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The Story of the Strathmore Municipal Library
Oct. 02, 2019 Heather Hobma
What has your library done for you?
This may seem like a strange question, because libraries obviously lend out books. And, if you're reading this, you likely spend at least some time here, or at least have borrowed a book or two. But have you ever really thought about what your library does? Maybe you have a personal goal to read every James Patterson novel and your library gives you that opportunity. Or maybe you like the access to the free public computers. Or maybe you hold a grudge against your junior high librarian for making you learn about the Dewey Decimal System and have vowed to never enter another library again. Because really, what can a library really do for you?
Some time between 1945 and 1948, Samuel Crowther believed that a library was a necessary addition to Strathmore. At the time, Strathmore didn't have a library. In fact, it was a 40-year project of love by a few very (very) determined individuals just to get it started. The library you see now is a modern conception, and even more so since the recent renovations. Crowther started the committee in the late '40s, which began by collecting used books for the catalogue. They were given matched funds from the Alberta Government up to $330 a year and a $25 donation from the Town of Strathmore. Even with governmental help, the committee relied mostly on funds raised during the annual bake sales and raffles and volunteer power, which we still heavily rely on today.
In 1967, the Town of Strathmore established the library as a municipal body, which guaranteed greatly needed per capita funding. During the 1970s, the library started collaborating more closely with the provincial government, which started funding a training program for small libraries in 1977.
During its tenure, the Library struggled to find a home and ended up moving several times: its first place was in the old town hall, but was soon moved to the Samuel Crowther School, and then into the new town office. In 1977, it moved into a shack by the Strathmore Hotel, and then moved to a slightly larger building in 1978, and then to a building on the main street in 1980. By 1982, the library board finally raised enough to move the library into a new facility, and the library/community centre, which opened its doors in 1983.
In its new home, the Library became an established institution for the Town, but what does it really do for the people who live and visit here?
Do you know that the Library brings in the latest books, magazines, movies, and music? And providing unique and tailored programs and special events, most of which that are free to attend?
Did you know, with your library card, you have access to our full catalogue and those of other libraries across Alberta? If we don't have what you need, we can bring it in for you! When you register with Me Libraries, you can borrow from the Calgary Public Library or Edmonton Public Library, and the Alberta Library gives you access to post-secondary and special libraries.
Maybe you're not really into books. Not everyone is. But do you listen to music or are you learning a foreign language or do you want to share an interactive story with your kids? With your library card, you have access to our e-resources, like Hoopla, NoveList Plus and NoveList Plus K-8, OverDrive/Libby, Pronunciator Language Learning, Solaro, TumbleBooks, and so many more. Our own patrons also have access to our eclectic Smore Stuff Collection, which includes life vests, STEAM kits, various tools, games, activities, and Audio-Visual equipment.
Maybe you're thinking that the library can't do much for you because you don't have a library card. Don't worry; we're not an exclusive club. You have access to a number of our services even without a library card. You can rent our rooms, or write an invigilated exam, or study in a quiet space, receive technology tutoring, or use our free WiFi and public computers. You can print, fax, or laminate here as well, and then enjoy our monthly art exhibitions that highlight a local artist. And beyond our walls, you can borrow books from the Little Free Libraries posted throughout the community or join our Visiting Library program if you can't leave your house.
We host a number of special events and programs throughout the year that are free and open to all community members, no library card needed, such as Nighttime Tales, Stitch Together, Baby Goose Rhyme Time, L'il Explorers, Crafternoon, and Budding Bookworm. Our events include book sales, Friday Fun Films, the Halloween Costume exchange, the Christmas Market, among many others.
But maybe you don't feel your library can do much for you. Sometimes when we ask this question, we hear back. Below are just a few things our library has done for a few of our community members and visitors:
"Our family is on a very tight budget, so we went to movie night at the Strathmore Library. It's free and you can bring your own snacks!"
"I am a mother of a preschool child. I spend most of my time tending to the needs of my child. Recently I attended a Storytime at the Strathmore Public Library and met other moms that I can share experiences with. I have made new friends and my child is learning to read books."
"I unexpectedly lost my job after being in the same position for over fifteen years. I did not have a computer or access to the Internet at home. The Strathmore Municipal Library provided the ability to access required government forms to fill out Employment insurance and other documentation through the public access computers. I took advantage of many resources including the e-Resource Career Cruising to help me create a resume and search for jobs and apply for them online."
"I do not live in Strathmore, but I was visiting my friend who lives here. I went to the Library and was able to use the internet to connect with my family while I was away. Being able to connect with them gave me peace of mind and I did not feel homesick."
We continue to grow and expand our programs and services. Your library is and will always be the hub of access, to resources, to knowledge, to entertainment, and to literacy and education. Our mission is to provide opportunities for life-long learning to the residents and visitors of Strathmore and District. Like those before us, our staff and volunteers are dedicated to supporting the freedom to read and access to lifelong learning, to providing cultural enrichment and education, and to helping improve the lives of our friends and neighbours and help them learn and grow. There is something for everyone off all ages at the Strathmore Municipal Library.
So, tell us, what has your library done for you?
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Protons are released in pain-generating pathological conditions such as inflammation, ischemic stroke, infection, and cancer. During normal synaptic activities, protons are thought to play a role in neurotransmission processes. Acid-sensing ion channels (ASICs) are typical proton sensors in the central nervous system (CNS) and the peripheral nervous system (PNS). In addition to ASICs, capsaicin- and heat-activated transient receptor potential vanilloid 1 (TRPV1) channels can also mediate proton-mediated pain signaling. In spite of their importance in perception of pH fluctuations, the regulatory mechanisms of these proton-sensitive ion channels still need to be further investigated. Here, we compared regulation of ASICs and TRPV1 by membrane phosphoinositides, which are general cofactors of many receptors and ion channels. We observed that ASICs do not require membrane phosphatidylinositol 4-phosphate (PI(4)P) or phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) for their function. However, TRPV1 currents were inhibited by simultaneous breakdown of PI(4)P and PI(4,5)P2. By using a novel chimeric protein, CF-PTEN, that can specifically dephosphorylate at the D3 position of phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3), we also observed that neither ASICs nor TRPV1 activities were altered by depletion of PI(3,4,5)P3 in intact cells. Finally, we compared the effects of arachidonic acid (AA) on two proton-sensitive ion channels. We observed that AA potentiates the currents of both ASICs and TRPV1, but that they have different recovery aspects. In conclusion, ASICs and TRPV1 have different sensitivities toward membrane phospholipids, such as PI(4)P, PI(4,5)P2, and AA, although they have common roles as proton sensors. Further investigation about the complementary roles and respective contributions of ASICs and TRPV1 in proton-mediated signaling is necessary.
Funding: This work was supported by the Ministry of Education, Science & Technology (No. 2012R1A1A2044699) and the DGIST R&D Program of the Ministry of Science, ICT&Future Planning (No. 14-BD-06). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Although the physiological pH level in the body is controlled by homeostatic mechanisms, transient and localized pH changes can occur in pain-generating pathological conditions and during normal synaptic activities [1,2]. Tissue acidosis is triggered during pain-producing inflammation, infection, ischemic stroke, and tumor development [1,3]. Severe ischemic stroke can reduce the extracellular pH level to 6.3 or even lower . Since local pH changes occur during normal brain activities , detection of pH fluctuations in the brain is also important. The proton has been recently reported as a neurotransmitter that can induce excitatory postsynaptic currents and regulate synaptic plasticity in the lateral amygdala . For that reason, it is not surprising that intrinsic systems capable of sensing pH fluctuations exist in our bodies.
Acid-sensing ion channels (ASICs), which belong to the epithelial Na+ channel (ENaC)/degenerin (DEG) superfamily of ion channels, are responsible for perception of pH changes in the central nervous system (CNS) and the peripheral nervous system (PNS) . These channels are voltage-independent, proton-gated cation channels mainly permeable to Na+ ion. Until recently, it has been reported that four genes, ACCN2, ACCN1, ACCN3, and ACCN4, encode at least six subunits, ASIC1a, ASIC1b, ASIC2a, ASIC2b, ASIC3, and ASIC4 [1,6]. Each subunit consists of two transmembrane (TM) domains, a large extracellular loop, and short intracellular N- and C-termini. They can form homo- or hetero-trimeric channels, while ASIC2b and ASIC4 only contribute to the functional properties of heteromeric channels containing each of them rather than the formation of homomeric channels. Capsaicin- or thermal stimuli-activated transient receptor potential vanilloid 1 (TRPV1) channels are also proton sensors primarily expressed in sensory neurons [7–9]. These polymodal signal integrators have received attention as therapeutic targets for pain. Four subunits are required to form a functional TRPV1 channel. Each subunit is composed of six TM domains, a pore domain between TM5 and TM6, and intracellular N- and C-termini . When opened by thermal or chemical noxious stimuli, these channels are permeable to nonselective cations with high Ca2+ permeability. Ca2+ influx through TRPV1 induces robust depletion of phosphatidylinositol 4-phosphate (PI(4)P) and phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) via the activation of phospholipase C-δ (PLCδ) isoforms and leads to desensitization of channels [10,11]. TRPV1 has putative proton binding sites on the extracellular face of the channel protein [7,9] as ASICs . TRPV1-mediated proton sensing is physiologically relevant to perception of nociceptive and inflammatory pain signaling in primary afferent neurons .
A signaling lipid, PI(4,5)P2, which is a minor acidic phospholipid in the inner leaflet of the eukaryotic cellular membranes, has received attention as a functional cofactor for membrane receptors and ion channels [14,15]. Cleavage of PI(4,5)P2 by receptor-activated PLC generates two second messengers: membrane-bound lipid diacylglycerol (DAG) and soluble inositol 1,4,5-trisphosphate (IP3). Depletion of PI(4,5)P2 during PLC signaling also inhibits the currents of several ion channels , including inwardly rectifying K+ (Kir) channel , KCNQ channel [17,18], voltage-gated Ca2+ channel (VGCC) , ENaC [20–22], and several members of the TRP channel family .
Although many interesting studies revealed that the plasma membrane (PM) phosphoinositide PI(4,5)P2 is a regulator of TRPV1 channels, whether PI(4,5)P2 has inhibitory or potentiating effects on their activities has been debated over the past decade [10,11,24–30]. In the case of ASICs, whether these channels have dependence on the phosphoinositides for their function has not been determined yet . Dorofeeva et al. (2009)  reported that homomeric ASIC1a channels can be inhibited by the activation of Gq-coupled M1 muscarinic receptor (M1R), which leads to hydrolysis of both PI(4)P and PI(4,5)P2 through the activation of PLCβ enzymes [32,33]. However, Li et al. (2012)  were not able to observe the decrease of ASIC1a currents during muscarinic receptor activation. Therefore, it is necessary to further examine the dependence of ASICs on membrane phospholipids for their function.
In this study, we focused on determining the sensitivities of ASICs toward phospholipids by comparing them to proton-sensitive TRPV1 channels. We used the recently developed translocatable pseudojanin (PJ) system  for investigating the sensitivities of ASICs to PM PI(4)P and PI(4,5)P2. In addition, we generated a novel inducible 3-phosphatase that can specifically dephosphorylate PM phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3) and investigated PI(3,4,5)P3 sensitivities of ASICs and TRPV1 channels in intact cells. Our studies demonstrate that the sensitivities of proton-sensitive ion channels toward PM phospholipids differ significantly depending on the type of channels.
Mouse cDNA clones of ASIC1a, ASIC2a, and ASIC3 were generously given to us by Michael J. Welsh (University of Iowa, Iowa city, Iowa). For the C-terminal fusion of GFP to each ASIC subunit, the cDNAs encoding ASIC1a, ASIC2a, and ASIC3 were amplified by the PCR using the primers (forward primer, 5'-GAATTCATGGAACTGAAGACCGAG-3', reverse primer, 5'-GGATCCCGGCAGGTAAAGTCCTCAAA-3', for ASIC1a-GFP; forward primer, 5'-GAATTCATGGACCTCAAGGAGAGC-3', reverse primer, 5'-GGATCCCGGCAGGCAATCTCCTCCAG-3', for ASIC2a-GFP; and forward primer, 5'-GAATTCATGAAACCTCCCTCAGGA-3', reverse primer, 5'-GGTACCGTGAGCCTTGTCACGAGGTA-3', for ASIC3-GFP), and inserted into the pEGFP-N1 (Clontech) vector. For the generation of CF-PTEN chimera, the DNA fragment encoding codons 22–403 was amplified by the PCR from Ci-VSPTEN21 , a kind gift from Carlos A. Villalba-Galea (Virginia Commonwealth University, Richmond, Virginia), using forward primer, 5'-AAGCTTCGGACTTAGACTTGACCTATA-3', reverse primer, 5'-GGATCCGACTTTTGTAATTTGTGAA-3', and fused to the C-terminus of CFP-FKBP. The following plasmids were generously given to us: Pseudojanin (PJ), PJ-Dead, PJ-Sac, INPP5E, LDR, PLCδ1-PH-GFP (from Bertil Hille, University of Washington School of Medicine, Seattle, Washington); Osh1-PH-GFP (from Deok-Jin Jang, Kyungpook National University, Sangju, Korea); rat TRPV1 with internal ribosome entry site EGFP and rat TRPV1 without GFP (from Jae-Yong Park, Korea University, Seoul, Korea); and Btk-PH-GFP (from Carlos A. Villalba-Galea, Virginia Commonwealth University, Richmond, Virginia).
TsA201 cells, derived from human embryonic kidney 293 cells (293tsA1609neo) by stably transfecting with the SV40 T-antigen  were obtained from Bertil Hille (University of Washington School of Medicine, Seattle, Washington). The cells were cultured in DMEM supplemented with 10% FBS and 0.2% penicillin/streptomycin at 37°C with 5% CO2 and transiently transfected using Lipofectamine 2000 (Invitrogen) with various cDNAs. For homomeric ASICs expression, cells were transfected with cDNA encoding ASIC1a-GFP, or ASIC2a-GFP, or ASIC3-GFP. For heteromeric ASICs expression, cells were transfected with different ASIC subunits in a 1:1 molar ratio, and 0.2 μg of cDNA encoding GFP was co-transfected as a marker for successfully transfected cells. GFP-positive cells were selected for recording the ASIC currents. For TRPV1 expression, cells were transfected with cDNA encoding TRPV1 with or without GFP. When needed, 0.2 μg of cDNA encoding tetrameric red fluorescence protein (DsRed) was co-transfected with TRPV1 as a marker for successfully transfected cells. The next day, cells were plated onto poly-L-lysine (0.1 mg/ml, Sigma) coated chips, and the fluorescent cells were studied within 2 days after transfection. Rapamycin (LC Laboratories), arachidonic acid (Sigma), amiloride (Tocris), and capsazepine (Sigma) were dissolved in DMSO (Sigma) to make stock solutions. Stock solutions were diluted in Ringer's solution before use.
The whole-cell configuration of the patch clamp technique was used to voltage-clamp at room temperature (22–25°C). Electrodes pulled from glass micropipette capillaries (Sutter Instrument) had resistances of 2–2.5 MΩ, and series resistance errors were compensated by > 60%. Fast and slow capacitances were compensated before the application of test-pulse. Recordings were performed using a HEKA EPC-10 amplifier with pulse software (HEKA Elektronik). The external Ringer's solution used for recording ASIC currents contained 160 mM NaCl, 5 mM KCl, 1 mM MgCl2, 2 mM CaCl2, and 10 mM HEPES, adjusted to pH 7.4 with tetramethylammonium hydroxide. For acidic solutions below pH 6.0, HEPES was replaced with MES. The pipette solution contained 140 mM KCl, 5 mM MgCl2, 10 mM HEPES, 0.1 mM 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), 3 mM Na2ATP, and 0.1 mM Na3GTP, adjusted to pH 7.4 with KOH. ASIC currents were recorded by holding the cell at -70 mV. The external Ringer's solution used for recording TRPV1 currents contained 150 mM NaCl, 5 mM KCl, 1 mM MgCl2, 2 mM EGTA, 10 mM Glucose, and 10 mM HEPES, adjusted to pH 7.4 with NaOH. The pipette solution contained 135 mM CsCl, 5 mM MgCl2, 10 mM HEPES, 5 mM EGTA, 5 mM Na2ATP, and 10 mM Glucose, adjusted to pH 7.4 with CsOH. TRPV1 currents were recorded by holding the cell at -80 mV. The following reagents were obtained: BAPTA, Na2ATP, Na3GTP, EGTA, CsOH and tetramethylammonium hydroxide (Sigma), HEPES (Calbiochem), MES (Alfa Aesar), and other chemicals (Merck).
TsA201 cells were imaged 1–2 days after transfection on poly-L-lysine coated chips with a Carl Zeiss LSM 700 confocal microscope (Carl Zeiss AG) at room temperature. The external Ringer's solution contained 160 mM NaCl, 2.5 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 10 mM HEPES, and 8 mM Glucose, adjusted to pH 7.4 with NaOH. For time courses, cell images were scanned with a 40 X (water) objective lens at 512 X 512 pixels using digital zoom. During time course experiments, images were taken every 5 s. Quantitative analysis of the cytosolic fluorescence intensity was performed using the 'measure' tool for the region of interest in ZEN 2012 lite imaging software (Carl Zeiss MicroImaging). All confocal images were transferred from LSM5 to JPEG format, and raw data from time courses was processed with Microsoft Office Excel 2012 (Microsoft) and Igor Pro (WaveMetrics, Inc.).
Data acquisition and analysis used Pulse/Pulse Fit software in combination with an EPC-10 patch clamp amplifier (HEKA Elektronik). Further data processing was performed with Microsoft Office Excel 2012 (Microsoft) and Igor Pro (WaveMetrics, Inc.). All quantitative data are expressed as the mean ± SEM. Comparisons between two groups were analyzed using Student's t-test. Comparisons among more than two groups were analyzed using one-way ANOVA followed by Bonferroni post-hoc test. Comparisons among more than two groups with two independent variables were analyzed using two-way ANOVA followed by Bonferroni post-hoc test. Differences were considered significant at the * P < 0.05, ** P < 0.01, and *** P < 0.001 levels, as appropriate.
To assess the requirement of PM phosphoinositides for the activities of proton-sensitive ion channels, we employed the chemically-inducible dimerization (CID) system. Rapamycin-induced dimerization of FKBP (FK506 binding protein) and FRB (FKBP-rapamycin binding domain of mTOR) can be used to rapidly and irreversibly recruit the enzyme of interest to the target region inside the cell . We applied the recently described fusion protein mRFP-FKBP-Pseudojanin (PJ), which contains both inositol polyphosphate-5-phosphatase E (INPP5E) and sac1 phosphatase, and thus, the recruitment of which to the PM leads to simultaneous depletion of PI(4,5)P2 and PI(4)P . First, we tested the activity of the PJ construct by using the pleckstrin homology (PH) domain of PLCδ1 (PLCδ1-PH-GFP) and oxysterol-binding protein homologues (Osh1-PH-GFP) as indicators for PM PI(4,5)P2 and PI(4)P, respectively. PJ was localized to the cytoplasm when expressed in tsA201 cells. Recruiting PJ to the PM anchor LDR (N-terminal myristoylation and palmitoylation modification sequence of a Lyn kinase coupled to FRB domain) by the addition of 1 μM of rapamycin (Rapa) for 60 s resulted in the release of both Osh1-PH and PLCδ-PH to the cytoplasm in each separate confocal experiment (Fig. 1A, bottom). The cytosolic fluorescence intensity of PJ rapidly declined upon rapamycin addition, while that of Osh1-PH or PLCδ-PH increased (Fig. 1B, bottom). We also confirmed that the recruitment of PJ-Sac (INPP5E domain is inactivated by mutation to specifically dephosphorylate PI(4)P ) resulted in the release of Osh1-PH from the PM and increase in the cytosolic fluorescence intensity of Osh1-PH (Fig. 1A and B, middle). Recruitment of INPP5E to the PM decreased the PI(4,5)P2 level, which was confirmed by the translocation of PI(4,5)P2 probe to the cytoplasm with the increase in the cytosolic intensity of PLCδ-PH (Fig. 1A and B, middle). However, recruiting the cytosolic PJ-Dead, a chimera with inactivated sac1 and INPP5E , to the PM had no effects on the localization of Osh1-PH or PLCδ-PH (Fig. 1A, top). Consequently, the cytosolic fluorescence intensities of Osh1-PH and PLCδ-PH remained unchanged following the addition of rapamycin (Fig. 1B, top).
Fig 1. The activity of TRPV1 is dependent on phosphoinositides.
(A) Confocal images of cells expressing PJ-Dead (top), PJ-Sac (middle), INPP5E (middle), or PJ (bottom) with LDR and respective biosensors for PI(4)P (Osh1-PH-GFP) or PI(4,5)P2 (PLCδ1-PH-GFP). Images before and after the addition of rapamycin (1 μM) for 60 s (Scale bar, 5 μm). (B) Cytosolic fluorescence intensities of RFP (red) and GFP (green) for the cells in (A). The values of the Y-axis use an arbitrary unit. Cells expressing PJ-Dead (top), PJ-Sac (middle), INPP5E (middle), or PJ (bottom) (n = 3, respectively). (C) TRPV1 currents triggered by prolonged extracellular pH drop to 5.0 for 150 s. Rapamycin (1 μM) was co-applied for 90 s during the acid stimuli. Amiloride (300 μM) was pretreated for 30 s before the pH pulse. Black dashed line indicates the zero current level. Red dashed line indicates the point of rapamycin application. (D) Percentage of current decrease in (C) during 45 s of acidification before (grey) and after (red) rapamycin addition (n = 12 for PJ-Dead; n = 14 for PJ-Sac; n = 12 for INPP5E; n = 10 for PJ). ** P < 0.01 and *** P < 0.001, with two-way ANOVA followed by Bonferroni post-hoc test and one-way ANOVA followed by Bonferroni post-hoc test. Data are mean ± SEM.
We applied this system on acid-evoked TRPV1 currents to test the regulatory aspects of phosphoinositides reported by previous studies [10,11,24–30]. Since tsA201 cells have been reported to have endogenous ASIC currents , the cells transiently expressing TRPV1 and respective PJ system constructs were preincubated by amiloride (AMI), a general inhibitor of ASICs, before the acid stimulation to selectively measure the TRPV1 currents. After the preincubation of cells with extracellular solution containing 300 μM of amiloride for 30 s, the cells were stimulated by pH 5.0 solution for 60 s; then, 1 μM of rapamycin was co-applied with pH 5.0 solution for 90 s. Since we observed variable desensitization among the cells during prolonged acidification before the addition of rapamycin, we first compared how much the currents decrease during acidification in the absence of rapamycin among four groups (PJ-Dead, PJ-Sac, INPP5E, and PJ). TRPV1 currents usually reached the maximum current amplitude within 15 s after the currents were activated; thus, we compared the current decrease during 45 s of acidification after reaching the maximum current amplitude. The currents of TRPV1 in cells expressing PJ-Dead or PJ-Sac were desensitized by 7 ± 3% (n = 12) or 9 ± 3% (n = 14), respectively (Fig. 1C-D). When the cells were transfected with INPP5E or PJ, the currents were decreased by 14 ± 1% (n = 12) or 13 ± 6% (n = 10), respectively (Fig. 1C-D). Desensitization rates of currents showed no statistically significant differences among the four groups before the application of rapamycin. The currents in cells expressing PJ-Dead, PJ-Sac, and INPP5E were decreased by 16 ± 4% (n = 12), 23 ± 4% (n = 14), and 26 ± 4% (n = 12), respectively, during 45 s of acidification right after rapamycin addition (Fig. 1C-D). However, translocation of PJ to the PM by the application of rapamycin decreased TRPV1 currents by 48 ± 6% (n = 10) during 45 s of acidification (Fig. 1C-D). These results suggest that the activities of TRPV1 channels are diminished by simultaneous breakdown of both PM PI(4)P and PI(4,5)P2, as previously reported [11,35].
Next, we examined whether ASICs also require phosphoinositides for their function as TRPV1 channels. ENaC belongs to the same superfamily of ion channels as ASICs and is known to be regulated by PM PI(4,5)P2 and PI(3,4,5)P3 [20–22]. That means, ASICs could have sensitivities toward phosphoinositides. To test this, we employed the PJ system and observed the activities of homomeric and heteromeric ASICs. The cells transiently expressing each GFP-tagged ASIC subunit (ASIC1a, or ASIC2a, or ASIC3) and respective PJ systems were repetitively activated by extracellular acidification from pH 7.4 to pH 6.0 (ASIC1a or ASIC3) or to pH 4.5 (ASIC2a) for 10 s with 2 min time intervals. As previously reported, GFP fusion to the C-terminus of ASIC subunit did not affect the electrophysiological properties of wild-type ASIC currents expressed in tsA201 cells . For recruiting the interested enzyme to the PM, 1 μM of rapamycin was perfused for 60 s before the second pH pulse. To minimize possible side effects of rapamycin, normal extracellular solution was perfused right after the application of rapamycin for 10 s just before the second pH pulse. First, we confirmed that, as a control, the recruitment of PJ-Dead to the PM anchor does not affect the repetitive proton-activated ASIC1a currents except for tachyphylaxis (reduction in current amplitude with repeated stimulation), a unique property of homomeric ASIC1a channels  (Fig. 2A-B). Then, we observed that translocation of PJ-Sac or INPP5E to the PM to specifically deplete PM PI(4)P or PI(4,5)P2, respectively, had no effects on the relative current density of homomeric ASIC1a channels (Fig. 2A-B). Simultaneous depletion of both PI(4)P and PI(4,5)P2 by PJ also had no significant effect on the successively triggered ASIC1a currents (Fig. 2A-B). We found that neither ASIC2a nor ASIC3 homomeric channels were affected by the recruitment of PJ to the PM (Fig. 2C-D), allowing us to conclude that unlike proton-sensitive TRPV1 channels, the activities of homomeric ASICs are independent of PM PI(4)P and PI(4,5)P2.
Fig 2. Homomeric ASIC currents are insensitive to PI(4)P and PI(4,5)P2.
(A) ASIC1a currents evoked by repetitive rapid extracellular pH change from 7.4 to 6.0 for 10 s with time intervals of 120 s in cells expressing LDR and either PJ-Dead, PJ-Sac, INPP5E, or PJ. Rapamycin (1 μM) was bath-applied for 60 s, and then normal extracellular solution was perfused for 10 s right before the second pulse to minimize possible side effects of rapamycin. Dashed line indicates the zero current level. (B) Relative current density measured for the cells in (A) (n = 6, respectively). Current density of each pulse was divided by that of the first pulse. There is no statistical significance with two-way ANOVA followed by Bonferroni post-hoc test. (C) ASIC2a or ASIC3 current traces evoked by pH drop to 4.5 or 6.0 for 10 s. (D) Relative current density measured for the cells in (C) (n = 6, respectively). There is no statistical significance with two-way ANOVA followed by Bonferroni post-hoc test. Data are mean ± SEM.
We also tested whether heteromeric ASICs have dependence on phosphoinositides for their function, since most ASICs exist as heteromeric channels in physiological conditions [42–44]. The current traces from heteromeric channels of ASIC1a/2a, ASIC1a/3, and ASIC2a/3 were similar to those of a previous study . Recruitment of PJ to the PM had no significant effects on either ASIC1a/2a or ASIC1a/3 heteromeric channels (Fig. 3A-B), and transient and sustained currents of ASIC2a/3 heteromeric channels were not significantly affected by the application of rapamycin (Fig. 3A and C). In conclusion, neither homomeric ASICs nor heteromeric ASICs require phosphoinositides for their activities.
Fig 3. Heteromeric ASIC currents are insensitive to PI(4)P and PI(4,5)P2.
(A) Current traces from ASIC1a/2a, ASIC1a/3, and ASIC2a/3 heteromeric channels evoked by extracellular acidification in cells expressing LDR and PJ. Rapamycin (1 μM) was bath-applied for 60 s, and then normal extracellular solution was perfused for 10 s right before the second pulse to minimize possible side effects of rapamycin. Dashed line indicates the zero current level. (B) Relative current density measured for the currents of ASIC1a/2a and ASIC1a/3 in (A) (n = 3, respectively). Current density of each pulse was divided by that of the first pulse. There is no statistical significance with two-way ANOVA followed by Bonferroni post-hoc test. (C) Relative current density measured for the transient and sustained currents of ASIC2a/3 in (A) (n = 3 for PJ-Dead; n = 5 for PJ). There is no statistical significance with two-way ANOVA followed by Bonferroni post-hoc test. Data are mean ± SEM.
Even though the PJ system is a powerful tool for probing the role of phosphoinositides for the function of ion channels, it has a limitation in terms of investigating the specific effect of PI(3,4,5)P3 on the channels. Therefore, we generated a novel chimeric protein to further investigate the role of PI(3,4,5)P3 in the activities of proton-sensitive ion channels. One of the tumor suppressor genes, PTEN (phosphatase and tensin homologue deleted on chromosome 10) codes a cytosolic 3-phosphatase that degrades PI(3,4,5)P3 by removing the phosphate at the D3 position of the inositol ring [46–48]. PTEN has a substrate specificity toward PI(3,4,5)P3  and is composed of N-terminal phosphoinositide-binding motif (PBM) that contributes to the recruitment of PTEN to the PM, phosphatase domain (PD), C2 domain (C2) that binds to PM phosphatidylserine (PS), and C-terminal tail PDZ-binding domain [36,46–48] (Fig. 4A). We inserted the region from PD to C-terminal tail of PTEN to the C-terminus of CFP-FKBP (CF) (Fig. 4A). CFP-FKBP-PTEN (CF-PTEN) was translocated to the PM anchor LDR upon addition of 1 μM rapamycin; in turn, PI(3,4,5)P3 was depleted as shown by the specific PI(3,4,5)P3 probe, PH domain of Bruton tyrosine kinase (Btk) (Btk-PH-GFP) (Fig. 4B). The cytosolic fluorescence intensity of Btk-PH rapidly increased while that of CF-PTEN gradually declined by rapamycin addition (Fig. 4B), indicating the successful development of a novel translocatable 3-phosphatase tool.
Fig 4. Neither ASICs nor TRPV1 activities are affected by depletion of PI(3,4,5)P3.
(A) CF-PTEN is rapidly recruited to the plasma membrane anchor LDR by dimerization of FRB and FKBP upon addition of rapamycin, and the PD domain of CF-PTEN specifically dephosphorylates PI(3,4,5)P3 to PI(4,5)P2. Red bar in the C-terminal tail of PTEN indicates PDZ-binding domain. (B) Confocal images of cells expressing LDR, CF-PTEN, and Btk-PH-GFP before and after the addition of rapamycin (1 μM) for 120 s (Scale bar, 5 μm) and cytosolic fluorescence intensities of CFP (blue) and GFP (green) (n = 3). The values of the Y-axis use an arbitrary unit. (C) ASIC current traces triggered by extracellular acidification in cells expressing LDR and CF (lacking PTEN) or CF-PTEN. Rapamycin (1 μM) was bath-applied for 60 s, and then normal extracellular solution was perfused for 10 s right before the second pulse to minimize possible side effects of rapamycin. Dashed line indicates the zero current level. (D) Relative current density measured for the cells in (C) (CF (n = 8), CF-PTEN (n = 7) for ASIC1a; CF (n = 8), CF-PTEN (n = 8) for ASIC2a; and CF (n = 10), CF-PTEN (n = 10) for ASIC3). Current density of each pulse was divided by that of the first pulse. There is no statistical significance with two-way ANOVA followed by Bonferroni post-hoc test. (E) TRPV1 currents in response to pH drop in the cells expressing LDR and CF or CF-PTEN. Rapamycin (1 μM) was co-applied for 90 s during the acid stimuli. Amiloride (300 μM) was pretreated for 30 s before the pH pulse. Black dashed line indicates the zero current level. Red dashed line indicates the point of rapamycin application. (F) Percentage of current decrease in (E) during 45 s of acidification before (grey) and after (red) rapamycin addition (n = 9 for CF; n = 10 for CF-PTEN). Data are mean ± SEM.
Using this tool, we tested the sensitivities of homomeric ASICs and TRPV1 channels to PM PI(3,4,5)P3. Although the membrane PI(3,4,5)P3 was depleted following the addition of rapamycin, the repetitively activated currents of homomeric ASIC1a, ASIC2a, and ASIC3 channels remained unchanged (Fig. 4C-D). These results suggest that ASICs are insensitive to the depletion of PI(3,4,5)P3 in intact cells. Next, we investigated whether the depletion of membrane PI(3,4,5)P3 in an intact cell affects the proton-activated TRPV1 currents. The current decrease during 45 s of acidification right before the addition of rapamycin was not significantly different between two groups (CF: 8 ± 2% (n = 9) and CF-PTEN: 10 ± 3% (n = 10)) (Fig. 4E-F). In the presence of rapamycin, TRPV1 currents were decreased by 18 ± 4% (n = 9) or 21 ± 4% (n = 10) in cells expressing CF or CF-PTEN, respectively, during 45 s of acidification (Fig. 4E-F). Difference in current decrease ratio between the two groups was not statistically significant. These results indicate that the depletion of PI(3,4,5)P3 has no effects on the activities of TRPV1 channels, although TRPV1 currents are inhibited by simultaneous depletion of both PI(4)P and PI(4,5)P2. This result is supported by the previous reports suggesting that PI(3,4,5)P3 is not an endogenous regulatory factor of TRPV1 currents [27,28]. In inside-out patches, the application of pH domain of the general receptor of phosphoinositides 1 (GRP1) which selectively binds to PI(3,4,5)P3 did not inhibit the TRPV1 currents, while the application of PLCδ1-PH produced the decrease of the currents . Moreover, in a reconstituted liposome, PI(3,4,5)P3 did not have any effect on the TRPV1 currents, unlike other phosphoinositides . Taken together, neither ASICs nor TRPV1 activities are altered by depletion of PI(3,4,5)P3.
In the data above, we investigated the regulatory effects of PM phosphoinositides on proton-sensitive ion channels, ASICs and TRPV1, and discovered a difference in their sensitivities toward phosphoinositides. We next asked how these proton-sensitive ion channels are regulated by arachidonic acid (AA), which is liberated from the membrane phospholipids by phospholipase A2 (PLA2) activity [50,51]. A pro-inflammatory mediator, AA is a polyunsaturated fatty acid acting as a lipid second messenger [1,52]. Several ion channels, including ASICs [53,54] and TRP channels [28,55], are known to be regulated by AA either by a direct or indirect action . Here, we investigated whether AA also differentially regulates the activities of ASICs and TRPV1 channels as do phosphoinositides.
When the extracellular solution containing 10 μM of AA was perfused for 20 s right before the second pH pulse, the peak current density of the second pulse in cells transiently expressing homomeric ASIC1a channels was increased by 81 ± 29% (n = 6) compared to that of the first pulse (Fig. 5A-B). On the other hand, the peak current density of the second pulse was slightly decreased in the control group (Fig. 5B). We observed that the potentiating effect of AA is reversible, and the peak current density was recovered to the initial level after washout of AA (Fig. 5A-B). In the cells expressing ASIC2a homomeric channels, the peak current density of the second pulse was reversibly increased by 103 ± 30% (n = 10) compared to that of the first pulse, whereas, in the control group, the difference in the current density between the first and the second pulses was negligible (Fig. 5A-B). Similarly, the peak current density of the second pulse in cells expressing ASIC3 homomeric channels was reversibly increased by 133 ± 33% (n = 12) compared to that of the first pulse (Fig. 5A-B). AA increased the respective ASIC currents in a dose-dependent manner (Fig. 5C). ASIC1a, ASIC2a, and ASIC3 homomeric channels displayed similar dose-dependent curves.
Fig 5. Potentiation of ASICs by AA.
(A) ASIC current traces activated by rapid extracellular pH changes. AA (10 μM) was bath-applied for 20 s before the second pulse. Dashed line indicates the zero current level. (B) Relative current density was measured for the cells expressing ASIC1a (n = 5 for DMSO; n = 6 for AA), ASIC2a (n = 5 for DMSO; n = 10 for AA), and ASIC3 (n = 5 for DMSO; n = 12 for AA). Current density of each pulse was divided by that of the first pulse. * P < 0.05 and ** P < 0.01, with two-way ANOVA followed by Bonferroni post-hoc test and student's t-test. (C) Dose-dependent relative current density of ASIC1a (blue) (n = 5–20), ASIC2a (green) (n = 5–25), and ASIC3 (red) (n = 5–23). (D) ASIC1a and ASIC3 currents were inhibited by preincubation of cells with pH 7.4 solution containing amiloride (300 μM) for 20 s before the second pulse. In the case of ASIC2a, 600 μM of amiloride was applied for 30 s before and during the second pulse. (E) Percentage of inhibition by amiloride in the absence (grey) or the presence (yellow) of AA (AMI (n = 7) and AA+AMI (n = 4) for ASIC1a; AMI (n = 4) and AA+AMI (n = 3) for ASIC2a; and AMI (n = 6) and AA+AMI (n = 6) for ASIC3). (F) The potentiating effect of AA (10 μM) on ASIC currents was inhibited by amiloride. Data are mean ± SEM.
Next, we also tested whether the potentiated currents by AA are attributable to other non-specific currents in tsA201 cells. First, we observed that respective ASIC currents were inhibited by amiloride. ASIC1a currents were inhibited by 95 ± 1% (n = 7) when the extracellular solution containing 300 μM of amiloride was perfused for 20 s right before the second pH pulse (Fig. 5D-E). AA (10 μM)-induced increased ASIC1a currents were also inhibited by 93 ± 2% (n = 4) by co-application of AA and amiloride before the second pH pulse (Fig. 5E-F). In the case of ASIC2a homomeric channels, the currents were inhibited by 65 ± 3% (n = 4) when amiloride (600 μM) was applied for 30 s before and during the second pH pulse (Fig. 5D-E). The potentiating effects of AA (10 μM) on ASIC2a currents were inhibited by 72 ± 11% (n = 3) by amiloride (Fig. 5E-F). ASIC3 currents were inhibited by 80 ± 2% (n = 6) by preincubation of cells with extracellular solution containing 300 μM of amiloride for 20 s before the second pH pulse (Fig. 5D-E), and the potentiating effects of AA (10 μM) were similarly inhibited by 85 ± 1% (n = 6) (Fig. 5E-F). Collectively, our results suggest that AA reversibly potentiates homomeric ASIC currents.
At this time, we investigated the effect of AA on proton-activated TRPV1 currents. The bath-application of AA (2 μM) for 20 s right before the second addition of amiloride increased the current density of the second pulse by 129 ± 21% (n = 9) compared to that of the first pulse, whereas the difference in the current density between the first and the second pulses were insignificant (Fig. 6A-B). Unlike ASIC currents, AA-induced potentiated TRPV1 currents were not fully recovered to the initial level (Fig. 6A-B). AA potentiated the TRPV1 currents in a dose-dependent manner (Fig. 6C). We also tested whether these potentiated currents are indeed attributable to TRPV1 channels. TRPV1 currents were inhibited by 78 ± 7% (n = 4) by bath-application of capsazepine (CPZ) (10 μM), a specific TRPV1 inhibitor for 20 s (Fig. 6D and F). Co-application of capsazepine and AA (2 μM) before the second amiloride treatment inhibited the potentiating effect of AA on TRPV1 currents by 89 ± 5% (n = 4) (Fig. 6E-F). Taken together, AA enhances the activities of TRPV1 channels, although the recovery aspect differs from that of ASICs (Figs. 5B and 6B).
Fig 6. Potentiation of TRPV1 by AA.
(A) TRPV1 current traces repetitively activated by extracellular pH drop to 5.5 for 30 s with time intervals of 300 s. Amiloride (300 μM) was pretreated for 10 s before the pH pulses. AA (2 μM) was applied for 20 s before the second amiloride treatment. Dashed line indicates the zero current level. (B) Relative current density was measured for the cells in (A) (n = 4 for DMSO; n = 9 for AA). Current density of each pulse was divided by that of the first pulse. ** P < 0.01, with two-way ANOVA followed by Bonferroni post-hoc test and student's t-test. (C) Dose-dependent relative current density of TRPV1 (n = 5–17). (D) TRPV1 currents were inhibited by preincubation of cells with pH 7.4 solution containing capsazepine (10 μM) for 20 s before the second amiloride treatment. (E) The potentiating effect of AA (2 μM) on TRPV1 currents was inhibited by capsazepine. (F) Percentage of inhibition by capsazepine in the absence (grey) or the presence (blue) of AA (n = 6 for CPZ; n = 6 for AA+CPZ). Data are mean ± SEM.
In this study, we observed that the activities of ASICs are independent of membrane phosphoinositides such as PI(4)P, PI(4,5)P2, and possibly PI(3,4,5)P3 by using the translocatable phosphatase system. This is the first report to show the sensitivities of ASICs toward phosphoinositides with direct manipulation of membrane phosphoinositides in intact cells. On the other hand, studies about TRPV1 sensitivity toward phosphoinositides have been undertaken by various research groups. In our system, we observed that proton-activated TRPV1 currents can be regulated by simultaneous manipulation of PI(4)P and PI(4,5)P2 level, not PI(3,4,5)P3 level, in intact cells.
Although both ASICs and TRPV1 channels can sense proton-mediated signaling, their expression and biophysical properties are different. ASICs are widely distributed throughout the CNS and the PNS, and they act as transducers of multiple sensory and synaptic signaling. TRPV1 is highly expressed in peripheral nerve endings of primary sensory neurons. Multimodal response of TRPV1 to various noxious stimuli elicits pain and leads to the development of hyperalgesia . According to previous studies, ASICs and TRPV1 are co-localized in a great part of subpopulation of DRG neurons, although there are differences between species [56,57]. In rat DRG neurons, ASIC1a and ASIC3 transcripts were detected in approximately 40–45% and 30% of the TRPV1-positive neurons, respectively . In these subsets of native sensory neurons, it is highly possible that the currents elicited by tissue acidosis are contributed from both ASICs and TRPV1 channels.
It is noteworthy that two channels are activated by different ranges of pH value. TRPV1 is activated by more severe acidification (pH0.5 activation of 5.4) than that required to activate most ASIC subunits [1,58]. ASIC1a and ASIC3 are sensitive to moderate pH drop (pH0.5 activation of 6.2–6.8 for ASIC1a and 6.2–6.7 for ASIC3), whereas ASIC2a requires more severe acidification for activation (pH0.5 activation of 3.8–5.0) . Interestingly, extracellular protons have a dual effect on TRPV1 currents. At low pH (< 6.0), protons themselves activate the channel at room temperature. However, protons potentiate the channel already opened by other stimuli (capsaicin or heat) by lowering the threshold for channel activation at higher pH levels (6–7.4) [7,9,58]. Therefore, ASICs are considered main mediators of pain induced by moderate tissue acidosis, while TRPV1 is thought to contribute to more severe acidosis-mediated pain perception, together with ASICs in peripheral sensory neurons . Hence, the complementary roles of these two proton-sensitive ion channels, ASICs and TRPV1, have great significance for perception of pH changes; thus, to understand the regulatory mechanisms of those channels is quite important.
Phosphoinositides have emerged as general regulators of ion channels, and PI(4,5)P2 is known to stabilize the open state of many ion channels [1,14,17–23]. However, regulation of TRPV1 by phosphoinositides is controversial [10,11,24–30]. In this study, we simply tested the effects of phosphoinositides on proton-activated TRPV1 currents by using a rapamycin-inducible PJ system, and compared the results with that of ASICs. We observed that proton-activated TRPV1 currents are significantly inhibited by the recruitment of PJ, while the translocation of INPP5E had no statistically significant effect on the currents (Fig. 1). These results are consistent with the study by Hammond et al. (2012) . They observed that capsaicin-activated TRPV1 currents in HEK293 cells are inhibited when both PI(4)P and PI(4,5)P2 are depleted by the translocation of PJ . In contrast to TRPV1 channels, the function of ASICs does not rely on PM phosphoinositides. By using the PJ system, we confirmed that homomeric ASIC1a, ASIC2a, and ASIC3 channels and heteromeric ASIC1a/2a, ASIC1a/3, and ASIC2a/3 channels are insensitive to PI(4)P and PI(4,5)P2. These results are, in fact, unexpected since one previous study reported that activation of M1R by its agonist, oxotremorine-M (Oxo-M), inhibited ASIC currents in Chinese hamster ovary (CHO) cells heterologously expressing ASIC1a and M1R, and also in isolated rat hippocampus CA1 and striatum interneurons . That study suggested that muscarinic inhibition of ASIC1a currents could be due to depletion of PI(4,5)P2 available to the channel ; however, we observed no dependence of ASICs on PI(4)P or PI(4,5)P2 (Figs. 2 and 3). Therefore, it is possible that inhibition of ASIC currents by M1R activation might occur through mechanisms other than direct action of PI(4,5)P2 hydrolysis. We also tried to test whether the activation of M1R by Oxo-M modulates the function of homomeric ASIC1a channels in either tsA201 or CHO cells. However, we observed no inhibition of ASIC1a currents by M1R activation in either type of cells (unpublished observations). In another previous study by Li et al. (2012), they found that supplementing the pipette solution with a short-chain PIP2 was not effective to decrease the desensitization of ASIC1a currents. Furthermore, they observed that the currents were not regulated by activation of muscarinic receptor . The discrepancy between the studies is not clear; however, in our system, using the translocatable PJ system, we verified that the activities of ASICs are independent from PM phosphoinositides.
We also investigated the dependence of ASICs and TRPV1 channels on PI(3,4,5)P3. ENaC and some families of TRP channels such as TRPM4 are known to have sensitivities toward PI(3,4,5)P3 as well as PI(4,5)P2 [20–22,59]. To selectively dephosphorylate PI(3,4,5)P3, we generated a novel engineered phosphatase tool from PTEN, which is a well characterized 3-phosphatase that prefers PI(3,4,5)P3 as a substrate . By using a chimeric protein CF-PTEN, we observed that neither ASICs nor TRPV1 currents were altered by depletion of membrane PI(3,4,5)P3 in intact cells.
Finally, we compared differential regulatory features of ASICs and TRPV1 by AA, a pro-inflammatory mediator released from phospholipids. In our experiments, AA induced significant potentiation of all ASIC1a, ASIC2a, ASIC3, and TRPV1 channels. TRPV1 currents were particularly more sensitive to AA than ASIC currents were. It is noteworthy that potentiated ASIC currents by AA were, remarkably, almost fully recovered to the initial level of the currents, while TRPV1 currents were partially recovered after washout of AA (Figs. 5 and 6). We propose that this is likely due to the difference in regulatory mechanisms of those channels by AA. The potentiating effect of AA on TRPV1 currents has been reported to result from the AA metabolites of lipoxygenase pathways, such as 12- and 15-hydroperoxyeicosatetraenoic acid (HPETE) [60,61]. However, a recent study suggested that both AA and its metabolites can produce marked sensitization of TRPV1 currents in the reconstituted liposome, allowing them to observe the direct effects of factors in the absence of other cellular enzymes . Therefore, AA seems to regulate the TRPV1 channels by both a direct and an indirect action through the metabolism pathways. On the other hand, ASICs are thought to be directly regulated by AA . One previous study reported that inhibition of lipoxygenase or cyclooxygenase pathway did not impair the potentiating effect of AA on ASIC currents . Therefore, ASICs and TRPV1 channels are differentially regulated by a pro-inflammatory mediator AA.
In this study, we observed that two proton-sensitive ion channels, ASICs and TRPV1, display differential regulatory features by membrane phosphoinositides and AA. Their different topology, distribution, and biophysical properties might establish different sensitivities toward phospholipids. Understanding the relationship between these two groups of channels, and their relative contributions in proton-mediated signaling, is quite important for comprehending the complementary roles of ASICs and TRPV1 in the nervous system.
We are grateful to Dr. Bertil Hille for his valuable discussions. We thank the many labs that provided the plasmids.
Conceived and designed the experiments: BCS. Performed the experiments: HJK SYY DIK. Analyzed the data: HJK SYY DIK. Contributed reagents/materials/analysis tools: HJK SYY DIK. Wrote the paper: HJK BCS.
15. Falkenburger BH, Jensen JB, Dickson EJ, Suh BC, Hille B. Phosphoinositides: lipid regulators of membrane proteins. J Physiol. 2010;588(Pt 17): 3179–85.
23. Hardie RC. TRP channels and lipids: from Drosophila to mammalian physiology. J Physiol. 2007;578(Pt 1): 9–24.


text:
package org.higherstate.jameson.extractors

import org.higherstate.jameson.tokenizers.{BooleanToken, Tokenizer}
import org.higherstate.jameson.Path
import org.higherstate.jameson.failures._

trait BooleanExtractor[T] extends Extractor[Boolean,  T] {
  def parse(tokenizer:Tokenizer, path: Path) = tokenizer.head match {
    case BooleanToken(value) =>
      apply(value, path)
    case token              =>
      Failure(InvalidTokenFailure(this, "Expected Boolean token", token, path))
  }
}

redpajama_set_name: RedPajamaGithub


text:
The Psychology of Romantic Love
Romantic Love in an Anti-Romantic Age
By Nathaniel Branden
Category: Psychology | Personal Growth
About The Psychology of Romantic Love
What love is, why love is born, why it sometimes grows, and why it sometimes dies.
Have you ever wondered how romantic love evolves? What the difference is between mature and immature love? What role sex plays in romantic love, and whether love necessarily implies sexual exclusivity? And, most important, how can we make love last? Originally published in 1980, this updated edition of The Psychology of Romantic Love explores the nature of romantic love on many levels-the philosophical, the historical, the sociological, and the physiological. Nathaniel Branden explains why so many people say that romantic love is just not possible in today's world and-drawing on his experience with thousands of couples-finds that such love is still a possibility for anyone who understands its essence and is willing to accept its challenges.
Branden sees it as a pathway not only to extraordinary joy but also to profound self-discovery. His vision of love is thoroughly appropriate to our time and grounded in our humanness.
Also by Nathaniel Branden
See all books by Nathaniel Branden
About Nathaniel Branden
Nathaniel Branden's pioneering work in the field of self-esteem over seven decades contributed to the evolution of the concept from obscurity to greater levels of clarity and acceptance. He wrote nearly 20 books on self-esteem, including such influential works as The Psychology… More about Nathaniel Branden
Published by TarcherPerigee
Jan 31, 2008 | 240 Pages | 5-1/2 x 8-1/4 | ISBN 9781585426256
Jan 31, 2008 | 240 Pages | ISBN 9781101216149
People Who Read The Psychology of Romantic Love Also Read

redpajama_set_name: RedPajamaCommonCrawl


While Pranayama Standing Deep Breathing isn't strictly considered a posture it is an integral component to the Hot Bikram Yoga series. Don't be fooled that it is 'just' a breathing exercise, it is as important as every posture that follows.
correct alignment is key to everything we do in the hot room.
weight evenly disrupted throughout the feet.
Engage the muscles in your legs, buttocks and stomach, standing straight.
To engage your thighs and buttocks: squeeze your glutes together, then imagine you are pulling your knee caps up and squeeze your thighs together. At the same time engaging your core by concentrating on your abdomen. Feel like you are pulling your bellybutton inwards, toward your spine and drawing up from your pelvic floor (imagining you wish to stop yourself from going to the toilet!).
Palms together, interlace your fingers and thumbs, placing your knuckles under your chin. If possible arms together up to the elbows. However, if when doing this your back rounds and your shoulders come forward, open you arms enough to prevent this from happening. Shoulders drawn down and your back straight through the exercise. Keep the knuckle to chin connection throughout the entire exercise.
Keeping your mouth closed, inhale deeply through your nose using your throat to pull the air in. This will make a gentle snoring sound at the back of your throat.
Breathe in slowly and steadily for 6 counts, filling your lungs to capacity.
As you gently fill your lungs, slowly raise your elbows directly up, creating an arc in the air.
The goal is to eventually be able to raise your elbows up until your forearms touch with your ears. This can take weeks or years depending on your flexibility. Don't be disheartened if you cannot reach this final position, as long as you go to your personal edge you are getting maximum benefit from the exercise.
Your chin should stay rested into the 'u' of your hands through the 6 counts.
Immediately and in a fluid movement as soon as the 6 inhale counts have been reached, open your mouth a little and push the air out of your body slowly and steadily, making a hhhhhhhhaaaaaaaahhhhhhhh sound as the air escapes. It should resonate from your throat and make an ocean- esq. sound as it is being released.
Like you are trying to fog up a mirror.
Simultaneously, while you are exhaling for 6 counts, drop your head backwards, knuckles glued to the chin and bring your palms, wrist, forearms, and elbows together. Your elbows, wrists, arms and upturned face will form one parallel line to the ceiling.
Empty every last drop of air from your lungs and repeat roughly another 10 times, however this is really up to your teacher d'jore.
Remember keep your eyes open during the entire exercise.
Pranayama Standing Deep Breathing, will not only energise you and set you up for class, it will clear out all that old stale air and replace it with fresh oxygen, that is then zinged around your body to all the organs and muscles getting them ready and lively for class. This breathing will also help to clear and focus your mind.
If you have any questions or injuries please speak with your teacher before class.
Good for lungs and respiratory system.
Exercises nervous, respiratory and circulatory systems.


text:
Welcome to MOBicure
MOBicure is a digital health company that provides mobile technology solutions to some of the most pressing healthcare problems plaguing Nigeria and Africa.
We are a digital health company creating solutions that make use of mobile technology devices and tools such as phones, tablets, SMS, apps, Interactive Voice Recordings (IVRs) and videos to solve some of the most pressing healthcare problems facing developing countries, especially with regards to sexual & reproductive health, as well as maternal & child health. Our products are: the myPaddi & the OMOMi app.
myPaddi app provides young people with access to accurate, unbiased and youth-friendly sexual and reproductive health information, while ensuring they remain totally anonymous.
The app also connects these young people to health professionals in a real-time chat.
Omomi app provides pregnant women and mothers with access to life-saving maternal and child health information, as well as access to doctors at the touch of a button. At MOBicure, our vision is to bring healthcare to the fingertips of everyone using mobile technology.
Our core values are excellence and innovation.
Get to meet the brains behind the mypaddi and omomi projects.
Charles Akhimien
Dr Charles Immanuel Akhimien is a medical doctor and Co-founder of MOBicure. He is a UN SDG Young Leader, UN SDG Pioneer, Mandela Washington Fellow & One Young World Ambassador
Owobu Emmanuel
He is a Commonwealth Scholar, a Leap Africa Fellow and a 2017 Commonwealth Youth Awards finalist. He is currently a graduate student at the University of Edinburgh studying global e-health.
He is a Commonwealth Scholar, a Leap Africa Fellow and a 2017 Commonwealth Youth Awards finalist. He is currently a graduate student at the University of Edinburgh studying global e-health. He is particularly passionate about solving social problems using ICT. A lover of new technologies and innovative trends as well as always seeking to improve skills with every project done. He is a huge fan of The Big Bang Theory and Silicon Valley, two T.V shows that help him relax.
Fayipe Taiwo
Taiwo specializes in web development,Application Development using Java and Mobile App development (Native/Hybrid) with major focus in front-end development.
Duru Cynthia
Duru Cynthia is a Software Engineer, proficient in DevOps, Strong quality assurance professional with a degree in Computer Science
Adele Clement
Adele Clement is a Brand Developer and Strategist who likes to build future driven brands
Nwizu Onyedikachi
Nwizu Onyedikachi Doris is a graduate of Foreign Languages from the University of Benin, Nigeria. Onyedikachi works with MOBicure as the Head of Operations. She is very passionate about public health.
Eghelen Melody
Eghelen Melody is a Graphic designer for MOBicure, with some years of experience in the industry.
Ose-Iyoha Brian
Ose Iyoha Brian is a software developer. He loves all things JavaScript. But basically loves to build stuff that solves a problem irrespective of the technology involved.
Ejonavi Jerry
Jerry is a mobile developer, passionate about building scalable solutions, he develops and manage highly performant and secure web and mobile apps.....
Ojeabulu Joy
Ojeabulu Joy is the Front Desk officer for MOBicure.
© 2019 MOBicure - All Rights Reserved


text:
Home / Windows 7
Configuring User Account Control
In versions of Windows prior to Vista, many users became frustrated with the inability to perform many common tasks and therefore ran their computers with an administrative user account, often the default Administrator account created when Windows was installed. These users received total system privileges as required for installing and configuring applications, modifying system configuration, running background system tasks, installing device drivers, and performing other system configuration actions. Such a practice left the computers open to many types of attacks by malware programs such as viruses, worms, rootkits, and others.
Administrators and technical support personnel in a corporate environment were often left in a dilemma. They could grant users administrative privileges, which can result in users changing settings, either accidentally or deliberately, that disrupted computer or network performance or compromised security. Or they could limit user privileges, which often limited productivity because users were unable to perform basic tasks such as connecting to a wireless network or installing a printer driver. Beginning with Windows Vista, Microsoft addressed this problem by introducing a new feature called User Account Control (UAC). Simply put, UAC requires users performing high-level tasks to confirm that they actually initiated the task. Members of the Administrators group are logged on with only normal user privileges and must approve administrative actions before such actions will run. Nonadministrative users must provide an administrative password. Providing administrative approval to run such tasks places the computer into Admin Approval Mode.
However, the implementation of UAC in Windows Vista generated large numbers of system prompts, even for such tasks as moving, renaming, or deleting files created or modified by a different user. I frequently experienced this problem in working with my extensive collection of photographic images, many of which had been originally copied onto an older computer running Windows XP. These annoying prompts contributed to the overall low consumer satisfaction of Vista and led many people to disable UAC completely, thereby negating its advantages. Microsoft has improved UAC in Windows 7, making it more user-friendly and less annoying while still providing protection against undesirable activities (such as installing unwanted software, unwittingly installing malware, and so on). You can now configure UAC to manage the extent of prompts provided, as will be discussed in the sections to come.
For additional information on UAC improvements in Windows 7, refer to "What's New in User Account Control" at http://technet.microsoft.com/en-us/library/dd446675(WS.10).aspx.
Features of User Account Control
UAC requests approval before running administrative tasks on the computer. UAC redefines what a standard user is permitted to do. Such a user can perform many basic functions that pose no security risk; these functions previously required a user to have administrative privileges. In addition, UAC facilitates the act of providing administrative credentials when users need to perform higher-level tasks, such as installing an application or configuring system settings. Furthermore, UAC makes administrative accounts safer by limiting the types of tasks that can be performed without users providing additional consent. UAC still requests consent before allowing users to perform tasks that require higher privileges, such as system tasks.
Under UAC, all users (administrative or not) can perform the following tasks without supplying administrative credentials:
Viewing the system clock and calendar and configuring the time zone (but users cannot change the system time)
Modifying power management settings
Installing printers and hardware devices that an administrator has allowed using Group Policy
Interfacing portable devices (such as Bluetooth) with the computer
Using Wired Equivalent Privacy (WEP) to connect to approved wireless networks
Creating and configuring approved virtual private network (VPN) connections
Installing ActiveX controls from sites that an administrator has approved
Installing critical updates from Windows Update
The tasks summarized here are similar to those granted to members of the Power Users group in Windows versions prior to Vista. Windows 7 includes the Power Users group solely for backward compatibility purposes. You do not need to add users to this group to perform these functions. Add users to this group only if required for running noncertified or legacy applications. To grant this group all the privileges provided in Windows XP, you must apply a default security template that modifies default permissions on system folders and the Registry.
When authenticating a member of the Administrators group, Windows 7 issues two access tokens:
A full administrator token:
The administrator token is used only when administrative privileges are required.
A standard user token:
The standard token is used for all actions that do not require administrative privileges.
Windows 7 also marks tasks and programs as belonging to one of two integrity levels, which are implied levels of trust in these actions:
Low integrity:
A task or application (such as a web browser, email, or word processing program) that is less likely to compromise the operating system.
High integrity:
An action that performs tasks (such as installing applications) that have a higher potential for compromising the system. Applications running at low integrity levels cannot modify data in applications using a higher integrity level.
Windows 7 informs you when a task requires elevated (administrative) privileges by displaying shield icons such as those that appear in the left column of the System applet. On selecting one of these tasks, you receive a UAC prompt. Click Yes to proceed with the task or No to cancel it. When you selected one of these tasks on a Windows Vista computer, the screen dimmed and a UAC prompt (also known as an elevation prompt) displayed. When you accepted the prompt, the administrative access token granted you elevated privileges, enabling you to perform the task you have selected. In Windows 7, this behavior depends on the UAC setting you've specified (as you will learn later in this section). The default setting enables administrators to perform most of the actions marked with shield icons without receiving UAC prompts; they receive prompts for performing tasks such as installing programs or running the Registry Editor or other programs that have a high potential for producing damaging effects.
The dimmed screen indicates that the UAC prompt is running in secure desktop mode (such as when the Ctrl+Alt+Delete prompt appears when logging on to a domain-based computer). This means that you must either approve or cancel the UAC prompt before you can continue performing any other task on the computer.
A user who is not a member of the Administrators group receives only the standard user token when access is authenticated. Such a user receives the UAC prompt, which requires that a password for an Administrator user account be entered. By default in Windows 7, a nonadministrative user receives this prompt for any action marked by a shield icon.
When you receive a UAC prompt, always ensure that the action that launches the prompt is the one you want to perform. This is especially true if a UAC prompt appears unexpectedly, which could indicate a malware programattempting to run. Should this happen, click No and the program cannot run. You should then scan your computer with one or more malware detection programs.
If a background application that is minimized to the taskbar requires elevated privileges, the UAC prompt appears on the taskbar and blinks to draw attention. An example of where this would happen is in the downloading of an application from the Internet. When the download completes and approval for installation is required, the user can click the prompt to approve it. This enables the user to continue performing other tasks, such as reading email, while the download is underway; the user can continue with these tasks without being interrupted by the dimming of her screen and a UAC prompt displaying onscreen.
Application Prompts
UAC causes some third-party applications to display prompts when you attempt to run them. This helps to secure your computer because the prompt informs you of the program that is attempting to run so that you can verify that it is a program you really want to run. Click Yes to run the program or No to exit. The type of shield icon depends on the security risk involved in running the program:
High-risk blocked program:
Windows displays a message box with a red title bar and red shield stating This program has been blocked for your protection. Such a program comes from a blocked publisher and cannot be run under any circumstances.
Program signed by Windows:
The UAC prompt includes a blue title bar and blue and yellow shield for an administrative user. Click Yes to run the program. For a nonadministrative user, the prompt is shown. Provide an administrative password to run the program.
Unsigned program from a verified publisher:
When running with an administrative account, a program with a legitimate digital signature that includes its name and publisher will display a prompt. Click Yes to run the program. A nonadministrative user will receive a prompt that asks for an administrative password.
Unsigned program from a nonverified publisher:
If the third-party program does not have a digital signature that includes its name and publisher, the prompt that appears is a stronger caution. It uses a yellow title bar and yellow shield. Click Yes to run the program. Again, a nonadministrative user will receive a prompt that asks for an administrative password.
For more information on the various prompts that UAC can issue in Windows 7, refer to "UAC Processes and Interactions" at http://technet.microsoft.com/en-us/library/dd835561(WS.10).aspx.
Running Programs with Elevated Privileges
Microsoft has provided several means of configuring applications and tasks to run with elevated privileges. Use the following procedure to perform a task with elevated privileges:
Start the program or task displayed with a shield icon. The display dims and the UAC prompt appears.
Verify that the UAC prompt is requesting privileges for the task you're attempting to run (remember, some malware can deceive you here, so make certain the correct program or task is described in this prompt). If desired, click Show details for more information on the task.
If this is indeed the correct program or task, click Yes to start the task or application.
You can also mark an application to always run with elevated privileges. This situation might occur if the application developer has coded the program to access protected folders such as the %ProgramFiles% or %Systemroot% folders or requires access to the Registry. You can also configure a program to request administrative privileges from its shortcut properties. When you do this, the program always displays a UAC prompt when started from its shortcut. Use the following procedure to mark an application to always run with elevated privileges:
Ensure that you are logged on to the computer as a member of the local Administrators group.
If necessary, drag a shortcut to the desktop.
Right-click the shortcut and choose Properties.
On the Shortcut tab, click the Advanced button.
On the Advanced Properties dialog box shown, select the Run as administrator check box and then click OK.
Click OK to close the shortcut Properties dialog box.
If you are logged on using the default Administrator account created when you installed Windows 7, you do not receive any UAC prompts. Do not use this account except under emergency conditions. Best practices recommend that this account remain disabled; it is disabled by default in Windows 7.
In Windows 7, as already mentioned, you can configure several levels of UAC that determine whether prompts are displayed and how they appear on the screen. Click Start → Control Panel → System and Security and then select Change User Account Control settings under Action Center. Alternatively, you can type User Account Control into the Start menu Search field and then select this option from the search list. You receive the dialog box. Select from the following options, click OK, and then accept the UAC prompt that appears:
Always notify me when:
Windows displays a UAC prompt whenever you make changes to Windows settings or programs try to install software or make changes to your computer. This behavior is similar to that of Vista.
Default-Notify me only when programs try to make changes to my computer:
The default setting in Windows 7, this setting does not prompt you when you make changes to Windows settings. You are prompted on the secure desktop (that is, the desktop dims) when you perform higher-level actions, such as installing programs or accessing the Registry Editor.
Notify me only when programs try to make changes to my computer (do not dim my desktop):
Similar to the default setting, except that the desktop does not dim when a UAC prompt appears. With this setting, you can ignore the UAC prompt and continue performing tasks other than the task that is requesting approval.
Never notify me:
Disables UAC completely. This setting is not recommended; you should use it only when absolutely necessary to run a program that displays the red shield icon mentioned earlier in this section.
User Account Control Policies
Microsoft has provided a series of policies in Windows 7 Group Policy that govern the behavior of UAC. These policies are available from the Group Policy Management Editor snap-in or from the Local Security Policy snap-in in (available by typing Local Security from the Start menu Search field and then selecting the program from the list that appears). Use the following procedure to configure UAC policies:
Click Start, type gpedit.msc in the Search field, and then press Enter.
Navigate to the Computer Configuration\Windows Settings\ Security Settings\Local Policies\Security Options node.
Scroll to the bottom of the policy list to view and configure the available policies.
To configure a policy, right-click it and choose Properties. Choose Enabled or Disabled as required and click OK. Two of the policies offer options from a drop-down list. You can also click the Explain tab for further information on each policy.
When finished, click OK.
You can use this procedure to configure the following UAC policies:
Admin Approval Mode for the Built-in Administrator:
Governs the behavior of the built-in Administrator account. When enabled, this account displays the UAC prompt for all actions requiring elevated privileges. When disabled, this account runs all actions with full administrative privileges. This policy is disabled by default.
Allow UIAccess applications to prompt for elevation without using the secure desktop:
Determines whether User Interface Accessibility (UIAccess) programs can automatically disable the secure desktop with a standard user. When enabled, these programs (such as Remote Assistance) automatically disable the secure desktop for elevation prompts. When disabled, the application runs with UIAccess integrity regardless of its location in the file system. Note that UIAccess application programs and accessibility tools are used by developers to push input to higher desktop windows that require the uiAccess flag to be equal to true (that is, uiAccess=true). Also, the application program that wants to receive the uiAccess privilege must reside on the hard drive in a trusted location and be digitally signed. This policy is disabled by default.
Behavior of the elevation prompt for administrators in Admin Approval Mode:
Determines the behavior of the UAC prompt for administrative users. This policy has the following options:
Prompt for consent for non-Windows binaries:
Prompts a user on the secure desktop to select either Permit or Deny when a non-Microsoft program needs elevated privileges. Select Permit to run the action with the highest possible privileges. This option is the default setting.
Prompt for consent:
Prompts a user to select either Permit or Deny when an action runs that requires elevated privileges. Select Permit to run the action with the highest possible privileges.
Prompt for credentials:
Prompts for an administrative username and password when an action requires administrative privileges but does not display the secure desktop. When selected, administrative users receive the for nonadministrative users.
Prompt for consent on the secure desktop:
Prompts a user to select either Permit or Deny on the secure desktop when an action runs that requires elevated privileges. Select Permit to run the action with the highest possible privileges.
Prompt for credentials on the secure desktop:
Prompts for an administrative username and password on the secure desktop when an action requires administrative privileges. When selected, administrative users receive the prompt shown for nonadministrative users.
Elevate without prompting:
Enables the administrator to perform the action without consent or credentials. In other words, the administrator receives Admin Approval Mode automatically. This setting is not recommended for normal environments.
Behavior of the elevation prompt for standard users:
Determines the behavior of the UAC prompt for nonadministrative users. This policy has the following options:
Displays a prompt to enter an administrative username and password when a standard user attempts to run an action that requires elevated privileges. This option is the default setting.
Displays a prompt on the secure desktop to enter an administrative username and password when a standard user attempts to run an action that requires elevated privileges.
Automatically deny elevation requests:
Displays an Access is Denied message shown when a standard user attempts to run an action that requires elevated privileges.
Detect application installations and prompt for elevation:
When enabled, displays a UAC prompt when a user installs an application package that requires elevated privileges. When disabled, domain-based Group Policy or other enterprise-level technologies govern application installation behavior. This option is enabled by default in an enterprise setting and disabled by default in a home setting.
Only elevate executables that are signed and validated:
When enabled, performs public key infrastructure (PKI) signature checks on executable programs that require elevated privileges before they are permitted to run. When disabled, no PKI checks are performed. This option is disabled by default.
Only elevate UIAccess applications that are installed in secure locations:
When enabled, runs applications with UIAccess integrity only if situated in a secure location within the file system such as %ProgramFiles% or %Windir%. When disabled, the application runs with UIAccess integrity regardless of its location in the file system. This option is disabled by default.
Run all administrators in Admin Approval Mode:
When enabled, enforces Admin Approval Mode and other UAC policies. When disabled, all UAC policies are disabled and no UAC prompts are displayed. In addition, the Windows Security Center notifies the user when disabled and offers the option to enable UAC. This option is enabled by default.
Switch to the secure desktop when prompting for elevation:
When enabled, displays the secure desktop when a UAC prompt appears. When disabled, the UAC prompt remains on the interactive user's desktop. This option is enabled by default.
Virtualize file and registry write failures to per user locations:
When enabled, redirects application write failures for pre-Windows 7 applications to defined locations in the Registry and the file system, such as %ProgramFiles%, %Windir%, or %Systemroot%. When disabled, applications that write to protected locations fail, as was the case in previous Windows versions. This option is enabled by default.
Caution: If you disable the Run all administrators in Admin Approval Mode policy setting, you disable UAC completely and no prompts will appear for actions requiring elevated privileges. This leaves your computer wide open for attack by malicious software. Do not disable this setting at any time!
If you do disable this setting, note that the Windows Action Center will display a message from the notification area.
For information on problems you might encounter with UAC in Windows 7 and Windows Server 2008 R2, consult the links referenced in "User Account Control Troubleshooting Guide" at http://technet.microsoft.com/en-ca/library/ee844169(WS.10).aspx?ITPID=insider.
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Why Is the Triple Crown So Hard to Win Now? Because It Always Has Been
As American Pharoah prepares for his chance to become just the 12th horse in history, and first in 37 years, to win horse racing's Triple Crown, experts are lining up to explain why it has become so hard to win.
Most observers assert that today's thoroughbreds are bred for precocious speed, and not suited for either the 1 ½ mile distance of the Belmont Stakes or three tough races in five weeks.
We have analyzed 19 horses who won two legs of the Triple Crown since Affirmed last won it in 1978. (note: the article links to replays of, and contemporary commentary about, most of the relevant races).
We concluded that only four of these 19 horses failed to win the Triple Crown because they could not negotiate the distance of the Belmont Stakes or the grind of three tough races in five weeks. The remaining 15 horses lost their shot at immortality for the same reasons that have been preventing great horses from winning the Triple Crown for the past century.
Of these 19, 12 came to New York with a shot at the Triple Crown, after wins in the Kentucky Derby and Preakness. Another 7 horses in this span won the Belmont Stakes and another leg of the Triple Crown, but lost the Derby or the Preakness.
Of the 12 to falter in the Belmont Stakes, only four were clearly impacted by the distance of the Belmont and/or the Triple Crown grind: War Emblem, Funny Cide, Big Brown and California Chrome. Northern Dancer met a similar fate in 1964, as did Majestic Prince in 1969.
Seven of the remaining 8 to lose at Belmont ran very well there and suffered fairly narrow losses, arguably as a result of questionable race riding tactics. When a Triple Crown is on the line, the pressure forces a jockey to pursue pacesetters earlier than he or she might in an ordinary race. These cases show that the biggest strain of a Triple Crown chase may be the one placed on the humans associated with the horse.
In hindsight, the masterful ride of young Steve Cauthen aboard Affirmed becomes all the more impressive, and the pressure on American Pharoah's jockey Victor Espinoza, increases (although this is his third try at a Triple Crown).
A look at the 7 horses who endured questionable riding tactics:
In 1978, the trainer of Spectacular Bid, Buddy Delp told people it would take an "Act of God" for the horse to lose, and flippantly predicted that he would win by 32 lengths. He was chasing the legend of Secretariat. Spectacular Bid entered the backstretch second behind 80-1 shot Gallant Best, and jockey Ronnie Franklin sent Spectacular Bid surging to the lead. Coastal engaged him in the stretch, and Bid had nothing left.
In 1981, Pleasant Colony's jockey failed to press the slow pace of longshot Summing, reserving his energy for the seemingly more talented closers behind him. Pleasant Colony could not run down the pacesetters, and finished third.
In 1989, Sunday Silence won the Derby and Preakness over Easy Goer. In the Belmont, Le Voyageur, a regally bred foreign horse (by Seattle Slew out of Davona Dale, herself a Filly Triple Crown winner), set a strong pace. With a Triple Crown on the line, Sunday Silence's jockey Pat Valenzuela was forced to attack this unknown front runner entering the far turn; Easy Goer's Pat Day was able to lay in wait and made a single explosive run entering the stretch and pulled away to an easy victory.
In 1997, Silver Charm won the Derby in a thrilling three-way duel with Free House and Captain Bodgit, and the Preakness by a nose over Captain Bodgit. With Captain Bodgit sidelined, Silver Charm focused on outdueling Free House into the stretch of the Belmont, only to yield the lead in the final fifty yards to Touch Gold, who had the luxury of waiting till the final yards to strike.
In 1998, Kent Desormeaux prematurely sent Real Quiet to a commanding lead into the upper stretch, but was nipped at the wire by a fast-closing Victory Gallop.
In 1999, Charismatic was forced to confront the talented filly, Silver Bullet Day, through challenging fractions. He led with a furlong to go but was caught by longshot Lemon Drop Kid. An alternative view is that Charismatic's loss is more fairly attributable to an injury suffered in the race - a fate shared by Tim Tam in 1958, and Carry Back in 1961.
In 2004, undefeated Smarty Jones faced a phalanx of talented challengers. After a sensible first half mile, Smarty Jones took on Purge, the frontrunner who had a legitimate shot. Right as Smarty Jones passed Purge, he was engaged by Eddington, and then Rock Hard Ten. He ran a grueling three-quarter mile segment of the race more than a second faster than Secretariat had. Unlike Secretariat, Smarty Jones staggered in the final 70 yards and was collared by Birdstone, the only horse in the race who sat out the furious fray on the backstretch.
In 1987, Alysheba lost not to the 12 furlongs, but to New York's then-strict anti-drug rules for horses. Alysheba required the diuretic drug Lasix to perform at his best, and it was banned in New York at the time. The hot, muggy Belmont day ensured that Alysheba would not run his best, and Bet Twice won the race by nearly 15 lengths. This was the first of three losses in a row for Alysheba - all without Lasix.
Of the seven horses who won two legs of the Triple Crown, including the Belmont, since 1978:
Risen Star, Point Given and Afleet Alex were by far the best of their crop, but suffered tough trips in the Kentucky Derby, then went on to resounding victories in the Preakness and Belmont. Empire Maker and Lookin at Lucky are two others who may have won two legs after tough trips in the Derby, but elected to skip one of the legs with no Triple Crown possibility (something that Gallant Man also did in 1957). While surely tough trips in the Derby as more commonplace now that the race has a full field of 20 nearly every year, a rough Derby journey also cost the great Native Dancer a shot a Triple Crown glory in 1953, as well as Little Current in 1974. Each went on to resounding wins in the Preakness and Belmont Stakes.
Thunder Gulch, Hansel and Swale simply came up empty in the Derby or the Preakness, a fate that felled Damascus in the 1967 Kentucky Derby.
Tabasco Cat arguably lost the 1994 Kentucky Derby due to track conditions - the same fate that cost Riva Ridge the Preakness in 1972.
In 2001, Laura Hillenbrand, author of Seabiscuit, wrote in an op-ed for The New York Times,
"In 1973, the racing world was haunted by a mood similar to that felt today. A quarter century had passed since the coming of the last Triple Crown winner, Citation. Fans wondered if they would ever see another horse sweep the series. Then, on June 9, a red colossus named Secretariat roared down the Belmont homestretch on an awesome 31-length lead, winning the Triple Crown in world-record time…. Those who worry that we may never see this again should be patient. True greatness is extremely rare; the next Triple Crown winner will be worth the wait."
Will we see true greatness on June 6? I think we will.
Are you attending this year's Belmont Stakes? Read our Insider's Guide to Attending the Belmont Stakes.
Want to relive Secretariat's 1973 triumph? Read our "On This Day" article about it.
Tags: triple crown hard to win
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Opera North seeks to encourage people, of all ages and from all walks of life, to explore opera and the work of the Company and to break down perceived barriers to engagement with the artform. It seeks to champion excellence in opera and related fields by, with and for young people and to use arts participation and performance to enhance the health and wellbeing of people in the communities where we work.
Our Early Years and Family work creates shared experiences for families to enjoy together, whether attending a relaxed family-friendly performance by members of our award-winning professional orchestra and chorus, or singing together in our Little Singers programme.
Our Family initiatives provide opportunities for families of all kinds to get involved, offering one-off workshops, small-scale family shows and weekly 'early years' activities.
We develop programmes to help families play, learn, understand and express themselves together.
Find upcoming events, workshops and performances for children and families.
Our Youth Ensembles aim to develop musical excellence and inspire the next generation of singers and musicians from the North of England.
Young people are encouraged to explore their own creativity by gaining practical experience across a broad range of artforms – music, singing, instrumental, drama, direction, design and technical skills. Professional artists lead our workshops, providing stimulating leadership and guidance which can result in young people producing and performing their own work.
Youth Ensembles include Opera North Children's Chorus (ages 8-12), Opera North Young Voices (ages 12-15), Opera North Youth Chorus (ages 14-19), and Opera North Youth Company. Instrumental Ensemble opportunities include Orchestra Camp, Orchestra Academy and Junior Strings.
Explore Youth Ensembles for young people aged 8 – 19.
Opera North's Lifelong Learning strand provides inspirational arts activities for adults of all ages, led by professional artists, in a friendly and relaxed environment.
Our Sing ON programme of weekly singing groups for Over 55s encourages participants to grow their social networks, engage in regular arts activity, and be inspired by opera.
We also offer regular After Hours evening courses for adults of all ages to explore singing and the work of Opera North.
Find out more about current activities for adults.
We support teachers and schools to deliver the curriculum in music and drama, creating extraordinary experiences in schools across the North of England.
Our Schools work reaches large numbers of children and young people, delivering a high quality first experience of opera through Schools' Matinee performances, the creation of work especially for a young audience, and connecting with our award-winning professional Chorus and Orchestra.
Visit our dedicated Schools page for more information, online resource packs, and details of upcoming activities we can offer to schools.
In Harmony Opera North is an award-winning Community Residency programme involving four primary schools in south Leeds and working with over 1700 children and young people on a weekly basis.


Kim Vo blogs about the second episode and explains the best way for a client to approach a stylist.
They are the toughest clients to work for. They had their signature looks and they were going to be tough to change. Sometimes the littlest choice will make the biggest statements.
Yes -- we were debating who took a right or a wrong risk. Parker did take a risk that I was happy about, but it was the wrong choice. It was not age-appropriate. He dyed it jet black and gave her bangs. He should have softened it with a chestnut color with some highlights and kept its deep rich color. No one should be that dark after their teenage years.
Do you have any parting words for Parker?
Parker, many times in my life, I "faked it till I made it." Confidence is key, but also the proof is in the pudding. I like your confidence but I want to see the work that backs it up.
Any thoughts about how you should approach a stylist with what you want, opposed to what looks good?
A trick that I have is that I play devil's advocate by telling them that what they need is textbook and then I tell them what I think they need. As a client you need to connect with your stylist.
I think Charlie has put himself on the map.


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CID probe into Sri Lankan women sold for sex in Oman
COLOMBO – Sri Lanka's Criminal Investigations Department is investigating a human trafficking ring where Sri Lankan women who travel to Oman to work as domestic aides are auctioned and sold to work as escorts.
Police Spokesperson & Attorney SSP Nihal Thalduwa told News 1said that a team of detectives led by two ASPs visited Oman to conduct the investigation.
Multiple foreign employment agencies are sending young girls and women to Oman promising them employment as domestic aides, and most of them had used tourist visas to enter the middle-eastern country.
Detectives uncovered that when the group reaches Oman, they are lined up according to their age and appearance and auctions and sold to engage in sexual activity.
"I was sexually abused inside a room. I am not afraid to reveal this to the CID and the Foreign Employment Bureau. I went to every single place possible in search of justice. I told the person that I was groped while cleaning, and she told me that it's normal, and that I need to be prepared for all these things. She also offered me money to be with her. When I told the embassy about it and requested to return to the country, they refused to accept me because I was on a visit visa. I am only 21 years old, and this is my first time here. I came here because I am helpless," said one Sri Lankan woman who was sexually abused in oman.
Nissanka Priyadarshana is the man who exposed the trafficking ring and he told News 1st that some agencies in Sri Lanka send many young girls to Oman, and in Oman, the agencies line up the girls and sell them for money.
He urged that people not go to fake agencies and fall into trouble.
SSP Nihal Thalduwa, the country's Police Spokesperson said that agencies are preparing the passports and tourist visas to send the women abroad and the investigations revealed that there are brokers in Sri Lanka and Oman.
The Sri Lanka Bureau of Foreign Employment said that necessary action will be taken after the CID investigation report is made available.
The Sri Lankan Embassy in Oman told News 1st that among the 90 plus women houses at a safe-house operated by the Embassy in Oman, some were victims of this smuggling operation.
The Embassy said that it completely supported the investigations carried out by the CID. (News 1st)
UK Trade Envoy for Sri Lanka is in Colombo
WEATHER FORECAST: Issued at 05.30 a.m. on November 10


Microelectronics - Instrumentation - Flow Controller - Semitorr Group Inc.
SemiTorr Group, Inc. is your stocking distribution partner supporting Dwyer in many states across the US. Dwyer is an industry leader in instrumentation including pressure, temperature, flow control and level control. For more information on Dwyer, please visit https://www.dwyer-inst.com/ or contact your local SemiTorr representative.


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Main Stage Saturday 1 July
Oscar and the Wolf
Straight from hell into heaven. That's what happened to Oscar and the Wolf last summer. In June, Max Colombie had to postpone part of his European tour because of mental health issues, but a few weeks later, he gave majestic performances at Lowlands and Pukkelpop. The arrangements. The comprehensive accompaniment. The integration of songs from the new 'The shimmer' (2022), which Max himself called a "stroll through four seasons". All of it was of the highest quality; an Olympic-level achievement. The concerts were brought to a close with a new song: 'Warrior' can be heard as far as the offices of the Royal Belgian Football Association in Brussels. An overflowing Sportpaleis in November is proof that Oscar and the Wolf has found the perfect balance between electronica and emotion. At least, insofar as it didn't exist yet…
Thu 29 June Main Stage
Thu 29 June KluB C