosteroids, and provides a mechanism for the fast feedback inhibition of CRH release by glucocorticoids directly at the level of the hypothalamic CRH neurons.
In addition to the effect on CRH neurons, we found that glucocorticoids also exerted a similar inhibitory effect on glutamate release onto parvocellular TRH-expressing, OT-expressing, and VP-expressing neurons. These peptides, along with CRH, are thought to play an anorexigenic role in the central control of feeding (Burlet et al., 1992; Rondeel et al., 1992). Glucocorticoids and endocannabinoids both have well established central orexigenic actions (Zakrzewska et al., 1999; Kirkham et al., 2002) such that the modulatory cross talk between glucocorticoids and endocannabinoids that leads to reduced excitation of PVN parvocellular neurons demonstrated here may also play a major role in the central control of feeding and energy balance.
The glucocorticoid—endocannabinoid interface via activation of a G-protein-dependent signaling pathway represents a novel mechanism of rapid steroid action in the brain. This endocannabinoid link between steroid hormonal and neuronal signaling expands the recently discovered role of endocannabinoids as retrograde modulators of synaptic function. The glucocorticoid-induced suppression of excitatory synaptic inputs to PVN parvocellular neurons is likely to play an important role in the acute negative feedback effect of glucocorticoids on HPA axis activity. Because the PVN is considered to be the main neuroendocrine and autonomic effector in the control of stress, energy balance, and homeostasis (Swanson and Sawchenko, 1980; Laugero, 2001), the modulatory cross talk between glucocorticoids and endocannabinoids in PVN parvocellular neurons provides a central mechanism for the well established orexigenic and homeostatic actions of both glucocorticoids and cannabinoids. This novel role of endocannabinoids as mediators of the negative feedback actions of glucocorticoids in the PVN provides new potential targets for the therapeutic treatment of stress and feeding disorders.
This work was supported by National Institutes of Health Grants NS/DK39099 and MH066958. We thank Harold Gainer, Eric Glasgow, and D. James Surmeier and his laboratory colleagues for their help in setting up the single-cell RT-PCR technique in our laboratory. We thank James Herman and F. Edward Dudek for their critical reading of this manuscript.
Correspondence should be addressed to Dr. Jeffrey G. Tasker, Department of Cell and Molecular Biology, 2000 Percival Stern Hall, Tulane University, New Orleans, LA 70118-5698. E-mail: tasker{at}tulane.edu.
Copyright © 2003 Society for Neuroscience 0270-6474/03/234850-08$15.00/0
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Journal of Neuroscience
Vol. 23, Issue 12
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Journal of Neuroscience 15 June 2003, 23 (12) 4850-4857; DOI: 10.1523/JNEUROSCI.23-12-04850.2003
mTORC2 Loss in Oligodendrocyte Progenitor Cells Results in Regional Hypomyelination in the Central Nervous System
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text:
Q: how to make embed in navigation controller work with scrollView I have watched this tutorial
Snapchat Like Layout using View Controller Theme
then I embed one of the three controllers in Navigation Controller but it didn't work then i have tried to add it on the view controller how had the scroll view also didn't work + gives me overlap I didn't tried it in code tho but I don't think this is the cause , this is the code

class ViewController: UIViewController {

    @IBOutlet weak var scrollView: UIScrollView!
    override func viewDidLoad() {
        super.viewDidLoad()

        let V1 = self.storyboard?.instantiateViewController(withIdentifier: "V1") as UIViewController!
        self.addChildViewController(V1!)
        self.scrollView.addSubview((V1?.view)!)
        V1?.didMove(toParentViewController: self)
        V1?.view.frame = scrollView.bounds

        let V2 = self.storyboard?.instantiateViewController(withIdentifier: "V2") as UIViewController!
        self.addChildViewController(V2!)
        self.scrollView.addSubview((V2?.view)!)
        V2?.didMove(toParentViewController: self)
        V2?.view.frame = scrollView.bounds

        var V2Frame: CGRect = V2!.view.frame
        V2Frame.origin.x =  self.view.frame.width
        V2?.view.frame = V2Frame

        let V3 = self.storyboard?.instantiateViewController(withIdentifier: "V3") as UIViewController!
        self.addChildViewController(V3!)
        self.scrollView.addSubview((V3?.view)!)
        V3?.didMove(toParentViewController: self)
        V3?.view.frame = scrollView.bounds

        var V3Frame: CGRect = V3!.view.frame
        V3Frame.origin.x = 2 * self.view.frame.width
        V3?.view.frame = V3Frame

        self.scrollView.contentSize = CGSize(width: self.view.frame.width * 3, height: self.view.frame.height)
        self.scrollView.contentOffset = CGPoint(x: self.view.frame.width * 1 , y: self.view.frame.height)

    }

how the code works ??
1- first I have added scroll view to a view controller
2- then inside the scroll view I have added 3 view controller and all the 3 controllers has own class like normal view controllers
3- the scroll View added the 3 controllers on its view by this code on the top
all the 3 controllers inheriting from own class and everything is wor fine
if anyone can help or need more description just ask

A: try below

Create ViewController in Storyboard or XIB and add ScrollView and PageControl

My Class as below:

class ScrollContentViewController: UIViewController, UIScrollViewDelegate {

    @IBOutlet weak var pageControl: UIPageControl!
    @IBOutlet weak var scrollView: UIScrollView!
    override func viewDidLoad() {
        super.viewDidLoad()

        // Do any additional setup after loading the view.
    }

    override func viewDidAppear(_ animated: Bool) {
        super.viewDidAppear(animated)
        self.pageControl.currentPage = 0
        self.pageControl.numberOfPages = 4
        var originX = CGFloat(0)
        let width = scrollView.frame.size.width
        let height = scrollView.bounds.size.height
        for x in 0 ..< 4 {
            let V1 = UIViewController()
            self.scrollView.addSubview(V1.view)
            V1.view.frame = CGRect(x: originX, y: 0, width: width, height: height)
            V1.view.backgroundColor = x % 2 == 0 ? UIColor.lightGray : UIColor.darkGray
            self.addChildViewController(V1)
            originX += width
        }
        self.scrollView.contentSize = CGSize(width: originX, height: height)
    }

    override func didReceiveMemoryWarning() {
        super.didReceiveMemoryWarning()
        // Dispose of any resources that can be recreated.
    }

    func scrollViewDidEndDecelerating(_ scrollView: UIScrollView) {
        let x = Int(scrollView.contentOffset.x / scrollView.frame.size.width)
        self.pageControl.currentPage = x
    }

    /*
    // MARK: - Navigation

    // In a storyboard-based application, you will often want to do a little preparation before navigation
    override func prepare(for segue: UIStoryboardSegue, sender: Any?) {
        // Get the new view controller using segue.destinationViewController.
        // Pass the selected object to the new view controller.
    }
    */

}

redpajama_set_name: RedPajamaStackExchange


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There's space to record the wine colour (red, white or rosé) as well as where you bought it and for how much. It's also useful to keep a note of what the bottle label says so that if you like the wine you'll have a good chance of finding exactly the same, or a similar type, again.
The tasting sheet also has space to record the grape varieties used in the wine – these will be displayed on the label, unless it's French, when these will be hidden from you, because you shouldn't need to ask should you? A white Burgundy will be made from Chardonnay grapes, a red Burgundy from Pinot Noir, and Bordeaux reds (or as we English like to call them, Clarets), will be a blend of Merlot and Cabernet Sauvignon grapes.
Once you've written all that down you'll finally pour the wine. Give it a sniff so you can make a note of its aroma and then sip and taste. There are no right or wrong words for the aroma and taste – write down whatever you think the wine smells and tastes of even if that's home-made custard or Persil washing up liquid.
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The USMNT was fortunate to walk out of Houston with a 1-1 draw against Chile. While the two-time defending Copa America champions had a better game, there were positives to take away for Gregg Berhalter. ASN's Brian Sciaretta looks at how the U.S. players performed with his ratings.
THE UNITED STATES national team played to a 1-1 draw with Chile on Tuesday night in a game that was an important learning experience for Gregg Berhalter's team. Overall the result was fortunate for the U.S. team but there was still plenty that went right for the Americans.
The United States got off to a strong start and scored first on a nice play from Christian Pulisic that was set up by Gyasi Zardes. The lead only lasted five minutes as shambolic defending from the United States eventually gave way to Chile's equalizer.
After an awful end to the first half and start to the second half, the U.S. team eventually grew back into the game and had a few chances to press for a goal. Playing without Tyler Adams, Weston McKennie, and Christian Pulisic from the 36th minute onward, the U.S. team will have more weapons moving forward. In their place, Berhalter had the chance to evaluate several other players.
What helped the U.S. team was some tactical adjustments as a 5-4-1 formation in the second half substantially reduced Chile's opportunities.
In the end, neither team was able to find a winner. For Berhalter, there will be plenty to digest from this game moving forward into the Gold Cup.


The US Transportation Security Administration has approved direct flights from the Ivory Coast to the United States.
The country aims to become a hub for the region and hopes the new routes will help to attract more US investors to the region. The Ivory Coast has been seeking authorization from the US for such routes for 15 years.


text:
Ravens 2019 Post-Draft Roundtable
A week ago today, the Ravens made their first pick in the 2019 draft, trading back from 22 to 25 to select Marquise "Hollywood" Brown, wide receiver, Oklahoma. The first pick of Eric DeCosta's tenure. BSL's Mike Randall, Gabe Ferguson, Chris Stoner, and Rivers McCown, discuss their thoughts on how draft weekend played out, and some outlook for the upcoming season.
Discuss your thoughts on the topics we've posted here on our message board. Let's dive right in.
1. What are your thoughts on the events that led up to the Ravens drafting WR, Marquise Brown, with the 25th overall pick? Was there another player you thought would be there and wasn't? Was trading back the right move?
Mike Randall – I think trading back was the right move. Could have been happy with any number of three or four guys who were still available at 22, picked up some picks in the process. Garrett Bradbury would have bolstered the O-line in this run-heavy attack we're going to see in 2019. But since he was gone shortly before 22, I'm ok with trading back and still getting an electrifying player in Brown and becoming likely the fastest offense in the league.
Gabe Ferguson – Definitely liked trading back. I don't think there was anyone on the board at 22 that was a run to the podium type selection. I think I would have preferred drafting Montez Sweat at 25, but it is impossible to know how the Ravens felt about him from a medical standpoint. In the end I think Brown is an exciting player for the Ravens offense and a solid selection at that point in the draft.
Chris Stoner – I feel the Ravens have typically allowed the board to fall to them. They came in with multiple needs, meaning there were several ways they could have gone. Clearly with their trade falling back, they made the calculation that someone they valued highly (including Brown) would still be available. I was excited by the trade and picking up additional picks.
Rivers McCown – It's hard to ever knock a team for trading back, and because the Ravens were blessed with two solid bookended tackles and a draft slot right ahead of a team that was absolutely desperate for tackles, it was a prime spot to trade out of. I don't think the Ravens missed out on anybody important to their team philosophy by making the trade.
2. With the "Hollywood" pick, then doubling up with another wideout in Miles Boykin in round three, what are your thoughts on the receiver group as a whole?
MR – In one word, skeptical. I'll come out and say it. I'm not a fan of the Boykin pick. The guy gets zero separation from DBs, but he does have a pro-looking body type and will block. Curious how they will use Brown. Contested catches was not his forte and he won't be running by defenses like they were standing still like he could in the Big 12. I like Willie Snead. I feel like Chris Moore should battle for more playing time this season. Indifferent on Seth Roberts. Feel like Jordan Lasley and Jaleel Scott are cast offs, leftovers from a previous era. I think the most important thing will be lack of selfishness. The top receivers on this team might get three or less targets some games.
GF – It went from being a bottom 3 unit in the league to at least middle of the pack. We will obviously have to wait and see how these rookies pan out, but both Brown and Boykin were very highly regarded and bring highly complementary skill-sets. It's clear that DeCosta wanted to surround his young QB with weapons and he did just that.
CS – The Ravens could have selected Brown, patted themselves on the back and celebrated. I like that that made the decision that Brown by himself wasn't enough. Boykin represents a different skill set, and to me that is a key. You want a group of varied skills and abilities. You also want depth and usable options when inevitable injuries occur. Realistically they are going to keep 5 or 6 WR's. Brown, Snead, Boykins, Moore, Lasley, Roberts. With the two picks, it looks a lot better. It's still not great. It's two rookies, and I'm sure they will both flash, but how consistent will they be? Snead is usable. Moore a good ST player. Maybe this is the year his WR production jumps? Lasley has talent but doesn't matter if you don't have focus. Let's see what he looks like in Y2. I like that Roberts has missed two games in four years and made at-least 32 grabs per year in his career.
If we're talking 'pass catching' group as a whole, the RB's are going to get their share, and the TE's are obviously going to get a lot of use. When you factor in those groups, I feel good overall.
RM – Hollywood is a more-than-ample John Brown replacement. It's kind of interesting to me that the Ravens seemed to zig towards Greg Roman's run game so hard in free agency, then zag back towards a passing game in the draft class. I think they got a pair of super-athletic mismatches that will be pressed into duty as soon as they can. That's not a great indicator of short-term success — especially with Brown coming off a Lisfranc injury — but long-term I like it.
3. The Ravens second pick, first pick in round three, was edge rusher Jaylon Ferguson. Thoughts on the pick here, where does he fit on your depth chart?
MR – Tim Williams is put on notice with this pick. Should be a battle between he and Ferguson in camp to take Terrell Suggs spot on the edge, with Matt Judon starting opposite them. Solid pick. I don't put stock into college stats. But Ferguson was the all-time leader in sacks at the college level. Terrell Suggs was the college single season sacks record holder, and despite poor athletic numbers, he did all right for himself. I think the pick will work out just fine.
GF – Best pick of the draft in my opinion, and not just because he has a great name. Ferguson is a natural pass-rusher with immense college production, and he still has room to improve. He was talked about as a possible 1st round selection at one point so you have to love the value too. In terms of the depth chart, Edge rusher was probably one of the biggest needs on the team, so I see him coming in and competing with Tim Williams and Tyus Bowser to start at OLB across from Matthew Judon.
CS – Ferguson was who I wanted there. The knocks I've read on him are that he hasn't shown Michael McCrary's constant motor, and he isn't a freak athlete. What I like is the production. When he was selected, on the stage was Adalius Thomas – who like Ferguson – was a former Conference USA Defensive Player of the Year. And Thomas was the first one I thought of with Ferguson. Thomas was a plus athlete, with his ability to drop in coverage, and work on Special Teams. That might not be Ferguson, but the comparison is that they both did work between the lines and were a bit overlooked. I'd give Ferguson a chance to win the starting job at DE, and I'd be showing him Suggs highlights on setting the edge against the run. My best guess is that he'll mostly be used in passing downs and sent on seek and destroy missions.
RM – Ferguson seemed to be someone who divided scouts. He had a ton of production as the FBS career sack leader, but he didn't test very well athletically and was uninvited from the combine based on a (admittedly minor) battery charge. At the end of round 3, I think you'd be hard-pressed to do better. Ferguson probably won't start Game 1 but I could see him breaking into the lineup fairly quickly, perhaps by midseason.
4. On to day three. Rounds four through seven. First, let's hit on Trace McSorley, QB out of Penn State. Are you buying the hype that he is going to be our version of the Saints Taysom Hill? Is that as much of an X-factor as people make it out to be?
MR – No, and no. Lamar Jackson was a different look when he was out there with Joe Flacco. Two different styles. You try to catch the defense in a mismatch. McSorley out there with Lamar Jackson is just another athlete on the field, when the defense has already game-planned to try to slow down the litany of athletes the Ravens now possess. Not really an X-factor, in my opinion. Something way too overrated in the game. Wasn't game changing when Jackson saw the field with Flacco. Isn't game changing when the Saints have Drew Brees and Taysom Hill share the field.
GF – I liked the pick. Everything I have read and heard about McSorley is very positive; leader, hard worker, competitor. Sounds like the kind of guy the Ravens love. In terms of his role on the team, I wouldn't be surprised to see him used in some packages on offense, but also playing some ST and maybe even reserve DB.
CS – If the Ravens have to rely on McSorley to QB, they'll be in a world of hurt; but at-least they could run similar concepts to what Jackson and RG3 will be doing. I like the Hill comparison. They could keep all QB's active, use McSorley as the gimmick guy, and use RG3 to start if Jackson gets hurt. I'd feel better about McSorley being on the roster, if he shows in camp that he could capably fill in at either WR or CB.
RM – I think McSorley was a wasted pick. The Hill experiment is basically all about the idea that NFL teams get accustomed to defending certain gaps and that a more mobile quarterback changes things and catches them off-guard. That matters when Drew Brees is your quarterback. Not really so much when Lamar Jackson is your quarterback.
I'm not gonna blame Eric DeCosta for the pick because I think most low-round picks will wash out anyway, but it wouldn't be where I'd spend a pick.
5. Outside of McSorley, which day three pick do you feel has the potential to be a major contributor on this team?
MR – Ben Powers, G, Oklahoma. Made 35 career starts in college. First Team All-American. Looks like he could be Brock Lesnar's younger brother. Other than getting pushed around by Quinnen Williams (who didn't get pushed around by him?) he looks like a solid guard and should challenge Alex Lewis for the LG gig. Light on his feet, good pulling guard, likely to be mentored by Marshal Yanda. What isn't to like there?
GF – Probably Ben Powers, OG from Oklahoma. I think he will compete to start as a rookie at LG. Justice Hill should also carve out a niche role at RB, but it's hard to see him seeing a ton of work as a rookie. A larger role will likely develop for him down the road.
CS – For 2019, I think the answer is Justice Hill. The Day 3 pick that feels very 'Ravens' is Daylon Mack.
RM – Ben Powers sort of got lost in the shuffle at OU, since OU had three linemen drafted and Powers didn't have the speed to stay outside. I like his technique and think he will do a major service for this team pushing Alex Lewis and perhaps eventually becoming Marshal Yanda's stand-in.
6. It's early, but after the draft, many positions are overloaded, especially as teams boost the roster to 90 guys with undrafted free agents. Which position battle are you looking forward to this summer?
MR – I mentions WR, Edge, LG already. Another one might be tight end. Not over loaded. But I want to see if Hayden Hurst blossoms into a special player as the first-round pick of 2018. Mark Andrews was the third-round pick. But if you didn't know any better and based on last year, you'd swear the picks were reversed.
GF – OLB will be fun to watch. The Ravens have one definitive starter, and three guys who could all lay claim to a big role in the defense. In the end I think we will see all four players in a rotation, but it will be telling to see who is out there most consistently on 3rd down.
CS – The secondary is loaded. Iman Marshall just adds to that. There is a battle for the 5th WR, and for the '6th' to show the Front Office and Coaching staff they also need to be included on the roster.
Should be a battle at Center, and for spots 6 to 8 on the O-Line as a whole.
RM – Running back is going to be crucial — I think Justice Hill has a chance to be a great passing-down back and Mark Ingram, obviously, got signed to be a big piece of the puzzle. So, I'm very interested to see where this leaves the Gus Bus — he did so well last season, is he nothing more than depth at this point? Would another team take a chance on him? Could be a curious offseason for him.
7. if you look at the Ravens draft history, more often than not you can pick out three players from each class who were major contributors during their tenure in Baltimore. Which three guys of the eight drafted, and 19 undrafted if you like, will wind up being the cream of the 2019 crop when we get to say, 2023?
MR – I think Marquise Brown at worst ends up like Tavon Austin. Austin is still in the league playing a role in Dallas. Jaylon Ferguson could be a steal in round three. Ben Powers would be my third player I expect to contribute nicely for a rookie contract length of time.
GF – Brown, Ferguson and Powers would be my guess for the biggest contributors, but I think Boykin will have a pretty big role in this offense as well.
CS – Ferguson, Hill, Mack. I was pleased that the Ravens were able to get Gerald Willis as an UDFA.
RM – Brown, Ferguson, Hill.
8. Finally, this was Eric DeCosta's first draft as Ravens GM. I don't want to say "give DeCosta a grade" because grades at this point are irrelevant. But, what did you think?
MR – Looked like an Ozzie Newsome draft at first with the trade back. They looked to have a mission of adding even more speed to the offense, and replacing Terrell Suggs, if you even can. I liked some other guys around the time of the Boykin pick, but I could be wrong. Been wrong before. Will be wrong again at some point. All I know is if Lamar Jackson to Marquise Brown turns into Pat Mahomes to Tyreek Hill, DeCosta will have done more for the WR position through the draft than Ozzie ever did. Though Brown and Ferguson will have to be enshrined in Canton one day for DeCosta to achieve master status, like Ozzie did with his first two picks.
GF – I'll give DeCosta a grade and he'll get an A- from me. My only complaints would be waiting a bit longer than I would like to draft offensive line and taking Brown over Sweat in round one. Aside from that I think he knocked his draft out of the park.
CS – I thought DeCosta did an excellent job. We won't know how good the class is for a while, but the move from 22 to 25 was fantastic. Adding multiple quality WR's was great. The Ravens came out of the draft, still having questions at Center and with depth at ILB; but beyond that – the roster is in very good shape. The team is younger, faster, and more athletic than it has been in sometime. There is room to grow. This could be a good year, it boils down to Jackson's development. I like what DeCosta was able to achieve here in the draft, and what he's done overall this off-season.
RM – If you pasted this Ravens draft into 2013, I don't think anyone would notice Ozzie Newsome wasn't the general manager. That's a compliment.
Written by Mike Randall. Posted 3 months ago in Baltimore Ravens.
Mike Randall
Mike was born on the Eastern Shore, raised in Finksburg, and currently resides in Parkville. In 2009, Mike graduated from the Broadcasting Institute of Maryland. Mike became a Baltimore City Fire Fighter in late 2010. Mike has appeared as a guest on Q1370, and FOX45. Now a Sr. Ravens Analyst for BSL, he can be reached at [email protected]

redpajama_set_name: RedPajamaCommonCrawl


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Excavation at Khirbet Qeiyafa
Photo: Sky View, courtesy of Hebrew University and Israel Antiquities Authority
Archaeologists unveil King David's palace
Royal storerooms also revealed in joint excavation of Hebrew University, Israel Antiquities Authority at Khirbet Qeiyafa, a fortified city in Judah identified with biblical city of Shaarayim
Ynetnews|Published: 07.19.13 , 07:55
Two royal public buildings, the likes of which have not previously been found in the Kingdom of Judah of the 10th century BCE, were uncovered this past year by researchers of the Hebrew University and the Israel Antiquities Authority at Khirbet Qeiyafa – a fortified city in Judah dating to the time of King David and identified with the biblical city of Shaarayim.
One of the buildings is identified by the researchers – Prof. Yossi Garfinkel of the Hebrew University and Sa'ar Ganor of the Israel Antiquities Authority – as David's palace, and the other structure served as an enormous royal storeroom.
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The excavation, which was conducted over the past seven years, is now drawing to a close. According to Prof. Garfinkel and Sa'ar Ganor, "Khirbet Qeiyafa is the best example exposed to date of a fortified city from the time of King David. The southern part of a large palace that extended across an area of around 1,000 square meters (10,700 square feet) was revealed at the top of the city.
"The wall enclosing the palace is about 30 meters (100 feet) long, and an impressive entrance is fixed it through which one descended to the southern gate of the city, opposite the Valley of Elah. Around the palace's perimeter were rooms in which various installations were found – evidence of a metal industry, special pottery vessels and fragments of alabaster vessels that were imported from Egypt.
Aerial picture of site (Photo: Sky View, courtesy of Hebrew University and Israel Antiquities Authority)
"The palace is located in the center of the site and controls all of the houses lower than it in the city. From here one has an excellent vantage looking out into the distance, from as far as the Mediterranean Sea in the west to the Hebron Mountains and Jerusalem in the east. This is an ideal location from which to send messages by means of fire signals. Unfortunately, much of this palace was destroyed 1,400 years later when a fortified farmhouse was built there in the Byzantine period."
A pillared building about 15 meters (49 feet) long by 6 meters (20 feet) wide was exposed in the north of the city, which was used as an administrative storeroom.
According to the researchers, "It was in this building the kingdom stored taxes it received in the form of agricultural produce collected from the residents of the different villages in the Judean Shephelah. Hundreds of large store jars were found at the site whose handles were stamped with an official seal as was customary in the Kingdom of Judah for centuries."
Finds from site (Photo: Clara Amit, courtesy of Israel Antiquities Authority)
The palace and storerooms are evidence of state sponsored construction and an administrative organization during King David's reign.
"This is unequivocal evidence of a kingdom's existence, which knew to establish administrative centers at strategic points", the archaeologists say. "To date no palaces have been found that can clearly be ascribed to the early tenth century BCE as we can do now. Khirbet Qeiyafa was probably destroyed in one of the battles that were fought against the Philistines circa 980 BCE.
"The palace that is now being revealed and the fortified city that was uncovered in recent years are another tier in understanding the beginning of the Kingdom of Judah."
The exposure of the biblical city at Khirbet Qeiyafa and the importance of the finds discovered there have led the Israel Antiquities Authority to act together with the Nature and Parks Authority and the planning agencies to cancel the intended construction of a new neighborhood nearby and to promote declaring the area around the site a national park.
This plan stems from the belief that the site will quickly become a place that will attract large numbers of visitors who will be greatly interested in it, and from it one will be able to learn about the culture of the country at the time of King David.
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Photo by Teresa Kamakea used with permission
1st Tesla Owner To Use Tesla's New, Gigantic V3 Supercharger Station In Firebaugh Shares Her Story
Tesla just installed a total of 56 of its new V3 superchargers in Firebaugh, California, along Interstate 5, which is the main stretch for West Coast drivers traveling between San Francisco and LA.
Tesla just installed a total of 56 of its new V3 superchargers in Firebaugh, California, along Interstate 5, which is the main stretch for West Coast drivers traveling between San Francisco and LA. In a conversation with the Tesla Owners of East Bay account on Twitter, I learned that the Kettleman City supercharger station is around an hour south of the new station and has 40 chargers. Three years ago, it was the biggest Tesla Supercharger station in North America. Teresa Kamakea, one of the members of the Tesla Owners of East Bay, was the very first customer to ever use the brand new supercharger. She told me in an email that both she and her daughter agreed that it was kismet, for sure.
Seeing A Unicorn
Teresa was thrilled to even run across a V3 supercharger for the first time. "Charging at a V3 charger for the first time was like seeing a unicorn to me. I belong to Tesla Owners East Bay, and the club always keeps us up to date on all that is new and exciting in Tesla, so I had been reading a lot of posts about them on our club Facebook page and was hyped up about it. But the first time I'd seen a V3 charger was last night at the Kettleman City Supercharger — not the new Firebaugh Superchargers."
"I stopped there on my way home from helping my elderly father in Southern California. They have clearly labeled all of their 150kW and 250kW chargers and I was freaking excited because I didn't know they had the V3 chargers there — they weren't there the last time I went through. I got charged to 306 miles on my battery in about 30 minutes while I was walking my dog and then browsing the Tesla gear and using the bathroom myself in the lounge. That was more than enough charge to get me all the way home to Livermore without needing to stop again. That was the first time that I'd used the V3 charger."
Teresa's Experience At The New Firebaugh Superchargers
Teresa's story of how she found these new Superchargers seems like something out of a fictitious adventure. You're running an errand or something and you need to stop for a minute or are suddenly forced to take a detour, and then you find something. It's equivalent to finding a hidden bookstore off to the side in an area you probably wouldn't go in because it's never crossed your mind — as she said earlier, kismet.
"So when I pulled into the brand new Firebaugh location just 60 miles down the road, it was not because I needed a charge. My doggie was acting like she had to go, so I was just pulling off at the first exit that looked safe, Panoche Rd exit. That is when I saw the new Tesla superchargers at Firebaugh, but I didn't know that's what it was. It wasn't on my Supercharger map and I was kinda shocked because in all of my trips I'd never seen it before and you can't miss it if you're on 5. I was freakin excited to see from a distance that there were so many chargers and they all had skinny cables!!! That's how I can tell the V3 from the V2 chargers — the size of the cable. Size matters. 😀 "
Teresa's 1st Experience Supercharging Compared with 1st Experience V3 Charging
I asked Teresa to give me a comparison so we could show how far along Tesla has come with its Supercharging network. Teresa's very first Supercharger experience was at the Bakersfield Superchargers. "I remember that my screen said it would take 45 minutes to charge enough to get the rest of the way to my dad's house in Temecula. That really was not enough time for me to walk my dog, go to the bathroom myself, eat, and visit with the other Tesla owners that I met that were also charging there. I didn't note or calculate how long it took to charge per kW."
She noted that the charging speed has never been a concern, with her pointing out that, obviously, she doesn't want to have to wait hours to charge, but even the older V2 superchargers would still finish charging before she was finished with her break from a long 8–10 hour drive.
She shared her thoughts about progress with me as well. "For me, the progress I've noticed is in the feel of the supercharger stations. At Bakersfield where I used a Supercharger for the first time, the chargers are a minority on the huge lot in comparison to all of the space that the gas pumps take up. It feels like, well, you can have a little bit of unused space here in the corner of our lot for your chargers. But the Firebaugh Supercharger station like the Kettleman City Supercharger station makes me feel like we belong and we are here to stay."
"I've always thought that if I want to know the exact numbers, it would be easy enough to look that up to calculate the difference and progress from V2 to V3 chargers. For me, the more important progress is in the growth and acceptance of Tesla."
Looking at Teresa's photos that she provided for the article, I noticed something. The new V3 supercharger has both a Chevron and a Shell gas station within sight. This is brilliant, and strategic marketing by Tesla. Having a large Supercharging station catty-corner two gas stations off of a major interstate exit should be the norm for Tesla.
Looking at these photos is like seeing the future in front of you while frozen in time. One day, these gas stations will be either closed or converted into something else. As we observe this frozen moment while still in the present, it gives me an incredible sense of awe to be able to note this. One day, we will look back at this article and Teresa will say, "I remember when those gas stations were there. Now there's something else."
And that something else will most likely be related to Tesla or electric vehicles.
All photos by Teresa Kamakea used with permission. Featured photo by Teresa and edited by Johnna for artistic effects. The last thing the fossil fuel industry will see before it dies is the red glow of a Tesla Supercharger.
In this article:California, Tesla, tesla superchargers, Tesla Supercharging, Tesla V3 Supercharging
If all goes according to plan, mobile hydrogen fueling stations will solve the chicken-and-egg conundrum for fuel cell trucks in USA.
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Walmart is testing the waters of cashier automation, first by letting customers scan and pay for items within an app, and now, giving in-store reps the ability to help customers pay on mobile devices.
It's also studying customer comfort levels with automated payments. At this point, it's not ready to get rid of cashiers — they'll just be part of a bigger menu of customer checkout choices, according to Walmart rep Ragan Dickens.
Dickens would not comment on whether fully automated stores are in its plans. But for the past two months, it's been testing an Apple-store type concept called "Check out with Me" at 350 stores. It gives the agent the ability to check out a customer on a mobile device. Once the transaction is completed, a copy of the digital receipt is sent to the customer via email or text. Dickens said early results are encouraging, and the retailer plans to roll out this service to more stores.
Walmart's experiment with combining the physical and digital retail experience is an example of how a large, legacy retailer is combining the strength of a long-established physical presence combined with tech innovations — a hybrid model that may fit the needs of a broad swath of customers with varying levels of comfort with technology. Its tests to reduce dependence on traditional checkout counters center around two models: scan and go, which requires the customer to download an app or use a store-provided mobile device to self-scan items, and an Apple store-type model which puts the cashier capability in the hands of roving employees, who, with mobile devices in hand, complete checkouts for customers.
Big-box retailers are hot on the trail of a physical shopping experience that doesn't involve traditional cashiers. The front-runner, of course, is Amazon, which is planning the launch of a a second Amazon Go cashier-less store in Seattle this fall, and plans to open similar stores in Chicago and San Francisco. Walmart has been on the checkout automation journey since 2013, when it launched the first iteration of app-based self scan capabilities. The company said its vision is a physical retail environment that caters to different types of customers — from those who want an automated experience to those who feel they need to talk to a staff member.
After two attempts at scan and go, Walmart ditched the idea altogether, citing low adoption and a lack of customer comfort with the process. Walmart's first trial, rolled out in 2013 initially with 70 stores on board, involved using an app to scan items; a QR code generated at the conclusion of the shopping trip would be scanned at a self checkout counter before the customer paid. It rolled out a second scan-and-go pilot at 120 stores in August 2017, which expanded to an additional 120 stores in January of this year. It let the customer carry out in-app payments, eliminating the need for the checkout counter altogether. As an additional check, customers had to show an agent proof of purchases prior to leaving the store — a step many weren't comfortable making, the company said. The retailer wrapped up the second phase of the pilot in April.
"The friction point was that customers weren't used to the process of having a cart full of unbagged items, and having their receipts checked at the door," Dickens said.
Scan and go, however, is still available at Sam's Club locations, where customers are accustomed to having their receipts checked by an agent prior to departure, according to Dickens. As for Walmart's retail stores, the retailer isn't in a rush to re-introduce it. For now, it said it's using what it's learned from the pilot to improve it, and it's considering other services in the future, without specifying which ones.
Other retailers are also testing scan and go-type technology. A form of scan and go ("Scan, Bag, Go") is still being used at 400 Kroger stores, a feature it unveiled in January, and U.K. retailer Tesco is reportedly testing a similar feature. Meanwhile, Target and Home Depot are investing in making self-checkout machines work more efficiently; Home Depot is installing lockers at physical locations for customers to pick up prepaid online purchases.
Self-scanning is an imperfect solution because it takes time and may seem like a lot of work for a customer who seeks a digital-only, quick shopping experience, said Andrew Murphy, managing partner at Loup Ventures, a Minneapolis-based venture capital firm that invests in retail technology startups.
"Scan and go is a stepping stone to a fully automated checkout," he said. "Part of the clunkiness about it is that you still need to manually scan each item — in the future, your basket will already know what's in it," he said.
Amazon's "just walk out" technology lets customers open and scan their Amazon app as they enter the store, pick up the items they want and the customer is charged automatically. But despite the appeal of quick 'walk in, walk out' checkouts for consumers, the benefit to retailers is unclear, especially since (usually) the easier the transaction, the smaller the transaction, said Forrester principal analyst Sucharita Kodali. Ease of payment may prompt more frequent visits, but evidence around that is still anecdotal, she added. On top of that, Amazon Go-type technology it comes with a stiff price tag.
"The cost of the product has to be commensurate with the value it provides both the retailer and the customer — for one store it would probably be several million dollars' capital expense," said Kodali.
To make the technology scalable, the price would need to come down to around $100,000 per store, she added, and challenges like the necessity for an app and ensuring robust theft prevention measures would need to be worked out.
Plus, it remains unproven. "[The technology] is interesting, it's really expensive and cumbersome and hasn't even proven to work beyond 2000 square feet of an Amazon store," said Kodali.


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'Slash Film: Treasure Planet's Animation Was Even More 3D Than It Seems'
Let's get the obvious out of the way first: A drawing is two dimensional. It's got an X axis and a Y axis but, much like the sixth generation of "Pokémon," there's no Z in sight. Functionally, that means that every frame of a 2D animation project is planned ahead of time so that the renders will all match up. It also means that the camera angle is factored into each hand-drawn frame from the very beginning. In a 2002 interview with 70mm, Roy Conli — a producer on "Treasure Planet" — shared how that just wasn't enough freedom for John Musker or Ron Clement:
"We were able to employ a technology that was developed during 'Tarzan' called Deep Canvas, which allowed us to build sets in the computer, three-dimensionally. It's almost as if you're going in on a live-action set, and stroke-for-stroke, painting it as if you're painting a regular background. Only what you're utilizing is a digital paint brush that's being controlled by a stylist and a digital paint palette."
Conli went on to explain that, unlike "Tarzan," where the digital set painting was done on a "shot based" system where detail was only added where it was needed, the "Treasure Planet" sets were painted in their entirety, from every angle, so that camera experimentation could occur during production. Now think back to how many CGI sets were featured in "Treasure Planet." Montressor. Crescentia. Literal space, brimming with space whales (oh, that's where James Cameron got them from). Treasure freaking Planet itself? All CGI. Does that mean that a ton of effort went unseen? Absolutely. Is it still fascinating to think about? 100 percent yes.
/Film – 'Slash Film: Treasure Planet's Animation Was Even More 3D Than It Seems'
Author: Cameron Roy Hall
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John McWhorter Quotes
Anthony J. D'Angelo
William Arthur Ward
Find John McWhorter on:
American - Writer
Every third person in the world is a drama queen. And crying 'victim,' especially when you're not really a victim in any real way, feels good. It feels good to cry victim if you're not one.
John McWhorter
Good You Queen World
Texting is fingered speech. Now we can write the way we talk.
Way Speech Now Talk
It would be good if teachers could genuinely understand that black English is not mistakes, it's just different English, and that what you want to do is add an additional dialect to black students' repertoire rather than teaching them out of what's thought of as a bad habit, like sloppy posture or chewing with your mouth open.
Good You Black Mistakes
'LOL' is one of several texting expressions that convey nuance in a system where you don't have the voice and face to do it the way you normally would.
You Face Way Voice
People have been warning us that language was going to the dogs ever since Latin started turning into French. Yet the dogs in question never seem to emerge yelping on the horizon.
People Horizon Never Language
Ebonics - or black English, as I prefer to call it - is one of a great many dialects of English. And so English comes in a great many varieties, and black English is one of them.
Black Great English Many
Texting is very loose in its structure. No one thinks about capital letters or punctuation when one texts, but then again, do you think about those things when you talk?
You Think Things Talk
Loving your language means a command of its vocabulary beyond the level of the everyday.
Communication Your Loving Language
As languages go, English is pretty user friendly. If you look at a tiny language spoken somewhere that most of us have never heard of, chances are it's going to be so complicated that you have a hard time imagining how people can walk around speaking it without having a stroke.
Time You Walk People
Racism is not dead. Definitely, there are these biases.
Racism Dead Definitely Biases
As far as I'm concerned, and this is a big theme of mine, I'm not interested in white people loving me. It's an unrealistic expectation. Black people don't love anybody but themselves.
Love Me People Black
The only way that residual racist feelings could affect legislation, in my opinion, is through a lack of priorities, from not doing things.
Way Feelings Opinion Priorities
Rap and spoken word have reawakened the country to poetry in itself. Texting and Twitter encourage creative uses of casual language, in ways I have celebrated widely. But we've fallen behind on savoring the formal layer of our language.
Poetry Creative Country Language
The contribution of West African languages to Ebonics is absolutely infinitesimal. What it actually is is a very interesting hybrid of regional dialects of Great Britain that slaves in America were exposed to because they often worked alongside the indentured servants who spoke those dialects that we often learn about in school.
America School Great Contribution
Black English is something which - it's a natural system in itself. And even though it is a dialect of English, it can be very difficult for people who don't speak it, or who haven't been raised in it, to understand when it's running by quickly, spoken in particular by young men colloquially to each other. So that really is an issue.
People Black Men Understand
People banging away on their smartphones are fluently using a code separate from the one they use in actual writing, but a code it is, to which linguists are currently devoting articles.
People Writing Use Away
Most languages spoken by a few thousand people are so complicated they make your head swim; a Siberian yak herder's language is much more complicated than a Manhattan bond trader's.
People Bond Your Language
Black English is simpler than standard English in some ways; for example, it often gets by with just 'be' and drops 'am,' 'is,' and 'are.' That's because black English arose when adult African slaves learned the language.
Black Drops Language Just
People think of black English as ungrammatical, but it bears the same relationship to standard English as contemporary Hebrew does to ancient Hebrew.
Relationship People Black Think
As a linguist, I see the arbitrariness of strictures editors force on me as a writer.
Me See Force I See