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Lukasiewicz, Peter D. and Colleen R. Shields. Different combinations of GABAA and GABAC receptors confer distinct temporal properties to retinal synaptic responses. J. Neurophysiol. 79:3157–3167, 1998. This study addresses how γ-aminobutyric acid-A(GABAA) and GABAC receptors confer distinct temporal properties to neuronal synaptic responses. The retina is a model system for the study of postsynaptic contributions to synaptic responses because GABAergic amacrine cells synapse onto neurons, which have different combinations of GABAA and GABAC receptors. It is not known, however, how GABAA versus GABAC receptors influence the time course of retinal synaptic responses or what proportion of inhibitory input is mediated by each receptor type. We examined the time courses of synaptic responses mediated by GABA receptors in ganglion and bipolar cells by recording currents evoked by activating amacrine cells with a stimulating electrode in the salamander retinal slice. The pharmacologically isolated, GABAergic synaptic currents were long-lasting in bipolar cells and relatively brief in ganglion cells. The receptors that mediated these temporally distinct synaptic responses exhibited different pharmacological properties. In ganglion cells, GABAergic synaptic currents were abolished by the GABAA receptor antagonists bicuculline or SR95531. In bipolar cells, the GABAC receptor antagonist 3-aminopropyl[methyl]phosphonic acid (3-APMPA) largely blocked GABAergic synaptic responses; the remaining response was blocked by bicuculline or SR95531. The GABAA receptor component of the bipolar cell response was relatively brief compared with the GABAC receptor component. Puffing GABA onto ganglion cell dendrites or bipolar cell terminals yielded similar pharmacological and kinetic results, indicating that transmitter release differences did not determine the response time courses. Moreover, the GABAC receptors on bipolar cells may be different from those reported in rat or fish retina because imidazole-4-acetic acid (I4AA), which acts as an antagonist in these preparations, acts as an agonist in salamander. Our data show that the prolonged synaptic responses in bipolar cells were mediated predominantly by GABAC receptors, whereas transient synaptic responses in ganglion cells were mediated by GABAA receptors.
In the CNS the inhibitory transmitter γ-aminobutyric acid (GABA) interacts with two types of ionotropic receptors, GABAA and GABAC. The GABA receptor complement of specific neurons is important because there are differences in the properties of GABAA and GABAC receptors that may have significant functional consequences. These receptor types may determine the time course of synaptic responses. Native GABAA and GABAC receptors (Qian and Dowling 1993, 1995) as well as GABAA and ρ receptor subunits expressed in Xenopus oocytes (Amin and Weiss 1994) show different time courses in response to GABA application. GABAC receptor–mediated responses show minimal desensitization and slow deactivation, whereas GABAA receptor–mediated responses show marked desensitization and rapid deactivation. However, it is not known whether these differences in receptor properties give rise to differences in time courses of synaptic responses.
GABA is an important inhibitory neurotransmitter in the vertebrate retina that modulates synaptic transmission at both the outer and inner plexiform layers (Chun and Wässle 1989; Lukasiewicz and Werblin 1994; Marc et al. 1978; Murakami et al. 1982; Pourcho and Owczarzak 1989; Wu 1991). There is an abundance of GABAC receptors in the retina, making it a good system for studying their synaptic function. In other parts of the CNS, the roles of GABAC receptors are poorly understood.
Because GABAC and GABAA receptors are differentially expressed on distinct classes of retinal neurons, we can determine how they might shape different types of synaptic responses. Electrophysiological studies indicate that GABAC receptors are found on horizontal cells of teleost fish (Dong et al. 1994; Qian and Dowling 1993) and on bipolar cells in both mammals and cold-blooded vertebrates (Feigenspan et al. 1993; Lukasiewicz et al. 1994; Lukasiewicz and Wong 1997; Matthews et al. 1994; Qian and Dowling 1995). In ganglion and amacrine cells, GABA-evoked responses appear to be mediated either predominately or exclusively by GABAA receptors (Dong et al. 1994; Feigenspan et al. 1993; Lukasiewicz et al. 1994; Lukasiewicz and Wong 1997). However, other studies report that ganglion cells in tiger salamander and turtle possess a minor component of GABAC receptors and a major component of GABAA receptors (Liu and Lasater 1994; Zhang and Slaughter 1995). In agreement with most of the physiological studies, immunocytochemical (Enz et al. 1996; Koulen et al. 1997) and in situ hybridization (Enz et al. 1995; Qian et al. 1997; but see Albrecht and Darlison 1995) studies show that ρ-subunits are found mainly on bipolar cells in mammals, fish, and birds.
In this study we have characterized the time courses of synaptic inhibition mediated primarily by GABAC receptors on bipolar cells and by GABAA receptors on ganglion cells in the tiger salamander retinal slice. We evoked synaptic responses that were mediated by GABAA and GABAC receptors by eliciting GABA release from amacrine cells with electrical stimulation in the inner plexiform layer (IPL). The GABA receptor–mediated components of the synaptic response were measured after blocking glutamate and glycine receptor function. Inhibitory synaptic responses in ganglion cells were relatively transient compared with those in bipolar cells. The results from our pharmacological studies indicated that the relatively prolonged synaptic responses in bipolar cells were mediated mainly by GABAC receptors. Small transient GABAA receptor–mediated responses were often seen in bipolar cells after GABAC receptor blockade. In addition, the salamander bipolar cell GABAC receptors were atypical in that they were activated by the GABAC receptor antagonist imidazole-4-acetic acid (I4AA) (Qian and Dowling 1994, 1995; Pan and Lipton 1995). Pharmacological results indicated that the transient GABAergic synaptic responses in ganglion cells, by contrast, were mediated exclusively by GABAA receptors. These results suggest that different GABA receptors contribute to the distinct time courses of inhibitory synaptic responses in bipolar and ganglion cells.
Whole cell patch recording in tiger salamander retinal slices
Whole cell patch recordings (Hammill et al. 1981) were made from bipolar cells and ganglion cells in retinal slice preparations (Werblin 1978). The preparation of the retinal slices and the recording procedures have been described in detail previously (Lukasiewicz and Roeder 1995; Lukasiewicz and Werblin 1994). Tiger salamanders were obtained from C. D. Sullivan (Nashville, TN) and kept at 5°C on a 12 h light:12 h dark cycle. Experiments were performed at room temperature (19–22°C).
Electrode and bath solutions
The intracellular electrode solution consisted of (in mM) 95.25 cesium gluconate, 8 tetraethylammonium chloride (TEA), 0.4 magnesium chloride, 11 ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA), and 10 sodium N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES), adjusted to pH 7.7 with cesium hydroxide. The bathing medium (salamander Ringer) contained (in mM) 112 sodium chloride, 2 potassium chloride, 2 calcium chloride, 1 magnesium chloride, 5 glucose, and 5 HEPES, adjusted to pH 7.8 with NaOH. The calculated E Cl for these intra- and extracellular solutions was −65 mV. Unless otherwise indicated, all chemicals were obtained from Sigma Chemical (St. Louis, MO). 6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX), 2-(3-carboxypropyl)-3-amino-6-(4-methoxyphenyl)pyridazinium bromide (SR-95531), and 3-aminopropyl[methyl]phosphonic acid (3-APMPA) were obtained from Research Biochemicals (Natick, MA), and d-2-amino-5-phosphonopentanoic acid (d-AP5) was obtained from Precision Biochemicals (Vancouver, BC). In some cells, responses were recorded using the gramicidin, perforated-patch technique (Ebihara et al. 1995). The perforated-patch electrode solution consisted of the following (in mM): 107.5 cesium chloride and 10 HEPES, adjusted to pH 7.5 with Tris base. Gramicidin stock solution was prepared by dissolving 10 mg/ml in methanol and was diluted to its final concentration of 50 or 100 μg/ml with the electrode solution. The electrode tip was filled with gramicidin-free electrode solution to enhance seal formation. No differences in response were observed when whole cell versus perforated-patch techniques was used.
Membrane potential values given in this paper were corrected for junction potentials. Liquid junction potentials were determined as described by Fenwick et al. (1982). They were typically −11 and −2 mV for the cesium gluconate and perforated-patch electrode solutions, respectively.
The control bathing solution used in our slice experiments was formulated to pharmacologically isolate the GABA receptor–mediated component of inhibitory postsynaptic currents (IPSCs). Glycinergic inhibitory synaptic responses were blocked with strychnine (2 μM) and N-methyl-d-aspartate (NMDA), and non-NMDA receptor–mediated responses were blocked with d-AP5 (50 μM) and CNQX (10 μM), respectively (Belgum et al. 1984; Diamond and Copenhagen 1993; Mittman et al. 1990). Voltage-gated potassium currents were blocked by including cesium and TEA in the recording electrodes (Lukasiewicz and Werblin 1988). Antagonists were applied over a relatively large area of the slice under study (several mm in width) by a gravity-driven superfusion system as described previously (Lukasiewicz and Roeder 1995).
Puffing agonists onto ganglion cell dendrites and bipolar cell terminals
GABA agonists were puffed onto the dendrites of ganglion cells or the terminals of bipolar cells in the slice preparation with a Picospritzer (General Valve, Fairfield, NJ) at 45- or 60-s intervals. The puff pressure and/or duration were adjusted to give a half-maximal or smaller response to the GABA receptor agonists. Also, because the puff electrode could not be placed directly onto the cells' processes and the slice was continuously superfused (diluting the puff), the GABA concentration at the receptors was most likely significantly less than the pipette concentration.
Bipolar electrode stimulation of amacrine cell inputs
Amacrine cell inputs to bipolar and ganglion cells were elicited by electrical stimulation in one of two ways. Synaptic inputs were elicited with an extracellular patch electrode filled with salamander Ringer (Zhang and Trussell 1994) that was placed in the IPL. A silver/silver chloride electrode (separate from the bath ground) placed near the slice served as return path for the stimulating current. In some cases, a custom concentric bipolar electrode (MCE-100, Rhodes Medical Instruments, Woodland Hills, CA), placed in the IPL, was used to elicit synaptic inputs. The distal part of the electrode shaft (70 mm) was 150-μm diam, and it tapered to a center contact of 25 μm with an outer contact of 100 μm. The stimuli were generated by either a Grass S48 or a Grass SD9 stimulator (West Warwick, RI) that was triggered by the data acquisition program. The duration and magnitude of the stimuli were adjusted to minimize synaptic fatigue and to elicit reproducible responses (typically 0.1–3 ms duration, 1–7 V). With the extracellular patch electrode, stimuli were typically shorter in duration (0.05–1 ms). Similar results were obtained with both methods. We could not test for temporal summation of synaptic GABA responses because interstimulus intervals of <90 s resulted in depression of the subsequent response, possibly due to presynaptic fatigue. Two observations indicated that the stimulation was confined to the IPL and was focal. First, we were never able to elicit responses from bipolar cells without axon terminals. (Some cells were filled with Lucifer yellow to image their terminals.) Second, the stimulating electrode had to be placed in the same IPL sublamina as the bipolar terminals to evoke a response. Responses were never elicited from presumed on or off bipolar cells (as determined from the stratification of their terminal processes) when the stimulating electrode was placed in the off or on sublamina, respectively (i.e., opposite IPL sublamina).
Recording system
The microscope system and patch-clamp apparatus used for this study were described in Lukasiewicz and Roeder (1995). Electrodes were pulled from borosilicate glass (TW150F-4, W.P.I., Sarasota, FL) with a Sachs-Flaming puller (Sutter Instruments, Novato, CA) and had measured resistances of <5 MΩ. The measured series resistances were typically 15 MΩ (read from Axopatch 200B series resistance dial) and were usually compensated by 90%. Patchit software (Geo. Grant, Somerville, MA) was used to generate voltage command outputs, acquire data, and trigger the Picospritzer and the stimulator. The data were digitized and stored with a 486-PC using a Labmaster DMA data acquisition board (Scientific Solutions, Solon, OH). Responses were filtered at 1 kHz with the four-pole Bessel low-pass filter on the Axopatch 200B and sampled at 0.5–2 kHz. Data were analyzed using Tack software (Geo. Grant, Somerville, MA). Time constants for current decays (from 90% of peak current amplitude) were determined by fitting a three parameter single exponential equation (Y = A + Be −CX) to the response using the Marquardt-Levenberg algorithm. Results are expressed as means ± SE.
GABAergic IPSCs are brief in ganglion cells and long-lasting in bipolar cells
Here we examine the time courses of synaptic responses thought to be mediated predominantly by either GABAA or GABAC receptors. Figure 1 A shows a GABA receptor–mediated IPSC that was recorded from a ganglion cell in response to an electrical shock in the IPL. The GABA receptor–mediated component of the synaptic responses was isolated by blocking glycine and glutamate receptors as described in methods. Cells were voltage clamped to 0 mV to minimize the driving force for nonspecific cation responses and maximize the driving force for chloride-mediated, GABA responses (calculated E Cl = −65 mV). In Fig. 1 B the response is replotted on a shorter time scale. The ganglion cell IPSC reached its peak amplitude in 6 ms and then decayed rapidly. The falling phase of the response could be fit with a single exponential with a time constant of 37 ms. On average, the ganglion cell response time-to-peak amplitude was 5.0 ± 0.3 (SE) ms and decayed with a time constant of 41 ± 4 ms (n = 13). The same electrical shocks evoked IPSCs in bipolar cells that were much more prolonged than those measured in ganglion cells. Figure 1 A also shows an example of a GABA receptor–mediated IPSC recorded in a bipolar cell. For comparison, the bipolar cell IPSC is scaled to the same amplitude as the ganglion cell IPSC. The bipolar cell synaptic response reached its peak amplitude in 245 ms, much more slowly than the ganglion cell response. In comparison to the ganglion cells, the bipolar cell responses decayed minimally over the time course of the recording. The decay of the bipolar response was fit with a single exponential with a time constant of 1,351 ms (Fig. 1 A). On average, the bipolar cell response decayed with a time constant of 1,918 ± 163 ms, and its time-to-peak amplitude was 288 ± 44 ms (n = 15). The differences in response time courses could be due to either a difference in GABA receptor complements or to different time courses of GABA release onto ganglion and bipolar cells.
Responses evoked by puffs of GABA onto bipolar cells or ganglion cells show similar differences in time course (Fig. 1 C). The ganglion cell puff-evoked responses were relatively transient, whereas the bipolar cell puff-evoked responses were more prolonged. The falling phase of the bipolar response to a GABA puff could be fit with a single exponential with a time constant of 872 ms (Fig. 1 C). The average time constant for the bipolar cell response decay was 1,038 ± 157 ms (n = 15). The ganglion cell response to an identical GABA puff decayed more rapidly; its falling phase could be fit with a single exponential fit with a time constant of 77 ms (Fig. 1 D). The average time constant for the ganglion cell response decay was 133 ± 24 (n = 9). The GABA puff results indicate that receptor complement contributes to differences in IPSC time course independent of presynaptic GABA release mechanisms.
Only GABAA receptors mediated GABAergic IPSCs in ganglion cells
To determine the complement of GABA receptors that mediated IPSCs on ganglion cells, we evoked IPSCs in the presence or absence of the GABAA receptor antagonist bicuculline. The GABA receptor–mediated IPSC in the ganglion cell illustrated in Fig. 2 A was completely blocked by bicuculline (100 μM). The responses in Fig. 2 B, expressed as charge-transfer, were measured by integrating the currents. In all ganglion cells tested, the GABAergic IPSC was always completely (1.6 ± 0.9% of control, n = 14) and reversibly blocked by bicuculline. These data indicate that GABAergic synaptic currents are mediated exclusively by GABAA receptors in ganglion cells. These results are in agreement with other studies showing that the GABA-evoked currents in third-order retinal neurons are completely blocked by bicuculline (Dong et al. 1994; Feigenspan et al. 1993; Lukasiewicz and Wong 1997; Yeh et al. 1996).
Pharmacological properties of GABA-evoked currents in ganglion cells
It was recently reported that ∼20% of the GABA-elicited current recorded in tiger salamander ganglion cells (the SR-95531–resistant component) was mediated by GABAC receptors (Zhang and Slaughter 1995). We puffed GABA onto the dendrites of ganglion cells (100–250 μM pipette concentration) after blocking glycine and glutamate receptors. Figure 3 A shows that the GABA-elicited current in the ganglion cells was completely blocked by bicuculline, indicating that this response was mediated exclusively by GABAA receptors. We found that, at best, GABAC receptors underlie only a very minor part of the GABA-elicited response in tiger salamander ganglion cells. On average, the GABA-elicited charge transfer was reversibly reduced to2.6 ± 0.6% (n = 17) of control by bicuculline (100–200 μM; Fig. 3 B).
The lack of GABAC receptor–mediated responses in our ganglion cells may have been due to the wash out of an intracellular component essential for GABAC responses. Therefore we performed recordings using gramicidin-perforated patches. Results similar to our whole cell recordings were obtained using gramicidin-filled pipettes and SR-95531 (7 or 20 μM) to block GABAA receptors. The SR-95531–resistant component of the GABA-elicited currents comprised 3 ± 0.9% (n = 6) of the control charge transfer. These data confirm that the currents evoked by GABA puffs were mediated almost entirely by GABAA receptors.
Both GABAC and GABAA receptors mediated GABAergic IPSCs in bipolar cells
GABA-gated chloride currents in tiger salamander bipolar cells are mediated by both GABAC and GABAA receptors (Lukasiewicz et al. 1994; Lukasiewicz and Werblin 1994). Here we characterize the actions of the GABAC receptor antagonist 3-APMPA (Pan and Lipton 1995; Woodward et al. 1993) on GABAergic IPSCs and GABA-evoked currents.
To determine the contribution of GABAC receptors to IPSCs in bipolar cells, we evoked IPSCs in the presence or absence of GABAA receptor antagonists. Figure 4 A shows a typical GABA receptor–mediated bipolar cell response evoked by synaptic activation of amacrine cells. Bath application of bicuculline or SR-95531 caused a small reduction of the synaptic response. On average, bicuculline (150 or 200 μM) or SR-95531 (15 μM) reduced the GABAA receptor–mediated synaptic charge transfer in bipolar cells to79 ± 6% (n = 14) or 75 ± 11% (n = 4) of the control responses, respectively (Fig. 4 C). These data are consistent with GABAC receptors mediating a major fraction of the bipolar cell IPSC.
In three bipolar cells, bicuculline increased the GABAergic IPSC charge transfer 1.40 ± 0.08-fold. The enhancement of the bipolar IPSC by bicuculline may have been due to disinhibition of GABAergic amacrine cells, which possess primarily GABAA receptors (Feigenspan et al. 1993; unpublished observations).
To confirm that the bicuculline-resistant component of the bipolar IPSC was indeed mediated by GABAC receptors, we determined whether or not this component was sensitive to the GABAC antagonist, 3-APMPA. Figure 4 A shows that the SR-95531–resistant component of the IPSC was completely blocked by 3-APMPA (500 μM), confirming that this component of the response was mediated by GABAC receptors. In all bipolar cells tested, the GABA receptor–mediated IPSCs were completely blocked by the combination of 3-APMPA and bicuculline (3 ± 1% of control, n = 8) or SR-95531 (2.4 ± 1.4% of control, n = 4; Fig. 4 C).
Time course of the GABAA receptor component of bipolar cell IPSC was brief
The majority of the GABAergic IPSC in bipolar cells was mediated by GABAC receptors, whereas the remaining minority of the response was mediated by GABAA receptors. Subtracting the bicuculline- or SR-95531–resistant component from the control response derived the GABAA receptor component of the bipolar cell IPSC. Figure 4 A shows an example of a GABAA receptor–mediated IPSC determined by subtraction, which is compared with the GABAC (or SR-95531–resistant) component of the response. The GABAA component of the IPSC decayed more rapidly than theGABAC component and was fit by a single exponential decay with a time constant of 125 ms (169 ± 20 ms; n = 8). The decay time course of GABAC component (τ = 1,320 ms), measured in the presence of SR-95531, was similar to the time course of control bipolar cell IPSC noted above inFig. 1.
Similar time course results were obtained when theGABAA receptor–mediated component of the bipolar cell IPSC was isolated by blocking GABAC receptors with 3-APMPA. Figure 4 B shows a bipolar cell IPSC recorded in the absence or presence of 3-APMPA (500 μM). The response in this cell was almost completely blocked by 3-APMPA, indicating that the IPSC was mediated primarily by GABAC receptors. In all bipolar cells tested, 3-APMPA reduced the synaptic charge transfer to 7.1 ± 1.5% of control (n = 12). Bicuculline blocked the 3-APMPA–resistant component (1.3 ± 0.9% of control, n = 8), confirming that it was mediated by GABAA receptors (Fig. 4 D). The 3-APMPA–resistant component of the response exhibited a relatively brief time course with a rapid time-to-peak of 12 ms (9 ± 2 ms, n = 5) and a relatively fast decay, which could be fit by a single exponential with a time constant of 146 ms (171 ± 42 ms, n = 5; Fig. 4 B). The time courses of these responses were similar to those derived above by subtraction.
Pharmacological properties of GABA-evoked currents in bipolar cells
To determine the types of GABA receptors that mediated the bipolar cell response illustrated in Fig. 1 C, GABA was puffed at the terminals of bipolar cells (200 μM pipette concentration) after glycine and glutamate receptors were blocked. Figure 5 A shows that GABA-elicited currents had two components, a small bicuculline-sensitive current and a large bicuculline-insensitive current that are thought to be mediated by GABAA and GABAC receptors, respectively. The bicuculline-insensitive component of the response was on average 82 ± 4.5% (n = 7) of the control response (Fig. 5 C), indicating that GABAA receptors mediated ∼20% of the total response, confirming similar findings by others(Lukasiewicz et al. 1994; Wellis and Werblin 1995). Inthe presence of bicuculline (100 or 150 μM), 3-APMPA (500 μM) completely blocked the GABA-elicited current, confirming that the bicuculline-insensitive component of the response was mediated by GABAC receptors. In seven of seven bipolar cells, the coapplication of bicuculline and3-APMPA completely and reversibly blocked (2.1 ± 0.7% of control charge transfer, n = 7) the GABA-elicited responses (Fig. 5 C).
To examine the time course of the GABAA receptor–mediated component of the GABA-evoked response, we blocked the GABAC receptors with 3-APMPA (500 μM). An example of a 3-APMPA–resistant, GABAA receptor–mediated response is shown in Fig. 5 B. GABAC receptors mediated most of the response in this cell, as evidenced by the large reduction by 3-APMPA. On average, 3-APMPA reduced the charge transfer to 22 ± 4% of control (n = 16; Fig. 5 D). The 3-APMPA–resistant component of the response was blocked by bicuculline (to 2.9 ± 0.9% of control, n = 16), indicating that it was mediated byGABAA receptors (Fig. 5 D). The time course of the3-APMPA–resistant, GABAA receptor component decayed more rapidly than the control time course and was fit by a single exponential with a time constant of 176 ms(248 ± 22 ms, n = 16). The control time course, in contrast, was fit by a single exponential with a time constant of 709 ms, which was similar to the average control time constant noted above (1,038 ms). The GABAA component of the puff-evoked response decayed somewhat more slowly than the puff-evoked response in ganglion cells (see Fig. 1 D).
In teleost horizontal cells (Qian and Dowling 1994) and in both rat and teleost bipolar cells (Pan and Lipton 1995; Qian and Dowling 1995), I4AA has been reported to reduce or block responses mediated by GABAC receptors. Because I4AA is also an agonist at GABAA receptors, its effects at these receptors must be blocked with bicuculline before determining its actions at GABAC receptors. We found that in the presence of bicuculline, bath-applied I4AA elicited a relatively prolonged outward current when cells were held at 0 mV. The pharmacology of the receptors mediating the I4AA current was determined by puffing I4AA onto bipolar cell terminals in the presence or absence of GABA receptor antagonists. Figure 6, A and B, shows that the I4AA-elicited current was relatively insensitive to bicuculline antagonism. In the presence of bicuculline (150 or 250 μM), the I4AA response was94 ± 4% (n = 6) of the control response charge transfer, indicating that the response was not mediated significantly by GABAA receptors. Picrotoxin (150 μM), which acts at both GABAA and GABAC receptors, by contrast, completely and reversibly blocked (3 ± 0.9% of control, n = 7) the I4AA-elicited current (Fig. 6, A and B). These data indicate that I4AA acts as an agonist, and not as an antagonist, at GABAC receptors on the terminals of tiger salamander bipolar cells.
Puffs of I4AA (500 μM pipette concentration) also elicited responses in ganglion cells. In contrast to its effects at bipolar cells, I4AA-evoked currents were completely and reversibly blocked by bicuculline (100 μM; 3.0 ± 0.6% of control, n = 8) as shown in Fig. 6, C and D. This indicates that I4AA acts as an agonist at GABAA receptors on ganglion cells and that the I4AA-sensitive, GABAC receptors found on bipolar cells are not present on ganglion cells.
Do the properties of postsynaptic GABA receptors contribute to IPSC time courses?
Studies with both isolated retinal neurons and withρ-subunit receptor expression systems have shown that responses mediated by GABAA and GABAC receptors have different time courses (Amin and Weiss 1994; Qian and Dowling 1993, 1995). Here, we show the first characterization of pharmacologically isolated, GABAergic synaptic responses in the vertebrate retina. Our results suggest that the time courses of ganglion cell and bipolar cell IPSCs are determined, in part, by different complements of GABA receptors. GABAergic synaptic responses in bipolar cells and ganglion cells were elicited by stimulating amacrine cell inputs with electrical shocks. Pharmacological analysis indicated that the GABA receptors mediating the IPSCs on these two neurons were different. Synaptic responses in bipolar cells were relatively prolonged and were largely bicuculline or SR-95531 insensitive, indicating that these responses were mediated mainly by GABAC receptors. By contrast, the synaptic responses in ganglion cells have a much more transient time course than the bipolar cell responses and were completely blocked by bicuculline orSR-95531, indicating that they were mediated exclusively by GABAA receptors. Furthermore, current responses evoked by puffing GABA onto bipolar cell terminals or ganglion cell dendrites had time courses and pharmacological profiles that were similar to the synaptically evoked responses. Because puff-evoked responses are not determined by transmitter release kinetics, this result suggests that the properties of postsynaptic GABA receptors can, at least in part, shape the time course of IPSCs in bipolar and ganglion cells.
GABAC receptor ρ-subunit localization studies support our findings. Immunocytochemical studies in mammalian, bird, and fish retinas (Enz et al. 1996; Koulen et al. 1997) have shown that ρ-subunits are localized to bipolar cells but not amacrine or ganglion cells. Furthermore, most of theρ-immunoreactive puncta are localized to the bipolar axon terminals where these cells receive synaptic input from amacrine cells, supporting the notion that GABAC receptors in the IPL are found primarily on bipolar cell axon terminals.
We found that bipolar cell and ganglion cell GABAA receptor–mediated IPSCs had distinct time courses. GABAA receptor–mediated synaptic responses in bipolar cells decayed approximately fourfold more slowly than those in ganglion cells. These differences may reflect different subunit compositions for GABAA receptors on the different cell types. In rat retina, different classes of amacrine cells have particular GABAA receptor subunit compositions, which are distinct from the rod bipolar cell GABAA receptor subunit composition (Greferath et al. 1995). Mammalian cell lines transfected with cDNAs of different GABAA receptor subunits have distinct response time courses (Dominguez-Perrot et al. 1996; Gingrich et al. 1995; Tia et al. 1996). The time courses of these GABA responses depended either on the subtype of α-subunit expressed (Gingrich et al. 1995; Tia et al. 1996) or the presence or absence of an expressedγ-subunit (Dominguez-Perrot et al. 1996; Tia et al. 1996). Additional work is needed, however, to determine whether differences in GABAA receptor subunit composition and/or other factors underlie the differences in GABAA receptor–mediated IPSC time courses.
The time courses of bipolar and ganglion cell responses may also be determined, in part, by differences in the rate of transmitter removal. The distinct patterns of immunolabeling for different subtypes of GABA transporters in the inner retina of rat (Johnson et al. 1996) and salamander (Yang et al. 1997) are consistent with this notion. It is possible that differences in the numbers of transporters or differences in the rates of different transporter subtypes could shape synaptic responses in the salamander retina. However, different GABA transporters probably do not play a major role in shaping responses in our studies. This is because synaptic or puff-evoked responses on the same bipolar cell, which were mediated by GABAA or GABAC receptors, had different time courses. Presumably, the rates of transmitter removal from the two types of GABA receptors were similar for a single bipolar cell. These results suggest that receptor properties were relatively more important than transmitter removal rates in determining synaptic response time courses mediated by GABAA and GABAC receptors.
The time courses of GABAergic synaptic responses in bipolar cells and ganglion cells may also be determined, in part, by different time courses of GABA release from distinct amacrine cells. For example, bipolar cells may receive inputs from sustained GABAergic amacrine cells, and ganglion cells may receive inputs from transient GABAergic amacrine cells. However, sustained, GABAergic amacrine cells have been shown to make synaptic contacts with bipolar cell axon terminals as well as with ganglion cell dendrites in goldfish retina (Muller and Marc 1990). Furthermore, ultrastructural evidence indicates that a single amacrine cell process can make synaptic contacts onto two postsynaptic processes in the tiger salamander IPL (Wong-Riley 1974). This anatomic evidence suggests that, in principal, a single GABAergic amacrine cell process can make synaptic contacts onto both bipolar cell terminals and ganglion cell dendrites. If this is indeed the case, then the time courses of synaptic responses of bipolar and ganglion cells may be determined by the complements of postsynaptic GABA receptors independent of presynaptic differences in transmitter release. We cannot rule out at this time that in addition to GABA receptor properties differences in transmitter release rates might also shape bipolar cell and ganglion cell GABAergic synaptic responses. However, our results clearly demonstrate that the properties of postsynaptic GABA receptors play an important role in shaping the time courses of these responses.
On the basis of the time course of the responses mediated by GABAA receptors, it might be expected that GABAergic responses in ganglion cells are always transient. This may not be the case, however, because sustained GABAergic synaptic responses have been reported in mudpuppy ganglion cells (Belgum et al. 1984). These sustained responses may be due to the temporal and spatial summation of presumed transient GABAA receptor–mediated synaptic responses. Similar findings have been reported for horizontal cells where transient responding α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)–type glutamate receptors may mediate sustained synaptic responses (Eliasof and Jahr 1997). For horizontal cell AMPA receptors, the recovery from desensitization is relatively rapid, allowing for temporal summation. Although our results indicate that synaptic responses mediated by GABAA receptors were relatively brief compared with those mediated by GABAC receptors, it is possible that sustained responses could be mediated by GABAA receptors in ganglion cells, provided that the transient responses can temporally and/or spatially summate. This suggests that neurons that possess GABAA receptors may have finer temporal resolution of inhibitory signals compared with neurons that possess mainly GABAC receptors.
Do ganglion cells have GABAC receptors?
Our results are in agreement with those reported for GABA-evoked currents in mammalian bipolar and ganglion cells (Feigenspan et al. 1993; Lukasiewicz and Wong 1997; Yeh et al. 1996), which showed that ganglion cells had exclusively GABAA receptors and bipolar cells had a combination of GABAC and GABAA receptors. In contrast to these, and our studies, it has been reported that GABAC receptors may mediate a small, but significant (∼20%), component of ganglion cell GABA-elicited responses in tiger salamander (Zhang and Slaughter 1995) and turtle (Liu and Lasater 1994). Messenger RNA for ρ2 and ρ3 subunits has been localized to the ganglion cells in rat retina (Ogurusu et al. 1997; Yeh et al. 1996), suggesting the presence of GABAC receptors on these cells. However, these messages are probably not expressed because only GABAA, and not GABAC, receptor–mediated currents were found in rat ganglion cells (Yeh et al. 1996). Also, the absence of ρ-subunit labeling suggests that these subunits are not expressed in amacrine and ganglion cells (Enz et al. 1996; Koulen et al. 1997). Furthermore, we never saw any evidence of a slowly decaying component of the synaptic response in ganglion cells as would be predicted if a portion of the response was mediated by GABAC receptors.
Salamander bipolar cells may have a different subtype of GABAC receptor
I4AA has previously been shown to act as a GABAC receptor antagonist in fish and rat retinas (Pan and Lipton 1995; Qian and Dowling 1995) and in ρ-subunits expressed in Xenopus oocytes (Kusama et al. 1993). In salamander, however, I4AA acts as a GABAC agonist, suggesting that GABAC receptors on salamander bipolar cells may be different from those previously described in other species. In contrast to the findings that I4AA is an antagonist at native GABAC receptors in white perch retina, Qian et al. (1997) recently reported that I4AA acted as an agonist at ρ-receptor subunits cloned from white perch retina and expressed in Xenopus oocytes. It is possible that a similar ρ-subunit is a component of salamander GABAC receptors.
Roles of GABAA and GABAC receptors for inner retinal processing
Presynaptic GABAC receptors on bipolar cell terminals control the excitatory output from bipolar cells (Lukasiewicz and Werblin 1994). These more sensitive, presynapticGABAC receptors on bipolar cell terminals will be activated at lower synaptic GABA concentrations than the less sensitive GABAA receptors on both postsynaptic ganglion cell processes and bipolar cell terminals. Reciprocal synapses have been reported between bipolar terminals and amacrine cell processes in the salamander retina (Dowling and Werblin 1969; Wong-Riley 1974). Negative feedback could occur at a bipolar cell terminal contacting a GABAergic amacrine cell process. This feedback signal may lead to noise reduction as well as a reduction of synaptic gain at the bipolar cell output as Smith (1995) has proposed for feedback onto photoreceptor terminals. The presence of the more sensitive GABAC receptors on bipolar cell terminals suggests that feedback may occur at lower synaptic GABA concentrations than if only the less sensitive GABAA receptors were present. In addition, the presence of GABAC receptors suggests that feedback will be more prolonged than if only GABAA receptors were present.
The different functional properties of GABAA andGABAC receptors suggest that GABAergic signals in the IPL will have different effects on bipolar and ganglion cells. For example, weak inhibitory surround inputs mediated by amacrine cells may activate more sensitive GABAC receptors on the bipolar cell terminals, but not less sensitive GABAA receptors on ganglion cell dendrites. Previous work has shown that the properties of sustained antagonistic responses in salamander ganglion cells depend on surround intensity (Thibos and Werblin 1987). Dim surrounds cause a parallel rightward shift of the ganglion cell spot intensity-response function. This surround mechanism is consistent with presynaptic inhibition at the bipolar terminals (and/or at photoreceptor terminals) resulting in less transmitter release for a given spot stimulus, i.e., reduced synaptic gain. Brighter surrounds cause a further rightward shift of the spot intensity-response function and a decrease in the function maximum. This is consistent with direct inhibition of ganglion cells causing a decrease in maximal excitation, i.e., a reduced dynamic range. It is possible that the more sensitive GABAC receptors on bipolar cell terminals mediate presynaptic surround mechanisms at dim intensities, whereas the less sensitive GABAA receptors on ganglion cells mediate postsynaptic surround mechanisms at brighter intensities.
We thank Drs. Paul Cook and Rachel Wong for comments on the manuscript.
This work was supported by National Eye Institute Grants EY-08922 to P. D. Lukasiewicz, EY-07057 to C. R. Shields, and EY-02687, a Core Grant to the Department of Ophthalmology, and by the Research to Prevent Blindness.
Address for reprint requests: P. D. Lukasiewicz, Dept. of Ophthalmology and Visual Sciences, Campus Box 8096, Washington University School of Medicine, St. Louis, MO 63110-1093.


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This property has been removed by the agent. It may be sold or temporarily removed from the market
3 bedroom character property for saleRhulen, Builth Wells, Powys
Property Description
Full description
Tenure: Freehold
A stunning three bedroomed stone farmhouse offering high quality living accommodation, two stone barns (one converted, the other with consent) and 2.5 acres of grounds with spectacular views over the Edw Valley.
|6'2" x 4'5" (1.88m x 1.35m). with stone flag floor, windows north west and south east, exposed stone walling, stable door to:
|13'9" x 6'6" (4.2m x 1.98m). (max) with oak flooring and staircase, doors to first floor:
|12'1" x 11'10" (3.68m x 3.6m). with window south west overlooking front driveway and views over the hills , large stone Inglenook fireplace with stone flag hearth and a large Clearview woodburning stove.
|20'8" x 9'6" (6.3m x 2.9m). (max) with windows south west and north east, fitted with a very high quality Keller kitchen with black stone work surfaces with stone splashbacks, one and a half bowl sink, two Bosch ovens, Bosch five burner gas hob, built-in dishwasher, American fridge/freezer.
|22'3" x 18'1" (6.78m x 5.51m). with large double height windows constructed to maximise the views across the stunning Radnorshire hills, further windows north east and south east.
|8'10" x 7'10" (2.7m x 2.39m). with window north east, roll fronted worktop with floor mounted kitchen units, tiled splashbacks, stable door to:
|7'7" x 4'5" (2.31m x 1.35m). with windows north west and south east, black tiled floor, door to exterior.
|8'1" x 2'10" (2.46m x 0.86m). with large feature stone fireplace, half bowl sink unit, spotlighting, low flush WC, MHG Micromatt boiler.
| with window south west, oak flooring, doors to:
|22' x 18'1" (6.7m x 5.51m). (max) with separate dressing room (2.48m x 1.78m) with fitted shelving, balustrade overlooking the double height window with far reaching views over the Radnorshire Hills, revealed roof trusses, door to:
|10'2" x 9'6" (3.1m x 2.9m). (max) with window north east, tiled floor, large double sized shower unit, low flush WC, sink unit, heated towel rail.
|18'1" x 8'2" (5.51m x 2.5m). with windows north east, exposed stone walling, two skylights south east, revealed timber beams, radiator, access door to loft space, shared access to the:
|10'10" x 8'6" (3.3m x 2.6m). with a centrally located bath with tiled surround, low flush WC, heated towel rail, skylight north east, sink unit with mirror over.
|12'4" x 11'4" (3.76m x 3.45m). with window south west, airing cupboard with vertical radiator and access to the above bathroom.
| fully boarded and insulated and containing plant for the Passive House type heating system.
| The property sits in a delightful location and is accessed via the private drive that leads onto a large parking area with ample turning space. To the front of the property is a good sized garden, predominantly laid to lawn and enjoying far reaching views across the Edw Valley. To the rear and side of the property is approximately 2.5 acres of mixed woodland and amenity space and significant habitat areas attracting an abundance of birds and wildlife.
|33'4" x 14'9" (10.16m x 4.5m). A recently refurbished unit providing ancillary accommodation to the house. Currently used as a home office/photographic studio it is finished with antique oak flooring, a partial mezzanine floor accessed via a spiral staircase, revealed roof trusses, vertical radiators, a woodburning stove, French doors south east and north west with beautiful views.
|21'8" (6.6m) x 17'5" (5.3m) plus 19'4" (5.9m) x 17'5" (5.3m). with planning permission for conversion into ancillary accommodation to the property. Currently used as a garage and store room. Predominantly of stone construction with concrete floor.
| The property benefits from a highly efficient and modern heating system of a Passive House type design. Hot water and heating is provided through a system of heat recycling that requires minimal input from existing LPG tanks which ensures that heating bills are minimised. Typical annual hot water, central heating and cooking bills have been £750.
More information from this agent
To view this media, please visit the on-line version of this page at www.rightmove.co.uk/property-for-sale/property-18171927.html
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Disclaimer - Property reference HAY100050. The information displayed about this property comprises a property advertisement. Rightmove.co.uk makes no warranty as to the accuracy or completeness of the advertisement or any linked or associated information, and Rightmove has no control over the content. This property advertisement does not constitute property particulars. The information is provided and maintained by McCartneys LLP, Hay-On-Wye. Please contact the selling agent or developer directly to obtain any information which may be available under the terms of The Energy Performance of Buildings (Certificates and Inspections) (England and Wales) Regulations 2007 or the Home Report if in relation to a residential property in Scotland.
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The other day, I was working on a post about death threats against George Zimmerman. I wanted an appropriate picture to accompany the article. So I did a Google search for images of "blood lust."
Imagine my surprise when this photo of Obama popped up. It is truly the most frightening pic of Obama I've ever seen.
The pic is not a photoshop. It is published on the reputable BusinessInsider.com site, for an article titled "Here's What Obama REALLY Thinks of Wall Street". There's no date for the photo, but it appears to have been taken in September 2009.
Here's the pic:
What creature is in the White House?
God help us.


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You gotta like the Als
All the ingredients appear to be available for the Alouettes to continue their winning ways at Molson Stadium Friday night, against Saskatchewan.
The Als should be stoked after blowing a 21-point lead to the Riders in the opening game, eventually losing in double overtime. And we all know the Als haven't lost at home in more than a full season. Plus, they'll continue pulling away from their East Division rivals with a victory while sending a message to a West Division contender to the throne.
But most of all, Riders' quarterback Darian Durant has been suffering from a stomach virus for close to three weeks. He was held out of practice one day this week, spent the day in hospital, undergoing tests, has lost weight and is likely feeling week while on medication.
Durant's sick. And his teammates will likely be more ill come Friday night.
Als win, 35-16.


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I wouldn't be surprised if these also came in around $17 though.
What can I say? I forget how to read sometimes.
And with Target there is always a better chance of clearances. The Wal-Marts around here NEVER seem to clearance anything.
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BAILEY Rice (Drighlington) was the winner of the David Ike Robinson Memorial Handicap, after defeating Nathan Dobson (Wibsey) 21-15 in the final.
Rice follows in the footsteps of James Davison (Eccleshill), Thomas Hanson (Pudsey), James Hanson (Pudsey), Stuart King (Asa Briggs), Josh Mordue (Brighouse Sports) and Robbie Ellis (Shipley Club) as winners of this prestigious Bradford junior event and will surely be a name to watch in the coming years.
*Wayne Ditchfield (Eccleston) and Hugh Hornby (Preston) were the final qualifiers in the 2014 Spen Masters, sponsored by P & I Hopkins Butchers. Ditchfield defeated Paul Chamberlain (Anglesey) 21-19, Richard Bentley (Cleckheaton Sports) 21-12 and Gavin Parker (Preston) 21-9, drawing number ten on finals night, whilst Hornby drew number five, with wins over David Burns (Peel) 21-14, David Scott (Halifax) 21-17 and Kevin Burns (Ossett) 21-20.
Scores from Week 8 of Qualifying (1st Round): Wayne Ditchfield (Eccleston) 21, Paul Chamberlain (Anglesey) 19; Ashley Daykin (Huddersfield) 14, Richard Bentley (Cleckheaton) 21; Gavin Parker (Preston) 21, Derek Seed (Settle) 15; John Lloyd (Heswall) 21, Roy Nicholson (Halifax) 17; Conor Chamberlain (Anglesey) 5, Kevin Burns (Ossett) 21; Chris Bray (Cleckheaton) 9, Andrew Sykes (Mirfield) 21; David Scott (Halifax) 21, Dan Petcher (Burton on Trent) 20; Hugh Hornby (Preston) 21, David Burns (Bradford) 14; (2nd Round), Ditchfield 21, Bentley 12; Parker 21, Lloyd 14; Burns 21, Sykes 6; Scott 17, Hornby 21; (3rd Round), Ditchfield 21, Parker 9; Burns 20, Hornby 21;
Draw for final (on Saturday, September 6), Simon Coupe (Preston) v Lee Heaton (Chorley), Andy Buckley (Chadderton) v Callum Wraight (Shropshire), Hugh Hornby (Preston) v Scott Woodward (Sheffield), Graeme Wilson (Audenshaw) v Robbie Fitzpatrick (Dewsbury), Kevan Shaw (Bolton) v Wayne Ditchfield (Eccleston), James Hanson (Pudsey) v Thomas Hanson (Pudsey), Phil Lee (Merseyside) v Ashley Tattersley (Huddersfield), Chris Kelly (Sheffield) v Graham Hickey (Halifax);
*The Yorkshire Cup final takes place on Sunday at Almondbury Libs, with the 2012 winners Thongsbridge, taking on the 2013 winners Lower Hopton. Both teams are full of quality, with many tipping Lower Hopton as pre-match favourites. Draw for final (Lower Hopton names first), Paul Ingleby v Richard Jackson, Robert Hitchen v Andy Farmer, Craig Newton v Chris Kelly, Graeme Wilson v Ryan Clark, Harry Seehra v Andy Cairns, Ashley Tattersley v Andy Spragg, Chris Brown v Chris Hill, Ian Booth v Craig Hill, Steve Pinfield v Mark Booth, Simon Walder v Jack Clarke, Graham Higgins v James Davison, Graham Hickey v Andrew Sorsby.
*Tony Riley won the Ben Ramsden Merit, for the seventh time, after defeating Mark Dobson (Wibsey) 21-15 in the final. Riley first won the Ben Ramsden in 1981, and is still classed as one of Bradford's finest bowlers. Full report will appear in next week's T&A.
*Entries are still been taken for the Walter Worsman Teenage Handicap, sponsored by Alpha Graphic, which takes place at Birkenshaw on Sunday 7th September. Anyone interested should contact Norman Lickley on 01943-884323.
*The Concept League is in its sixth season and is run and organised by David Beardsworth. The Concept League currently has 12 teams in it, mainly from the Aire Wharfe area, but David is looking to expand it to sixteen teams for the 2015 Season. It is played on Tuesday evenings, with teams consisting of eight players, using the 54321 scoring system. Anyone interested should contact David Beardsworth on 0113-2573086.


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NFL Draft 2014: Instant-Impact Prospects Who Will Slide out of Round 1February 24, 2014
The 2014 NFL draft features one of the deepest classes in years and there are plenty of exciting prospects to be found after the first round.
According to Alan Robinson of the Pittsburgh Tribune-Review, Pittsburgh Steelers general manager Kevin Colbert recently said it was the deepest class in 30 years, though he mentioned that some of the underclassmen should have stayed in school for more seasoning:
With so many talented players to chose from, there are bound to be at least a few instant-impact prospects who will fall through Round 1.
Not only will they be tremendous steals for their new teams, but they'll likely play with a huge chip on their shoulders after being passed over in the first place.
Carlos Hyde, RB, Ohio State
It will be completely shocking if a single running back is selected in Round 1 this year. The position has seen its value decline over the past couple of years as more and more excellent players are being found in the latter rounds.
"It does kind of bother me," Hyde told reporters at the combine. "I feel like they are just down on us. They don't think we are capable of doing what we know we can do. They are kind of just downplaying us: 'We can just wait to get ya'll' (later in the draft)."
If any back is going to sneak into Round 1, it's Carlos Hyde out of Ohio State. Unfortunately, Scott Wright of DraftCountdown.com reports that he hurt his hamstring running the 40-yard dash at the combine:
A team in need of a new starting running back would be smart to target Hyde in Round 2, however. He's ready to step right into the starting lineup and contribute in a big way, featuring plenty of size, agility and power to go along with adequate speed.
Hyde will have an Eddie Lacy-type impact for his new team in 2014 be an early favorite to win the NFL's Offensive Rookie of the Year award.
Kyle Van Noy, OLB, BYU
This year's draft features many talented pass-rushers, with Jadeveon Clowney out of South Carolina, Khalil Mack out of Buffalo and Anthony Barr out of UCLA leading the class.
Kyle Van Noy has somewhat fallen through the cracks after a relatively quiet season during which he logged just four sacks, but he's primed to have a huge impact at the next level.
In his career at BYU, Van Noy tallied 26 sacks, seven interceptions, 11 forced fumbles and scored five touchdowns, showing excellent playmaking abilitiy.
According to Mike Wobschall of Vikings.com, he said, "I feel like I'm an offensive player on defense."
While Van Noy didn't wow on the track at the combine (4.73-second 40-yard dash), he's a natural athlete with excellent instincts and a knack for making plays on the ball. Even better, he's capable of thriving in any scheme, though he'd be best-suited as an outside linebacker in a 3-4.
Troy Niklas, TE, Notre Dame
Every team would love to have Rob Gronkowski on its roster, which is why Notre Dame's Troy Niklas will likely be scooped up sometime in Round 2 despite the fact he didn't make a huge impact during his tenure at Notre Dame.
A former starter as a true freshman outside linebacker on defense, Niklas was moved to tight end by head coach Brian Kelly before the 2012 season.
In two seasons, he caught just 37 passes for 573 yards and six touchdowns, but don't let the lack of production fool you. Niklas is a star in the making.
Measuring in at 6'6" and 270 pounds, he's a phenomenal run-blocker who understands how to gain leverage against bigger defensive players, as pointed out by Bleacher Report's Matt Miller:
When asked to kick out a defensive end or linebacker in the run game, Niklas puts all 270 pounds into his blocks.
I talk a lot about three-down tight ends, and Niklas is pro-ready right now.
In addition to his run-blocking prowess, Niklas is an excellent receiver who uses his body well to shield defenders. He'll become an integral part of the offense in short order for the team that drafts him this year.
All combine results courtesy of NFL.com's results tracker.
Follow me on Twitter @JesseReed78


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Ryne Sandberg's NL East Preview
Baseball Hall of Famer elect Ryne Sandberg's NL East preview was posted on Yahoo Sports yesterday. Sandberg picked the Mets for second place this year but says they could be the team to end Atlanta's reign over the division. I'm not quite that confident that the can be that good but I know they'll be making some noise this season.
If you enjoyed this post, please consider to leave a comment or subscribe to the feed and get future articles delivered to your feed reader.


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International Racing News
Magnificent March Builds To $10m Finale
Mar 8 2012
The World Cup caps a remarkable month of international racing with major events from Hong Kong, South Africa, New Zealand and the great Cheltenham Festival. The world's richest race has captured international attention from its first running, when the great Cigar claimed the pirze. It's since been won by champions Singspiel and Dubai Millennium and the great American Curlin took the event at the height of his powers in 2008.
If So You Think can win this year's renewal, he will confirm his greatness and silence some of the knockers who were somehow disappointed with three Group 1 wins and two seconds during an arduous seven-race preparation that took him from Ireland to England, France and the United States.
The dual Cox Plate winner is ideally suited in Dubai World Cup but early indications are that he will face strong opposition from all around the globe. The list includes Kentucky Derby winner Animal Kingdom, Japanese champions Transcend and Smart Falcon, Singapore Cup winner Gitano Hernando, US star Game On Dude, South African champion Bold Silvano and class French galloper Byword.
On class and overall form, So You Think will be the horse to beat. He is the best performed horse in the race – even allowing for the fact Animal Kingdom was brilliant in last year's Kentucky Derby – and the 2000m is his ideal distance.
The Dubai World Cup is shaping as a truly great international race this year but it's no stand-alone event – the undercard features Group 1 races the Duty Free, Sheema Classic, Kahayla Classic, Golden Shaheen – all of which should attract world-class gallopers.
Shane McNally
<<< Back to International News


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Questions raised about Farm Bureau endorsement of Sasse
Posted: 07/30/14 08:29
Nonpartisan U.S. Senate candidate Jim Jenkins, sent a letter to the Nebraska Farm Bureau yesterday questioning their endorsement of Ben Sasse in the U.S. Senate race to replace retiring Senator Mike Johanns. In the letter Jenkins writes
"While I realize that the Nebraska Farm Bureau decided to endorse Mr. Sasse early on, I would like to place my rural resume in front of you and allow you to compare it to that of Mr. Sasse who, while very intellectual, has virtually no agricultural or private sector experience and a very thin Nebraska resume, having moved back to the state only five years ago."
That letter sparked a response from Steve Nelson the President of the Nebraska Farm Bureau who said "Our next U.S. Senator will face many challenges. One of the largest is tackling the issue of runaway federal spending and government growth. The future prosperity of Nebraska farm and ranch families is dependent on Congress' ability to get our country's fiscal house in order, which has a direct connection to the development of sound farm policy. We believe Ben Sasse is the right person to address this critical issue and with that ensure, we continue to have federal farm policy that meets the needs of farmers, ranchers and all Americans who benefit from the food, fuel and fiber derived from agriculture."
Republican United States Senate candidate Ben Sasse says he understands why independent candidates might have an appeal for voters this fall.
Two independents have joined the November ballot with Sasse and Democrat Dave Domina. Sasse says voters have seen enough of the two parties which dominate Washington. Both Jim Jenkins of Callaway and Todd Watson of Lincoln have qualified for the ballot.
As per Federal Communications Commission Regulations following is certified: Platte River Radio Inc. does not discriminate in the sale of commercial time, and will not accept advertising which, in its sole opinion, is purchased with intent to discriminate unlawfully on the basis of race, gender, or ethnicity. The advertiser hereby certifies that its purchase of commercial time is not made for an unlawful discriminatory purpose, including specifically that it is not based upon a decision to place advertising on a station on the basis of race, gender, or ethnicity.


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The placard had been left at Hovde Hall on Monday by protesters upset over the recent discovery of the words "white supremacy" written on a mirror inside the campus' Black Cultural Center.
"Police are investigating the incident as a hate crime since property was altered or destroyed and the conduct was apparently motivated by bias," the university said in a news release.
The office of Dean of Students Danita Brown also was investigating the incident as a violation of the student code of conduct that could result in expulsion, the news release said.
Purdue President Mitch Daniels issued a statement saying "bigotry and hate are neither welcomed nor tolerated at Purdue University."
"There is no place at Purdue for those who act from hatred. If you recognize yourself in this last sentence, you are not welcome at Purdue," Daniels said in an apparent response to a demand by Purdue's Anti-Racism Coalition that he should "articulate a zero-tolerance stance against all racist acts."
More than 200 black, white, Asian and Hispanic students, faculty and staff marched Monday to the steps of Hovde Hall chanting slogans such as "This is what diversity looks like!" and "The people are the power!"
Purdue has said in a statement that the "white supremacy" inscription was not an act of vandalism but was an unintentional transfer of words from a sticky note during an educational seminar.
Nonetheless, organizers of the protest said the incident, whether intentional or not, followed a series of race-related incidents on campus.
FBI statistics rank Purdue second in the nation among public and private universities for the number of reported hate crimes. Those rankings are not objective, however, because reporting of hate crimes by universities is inconsistent.
The demands shouted Monday by the Anti-Racism Coalition also call for doubling the number of minority faculty and students over the next 10 years and requiring an undergraduate course on race and racism.
"All diversity means is difference, unlikeness," Christopher Warren, who instructs courses on African-American studies and sociology at Purdue, said during the protest. "Who cares about diversity when there is no equality?"
Provost Tim Sands watched the demonstration from the sidelines and walked up to the first step of Hovde Hall and addressed the gathering when the crowd called his name.
"We are not an inclusive environment," said Sands. "We have not figured that out yet ... I'm really excited to see this momentum building. I'm sorry it's based on the fact that Purdue is not yet a psychologically safe place to study for many people. It is for some but not for everyone."


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Showing Suppliers 1 - 2 of 2
Linx Technologies (38 Products)
Address: 159 Ort Lane, Merlin, OR
Business Type: Manufacturer
Description: An Internet of Things Products and Services Provider At its core, Linx Technologies is an RF module company. We are world leaders in supplying radio modules to remote control applications and we have extremely competitive modules for data transfer. Our tag line is Wireless Made... (more)
Future Electronics
Address: 6650 SW Redwood Lane, Suite 365, Portland, OR
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Merrill Lynch has agreed in principle with US securities regulators to pay $80m (£50m) to settle an investigation into transactions it arranged for failed energy trader Enron.
The Merrill agreement which is still to be finalised by the Securities and Exchange Commission would represent the first civil penalties federal regulators have extracted from the many Wall Street investment banks that were involved with Enron.
Merrill did not admit or deny wrongdoing.
The agreement, outlined in a statement last night by Merrill, is likely to prove a benchmark for other banks, including Citigroup and JP Morgan Chase, which earned hundreds of millions of dollars in fees from the company and have subsequently come under scrutiny since its collapse.
Inquiries into Merrill's involvement with Enron have centred on a series of complex transactions involving electricity-generating barges in Nigeria in 1999.
A Senate sub-committee alleged last year that Merrill had agreed to buy the barges from Enron only after assurances that the energy group would repurchase them within six months at a specified premium.
The transaction, the committee alleged, was concocted as a sham to inflate Enron's year-end earnings.
Merrill denied this at the hearing. Two months later, it fired a vice chairman, Thomas Davis, and its top energy banker, Schuyler Tilney, for refusing to testify to the SEC and Justice Department about the transactions.
Merrill's settlement is expected to shift the regulatory focus to Citigroup and JP Morgan and their dealings with Enron. Both banks were accused in Congressional hearings of camouflaging loans to Enron as commodities trades to help the company conceal its true debt levels.
Merrill said it would account for the settlement as part of last year's fourth-quarter results. Last month Merrill said it had made a net profit of $603m during the period. However, it was unclear how much earnings will be reduced because the bank has yet to decide on the tax deductibility of the settlement costs.
The charge comes in addition to the $200m that Merrill has agreed to pay to settle investigations into conflicts of interest in its equity research and public offering procedures.


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Are you sick of Johnny Manziel yet?
We sure hope not, because CBS will be devoting an entire camera during its college football coverage on Saturday to follow the reigning Heisman Trophy winner around during Texas A&M's matchup against Alabama, SI.com reports.
The camera will be called the "Johnny Cam."
"No matter where he is and no matter what part of the game it is, we will have a shot of it," said Craig Silver, the coordinating producer of college football for CBS Sports. "If he is anywhere in sight of that camera, we will catch it."
The game is one of the most anticipated of the college football season, with fans looking to see whether or not Manziel lives up to his hype and Alabama looking for a three-peat in the national championship department. Manziel's off-field antics have been covered ad nauseam this past the offseason, but Silver said viewers shouldn't expect to hear all Manziel all the time on Saturday.
"By the time we get to kickoff, this thing will have been talked about ad nauseam throughout the sports world," Silver said. "We are not judge and jury. It is not our place, especially within the body of the broadcast, to state whether he should or should not be playing, should have been suspended or should not have been suspended. The way I approach it is how has all this stuff affected him as a football player and affected his team."
CBS requested an on-camera interview with the quarterback, as well as his presence for meetings with the CBS production staff this Friday. Texas A&M will make decisions based on those requests this week.
ESPN's College GameDay will also be on hand in College Station, Tex., to broadcast its college football show. Executives echoed Silver's sentiments about the oversaturation of Manziel-related news.
"I can promise you the show won't turn into 'Manziel Mania' for three hours," ESPN coordinating producer Lee Fitting said. "Will the viewer get their fill of Manziel? Sure. Will we cover every angle of the game? Yes. But will we go overboard? Absolutely not."
Sure, ESPN. We believe you.
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Store Bought Rkt Falling Apart! Leahs Please Help
Decorating By Rocketgirl899 Updated 7 Dec 2008 , 3:32am by Rocketgirl899
okay, so I am babysiting (don't worry kids are now in bed) and my dad calls me to tell me my bowling pin (12-16inchjes) is starting to fall apart. Instead of making the RKT I bought it premade from Sam's club ( i heard some other ccers just do this) In the past I have made RKT and left out almost all the butter and didn't seem to have a problem.
Well i am watching the kids till 11pm EST and have no idea what to do. the cake is for my birthday party tomorrow at 3.. but everyone is expecting something amazing. (The cake will actually be a bowling ball, its a small party)
Is there something I can do to make them stiffer? or should I just go to the store and make my own? I would have made my own, but I had 2 papers (20 and 15 pages) plus a video for school this week as it is finals.
any thoughts would be great, I am going to ice it in BC.
Do you think I need a support dowel in the middle? what size?
I used a 2liter bottle for the initial shape and then added hieght to it.
thanks for the help. I feel so helpless right now. Why did i ever agree to babysitt this week! lol
I know Leah, you said you use the store bought, maybe I didn't compress them enough?
I think if i don't figure something out by the time i go home, im just goign to wrap it in duck tape and make it non edible . I still haven't started to fill or ice the balls, did i mention I have never done this kind of cake? lol.
How about melted chocolate? When it hardens up it's pretty firm. You could "glue" the pieces together and wrap them in saran wrap to hold the form until it cools, then unwrap and ice.
thats a pretty good idea.. i think i MIGHT have like 1 cup of candy melts? ergh. thanks
other than that i know I dont havge any other chocolate..
thats better than my duck tape idea tho... lol. did i mention my brain is fried from finals and im workin with 3 hours of sleep in the past 3 days
I do use the premade ones, and I squish the He// out of them. They are smooth on the outside they are so compressed. So candy clay or fondant on the outside should hold things together, though.
I use store bought RK treats all the time and yes they are not as secure as homemade. I squish them together as much as I can so there is no air. Once I have formed what I need, I paint it with thinned RI. I haven't tried chocolate, but I found that RI works great for me. I usually put on a few coats. And I always cover it in fondant for a clean look. But I have coverd the whole thing in RI as well (i.e: Elmo) and it dried rock hard and was fine.
Good luck. I feel for you.
well, I got home, and my awesome (well for what I can do ) bowling pin is a pile of RKT.
I will sqqeeze the bigeebees outta them and try that. I thought I had them pretty compact. I guess not. I also found about a cup of candy melts I will try to use. Thanks.
ended up compacting more..... and it worked great. I had to make some RKT cause I ran out and i put them in the food processor and left out the butter. Worked great hard as a rock. but i had a tough time shapping it after all was said and done so im not happy with the overall shape.
MANY problems with this bowling pin.
too embarrassed to post photos
thanks for the help


text: +-------------------------------------------------------------------------+ | TPWD News Release 20120312a | +-------------------------------------------------------------------------+ | This page contains only plain text, no HTML formatting codes. | | It is not designed for display in a browser but for copying | | and editing in whatever software you use to lay out pages. | | To copy the text into an editing program: | | --Display this page in your browser. | | --Select all. | | --Copy. | | --Paste in a document in your editing program. | | If you have any suggestions for improving these pages, send | | an e-mail to firstname.lastname@example.com and mention Plain Text Pages. | +-------------------------------------------------------------------------+ [ Note: This item is more than five years old. Please take the publication date into consideration for any date references. ] [ Media Contact: TPWD News, email@example.com, 512-389-8030 ] [ Additional Contacts: TPWD Wildlife District Leader Calvin Richardson in Canyon, (806) 651-3014, firstname.lastname@example.com; or TPWD Wildlife Diversity Biologist Sean Kyle in Lubbock, (806) 742-4735, email@example.com ] March 12, 2012 Lesser Prairie-Chicken Aerial Survey Begins LUBBOCK - This spring, Texas Parks and Wildlife Department will collaborate with state fish and wildlife agencies from New Mexico, Oklahoma, Kansas and Colorado, West Ecosystems, Inc., and Texas Tech University to conduct a large-scale aerial survey for lesser prairie-chicken booming grounds across the High Plains region in all five states. Information from these surveys will be used by TPWD and the other state agencies to help conserve the bird in partnership with landowners and private industries, including oil and gas, wind energy, and electric utilities. "These surveys represent a unique effort by state fish and wildlife agencies across the region to work together to monitor population trends over time and to effectively manage for this species," said Lubbock-based TPWD wildlife diversity biologist Sean Kyle. "This information, combined with other ongoing conservation efforts, could help avoid the need to list the lesser prairie-chicken as federally endangered. The surveys will provide more data to inform the US Fish and Wildlife Service's decision about whether to list the species." Surveys will begin about March 24 and will continue through mid-May. Agency staff from the five states have randomly selected about 200 eighty-five square-mile blocks located in and around the estimated range of the species. These blocks cover an area ranging from the southern plains of the Texas Panhandle and southeast New Mexico up to northern Kansas and Colorado. Surveys will be flown at 35-40 mph about 80 feet above ground. Pilots will fly two to sixteen north-south oriented transects across selected blocks. Towns, feedlots, and houses will be avoided. Pilots involved have extensive experience conducting aerial surveys and gathering livestock with helicopters. They will make special effort to avoid hazing livestock on pastures. -30-


text: Kaye Whitney, the new pastor at Point Reyes Presbyterian Church, felt a calling to the ministry over four decades ago, but gender norms dissuaded her. "Back when I was a young girl, women weren't necessarily accepted into the ministry… So while it called to me, I didn't see a way to enter it." Instead she became a math teacher. Though it was a field for which she also felt suited, the call to the church remained, and a few years ago she saw a newspaper ad for a seminary program and decided to go for it. "This is your chance, kid," she told herself. When she enrolled at the San Francisco Theological Seminary at 61, she was one of the oldest students. Ms. Whitney was hired as an interim pastor at Point Reyes Presbyterian after the retirement of Carolyn Osborne, and her position was made permanent last month. It is her first job as a pastor. Ms. Whitney, who lives in Benicia and also volunteers as a hospital chaplain in Antioch, is still getting to know the community. "We want people to know we're welcoming," she said. The congregation is small, about 15 to 20 people, but asked whether that's something she might try to change, she said, "That's something God grows."


text:
About 50 anti-war protesters clad in pink made their way onto the Golden Gate Bridge flashing peace signs, waving banners and slowing traffic Thursday morning.
The anti-war group Code Pink staged the protest as part of International Peace Day.
The group applied for a permit to march on the bridge between 10 a.m. and noon, outside of commute hours, bridge officials said.
But the group arrived by 7:30 a.m. on both sides of the bridge and began walking toward the center of the span on the east sidewalk.
By about 8:50 a.m., they met at midspan, pulled out banners and began waving them, which they agreed not to as part of the permit arrangement, officials said.
The group also waved at drivers, which slowed traffic on northbound approaches. Bridge officials opened a third lane going northbound to ease traffic.
The protesters left the span about 10 a.m., officials said.
- Mark Prado


text:
Family Law
Family Law Questions? Ask a Family Lawyer Online.
I would like to add the my x is in the air force and they have their own set of rules. Also, the money that he suppressed was mostly BAH at dependent rate. So the funds that he hid was money the military pays to him because he has children to support.
The child support order that the income was suppressed on happened 3 1/2 years ago. I called child support and told them that something didn't seem correct however my case worker didn't even respond. I had the case review this last month and my x suppressed his income again. I am now fighting that review. Is their any legal option that I have to recover the lost support (over 8K) or am I'm simply out of luck. If their is anything that I can do to collect this support please let me know.


text:
Effects of rhIGF-1 on Bone Metabolism in Adolescent Girls With Anorexia Nervosa (814)
This study has been completed.
Massachusetts General Hospital
Information provided by (Responsible Party):
Madhusmita Misra, Massachusetts General Hospital
ClinicalTrials.gov Identifier:
First received: July 18, 2008
Last updated: February 28, 2012
Last verified: February 2012
- Full Text View
- Tabular View
- Study Results
- Disclaimer
- How to Read a Study Record
Adolescents and young adults with anorexia nervosa (AN) are at high risk for low bone mineral density at a time when healthy adolescents are rapidly accruing bone, with implications for peak bone mass and fracture risk in later life. They are also deficient in insulin-like growth factor 1 (IGF-1), the bone trophic factor made in the liver in response to growth hormone (GH), despite elevated levels of growth hormone. It is possible that deficiency of insulin-like growth factor 1, a hormone very important for the maintenance of skeletal integrity, may contribute to the severe osteopenia seen in anorexia nervosa. The physiologic effects of recombinant human insulin-like growth factor 1 (rhIGF-1) treatment in adolescents and young adults with anorexia nervosa have not been studied. The goal of this proposal is to investigate the effects of recombinant human insulin-like growth factor 1 on bone density and bone microarchitecture in adolescent girls and young adult women with anorexia nervosa over a 6 month period. We hypothesize that adolescent and young adult anorexia nervosa patients, being insulin-like growth factor 1 deficient, will respond to exogenously administered recombinant human insulin-like growth factor 1 with elevations in biochemical indices of bone turnover and an increase in bone density and improvement in bone structure, or maintain bone density (in contrast to the decrease in bone density expected in adolescent girls and women with anorexia nervosa who are not treated).
|Disorder of Bone Density and Structure, Unspecified
|Drug: Recombinant human insulin like growth factor-1 (rhIGF-1)
|Phase 2
|Study Type:
|Study Design:
|Intervention Model: Single Group Assignment
Masking: None (Open Label)
Primary Purpose: Treatment
|Official Title:
|Effects of Recombinant Human Insulin Like Growth Factor-1 (rhIGF-1) on Bone Metabolism in Adolescent Girls With Anorexia Nervosa
Resource links provided by NLM:
MedlinePlus related topics: Bone Density
Drug Information available for: Insulin Insulin-like growth factor I Mecasermin Mecasermin rinfabateU.S. FDA Resources
Further study details as provided by Madhusmita Misra, Massachusetts General Hospital:
Primary Outcome Measures:
- Change in Spine Bone Density (g/cm^2) [ Time Frame: Baseline and 6 months ]Change in spine bone density over 6 months (6month data- baseline data). Bone density at the spine was assessed using dual energy x-ray absorptiometry at baseline and 6 months and the change in bone density over these 6 months was calculated.
|Study Start Date:
|July 2008
|Study Completion Date:
|October 2011
|Primary Completion Date:
|October 2011 (Final data collection date for primary outcome measure)
|Assigned Interventions
|Experimental: Anorexia Nervosa Females
Drug: Recombinant human insulin like growth factor-1 (rhIGF-1)
Twice daily
Other Name: Tercica
|Ages Eligible for Study:
|12 Years to 26 Years (Child, Adult)
|Sexes Eligible for Study:
|Accepts Healthy Volunteers:
Inclusion Criteria:
- Meet criteria for anorexia nervosa (AN) as described in the Diagnostic and Statistical Manual of Mental Disorders
Exclusion Criteria:
- Pregnant or nursing
- Hematocrit < 30%, K < 3 mmol/L
- Any illness (other than AN) known to affect bone and mineral metabolism such as diabetes, untreated hypo- or hyperthyroidism, or hyperparathyroidism
- History of use of any medicine, such as corticosteroids, known to affect bone density. Subjects who are on estrogen will still be eligible to participate in the study because our data in adult women with AN do not indicate deleterious effects of estrogen in patients receiving rhIGF-
Contacts and Locations
Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the Contacts provided below. For general information, see Learn About Clinical Studies.
Please refer to this study by its ClinicalTrials.gov identifier: NCT00720122
Please refer to this study by its ClinicalTrials.gov identifier: NCT00720122
|United States, Massachusetts
|Massachusetts General Hospital
|Boston, Massachusetts, United States, 02114
Sponsors and Collaborators
Massachusetts General Hospital
|Principal Investigator:
|Madhusmita Misra, MD
More Information
Keywords provided by Madhusmita Misra, Massachusetts General Hospital:
Additional relevant MeSH terms:
ClinicalTrials.gov processed this record on September 21, 2017
|Responsible Party:
|Madhusmita Misra, Asssociate Professor of Pediatrics, Massachusetts General Hospital
|ClinicalTrials.gov Identifier:
|NCT00720122 History of Changes
|Other Study ID Numbers:
|Study First Received:
|July 18, 2008
|Results First Received:
|January 27, 2012
|Last Updated:
|February 28, 2012
Keywords provided by Madhusmita Misra, Massachusetts General Hospital:
Bone density
Bone turnover markers
Bone structure
Anorexia nervosa
Additional relevant MeSH terms:
Anorexia Nervosa
Bone Diseases
Signs and Symptoms, Digestive
Signs and Symptoms
Feeding and Eating Disorders
Mental Disorders
Musculoskeletal Diseases
Insulin, Globin Zinc
Hypoglycemic Agents
Physiological Effects of Drugs
Mitosis Modulators
Molecular Mechanisms of Pharmacological Action
ClinicalTrials.gov processed this record on September 21, 2017


text:
With an intense desire I poured out my heart to her.
San Diego (PRWEB) March 27, 2014
The author known as P. A. Ramour reflects on his journey through life with humor, passion and nostalgia in his new poetry collection, "Murmurings of the Heart: One Angelic Kiss" (published by iUniverse).
"My poems were inspired by the beauty and talent of a new found love in my Golden years that I call 'Angel' and I was compelled by her 'One Angelic Kiss' to express my love for her in the ardent form of verse," Ramour writes. "With an intense desire I poured out my heart to her."
Expounding on themes of friendship, unrequited love and appreciation of nature, "Murmurings of the Heart" takes readers through the author's childhood, friendships and life in northern Minnesota, all the way through his golden years. His words offer insight, encouragement and a testament to the power of faith and hope.
An excerpt from the poem, "Companionship":
"Though from a different harbor in a distant nation,
we'll come together in our common destination.
She's like an enchantress dancing
with silken gowns flowing.
Our companion ship has my heart glowing
here on the briny."
"Murmurings of the Heart"
By P. A. Ramour
Hardcover | 6 x 9 in | 118 pages | ISBN 9781475957945
Softcover | 6 x 9 in | 118 pages | ISBN 9781475957921
E-Book | ISBN 9781475957938
Available at Amazon and Barnes & Noble
About the Author
P. A. Ramour grew up one of seven children and learned the carpentry trade after high school, becoming a master carpenter. He was widowed after 35 years of marriage and raising five children and met his "angel of love" while entertaining at a retirement community for seniors. He currently lives in California.
iUniverse, an Author Solutions, LLC, self-publishing imprint, is the leading book marketing, editorial services, and supported self-publishing provider. iUniverse has a strategic alliance with Indigo Books & Music, Inc. in Canada, and titles accepted into the iUniverse Rising Star program are featured in a special collection on BarnesandNoble.com. iUniverse recognizes excellence in book publishing through the Star, Reader's Choice, Rising Star and Editor's Choice designations—self-publishing's only such awards program. Headquartered in Bloomington, Indiana, iUniverse also operates offices in Indianapolis. For more information or to publish a book, please visit iuniverse.com or call 1-800-AUTHORS. For the latest, follow @iuniversebooks on Twitter.


text:
When it comes to product and services selection, buyers today are much better informed than they were 10 years ago. Social networks, peer groups, company websites and industry analysts all arm prospects with more than enough data to help them through the research and evaluation phases of the buy cycle. As they enter the formal phase of the buy cycle buyers usually know the problem they are trying to solve, the solutions available to solve the problem, and a good idea of the cost of the solution.
This means that when your sales team engages with a qualified prospect they may be well into the research and decision phase of the project.
If you own the marketing function this trend means that identifying and doing initial qualification is no longer enough. You must find ways to engage with the prospect earlier in the buy cycle and find ways to out-educate your competition while the hidden sales cycle is underway. Content-based marketing is what best in class companies are investing in to ensure they are engaging with buyers during the hidden part of the sales cycle. It's no longer about selling it's about educating.
In a recent Aberdeen Group report 58% of marketing executives indicated they were improving marketing content creation and only 6% indicated that marketing content was not on their agenda. This trend also shows how critical communication and alignment with sales has become. Developing meaningful content is time consuming and many companies fail in producing the right materials because they don't fully understand the commitment in terms of time and money. The same report suggested that best-in-class companies have well articulated sales processes that have been agreed to between the groups. Additionally, 86% of best in class companies said that the relationship between sales and marketing was either good or strong!
Consider implementing a content-based marketing program so you are not disadvantaged when buyers enter the formal phases of the buy cycle. Our next post will dive more deeply into content marketing best practices and the results you can expect from the investment.


text:
A few hours ago, Derpy went back up for sale over at Hot Topic! It looks like the fixed bubble version from the image, and I don't see any other changes. Chances are its going to disappear in a few hours after this goes up, so be sure to grab one if you want one (assuming it follows the trends of the last few times it went up in stock).
Find it here!
Update: The less rare, but also hard to find Rainbow Dash is also available. Find it here!
Thanks to Bl8demast3r for the heads upSaturday, December 29, 2012 op 5:44 PM
Labels: Merchandise, News, Not-Fanfiction
For archival purposes, you can find the IntenseDebate comments for this post (if any) archived over here
Derpy and Rainbow Back in Stock at Hot Topic


text:
Should Your HOA Manage Itself or Hire a Professional Manager?
Does your community need a professional HOA manager or can volunteers handle the load? Read
Delinquent Home Owners Association Fees: Why You Must Collect
Let your neighbors slide on paying their home owners association fees, and other owners have to pick up the slack. If it gets too bad, home values and sales will fall. Read
No Mothers-In-Law Allowed: The 7 Craziest Local Laws and HOA Rules
Get up to speed with the most outrageous laws and homeowners association rules. Read
Hiring an HOA Manager: What to Consider
When your homeowners association has more to do than its volunteers can handle, it's time to call in a professional manager. Here are 10 tips for picking the right one. Read
Condo Association Newsletter Helps Owners Feel More at Home
Creating a newsletter with all the neighborhood news informs home owners and helps them become part of the family. Read
Visit houselogic.com for more articles like this.
Copyright 2011 NATIONAL ASSOCIATION OF REALTORS®


text:
|Sport, 1 pitch, 65'
|YDS: 5.10a French: 6a Ewbanks: 18 UIAA: VI+ ZA: 18 British: E1 5a [details]
|Jason Desmond
|New Route:
|Page Views:
|Submitted By:
|Jason Desmond on Nov 9, 2010
|Located in a National Forest Fee Area MORE INFO >>>
|Comments on Emotional Geometry
|Add Comment
|Show which comments —
By Ken Noyce
From: Layton, UT
Dec 9, 2010
|Nice job, I've wanted to bolt that route for years. I can't wait to get on it.
By KipHenrie
From: Farmington, utah
Jun 1, 2011
rating: 5.10b/c 6b 20 VII 20 E2 5b
|The name for this is really good. The upper crack area is emotional and its an unusual crack feature for the area with its bizarre geometry. Off width crack with all kinds of different holds and body positioning. The trickiest part being the feet. Have fun but be careful, its still getting cleaned up. We had some rock fall doing it.
By Ryan Stott
From: Salt Lake City, UT
Jun 16, 2011
|VERY CHOSSY. As mentioned, this route has some fun, interesting crack/layback movement that you dont find in AF very often. I'd rate this climb higher if it didn't have so much loose rock. Wear a helmet on belay if you have it.
By Aaron Shields
From: Lehi, UT
May 21, 2013
rating: 5.10b 6a+ 19 VII- 19 E2 5b
|May 2013 - pretty clean now, though the crack is still painfully rough. Good thing it's a short crack! Awesome natural continuation of stoic calculus, and not bad if your crack technique is good
By Jeremy Noring
From: Salt Lake City, Utah
Mar 25, 2014
rating: 5.10d 6b+ 21 VII+ 21 E3 5b
|I'm not sure if something changed, but this route felt significantly harder than 10a or 10b to me. Possible something broke? I'm no crack climber, but the 11b/c around the corner felt a lot easier to me...
By Matt Olson
From: Salt Lake City
Oct 29, 2014
rating: 5.10b/c 6b 20 VII 20 E2 5b
|Great route! Definitely worth jumping on. Seems like there is only a small portion that requires actual crack climbing technique. I'm not a crack climber. Bolts are placed well and found no choss. There's a great knee-bar halfway up the crack that saved me.
By jeff p
From: Golden
Mar 13, 2015
rating: 5.10a/b 6a+ 19 VI+ 19 E2 5b PG13
|Very unique to have bolts on a nice crack, but I'm glad I got to jump on it. My partner and I used it as a good route to practice multi pitch, but it is definitely doable from the ground with a 70 meter rope and a crap ton of draws. Belay station at the first set of anchors is comfy and you can easy see/hear your climber. I did find some loose rocks in the crack but nothing I would say is cause for concern,,, just have your belayer wear a helmet.
By CHopwood
From: Salt Lake City, UT
Mar 21, 2016
|+1 run this as one 35m pitch. So nice to do a long pitch in this canyon of short routes. And how often do you get to do a sport climb and finish on a fun hand/offwidth crack?! Warning, the crack is super sharp so prepare to lose some skin if you jam too deep into it.
By samwisesam Treharne
Oct 25, 2016
|If you want to lead stoic calculus, then straight into emotional geometry, you will need 16 draws. Use only one at the chains of stoic calculus and you will have enough.
By grog m aka Greg McKee
Jun 30, 2017
|I am a crack whore so I was PSYCHED to see this. I climbed it from the ground up, super fun pitch and I highly recommend doing it as one long pitch. Crack is very clean, no choss. There are no offwidth moves unless you want to get crazy and make this way harder than necessary. 5.10a hand jamz.


text:
WellSpring of Faith and Hope Community School kids with new school supplies
WellSpring of Faith and Hope Community School kids receive new shoes
Girls in Sarah's weekly Bible-study class get pillowcase dresses
Two little girls carry water in Zambia
Dear Friends,
Another year gone. Where did 2011 go? And at the same time, we thank God for all that was accomplished this year. Let us tell you how you have blessed many little children as you help them to secure their future through an education and other kinds of assistance:
• Our community school project for vulnerable and orphaned children called the WellSpring of Faith and Hope School or "WOFAH" (located in a poor urban area of the capitol city of Lusaka, Zambia) had an incredible year with your financial support and prayers. The children have had a stable educational year with classes every day on the school calendar, and we thank God that they have been (for the most part) happy and healthy, and have done well.
• A team of young people at Bloomsburg University donated school supplies to ensure the children have pencils, pens, erasers, and pencil cases just in time for the end of school year exams.
• With the help of our friends at Altrusa International in San Diego, and from Zamshu in Zambia, our WOFAH children received shoes just in time for the rainy season.
• The 2011 mission team from Second Baptist Church, Germantown, Pennsylvania blessed the children and us by coming and ministering through Vacation Bible School for several days as well as feeding the children daily healthy lunches.
• With help from our friends at the International School in Japan and the Istanbul International Community School in Turkey, we received seed-funds and started looking for land to build the WOFAH School so we can stop renting school rooms.
• Little girls in the weekly Bible Study Sarah leads received pillowcase dresses hand-sewn by some wonderful women in Laramie, Wyoming.
• Our friends at the American International School of Lusaka (AISL) have conducted professional and computer training with our community school teachers, as well as student exchanges between the AISL and WellSpring children in music, swimming and computer technology education.
This is just the beginning of the marvelous work that has begun in Zambia, and we really need your continued support. With your help and the help of our friends and partners in San Diego, Zambia, Japan, and Turkey a vital chain has been forged to help the children get an education, improve their lives, and change their community for the better. Your contributions and prayer support have made all this possible. Your contributions keep us on the field and allow us to continue to be the hands and feet of Christ.
For our current supporters, we need your ongoing support! We need new supporters to partner with us to continue these vital ministries. It is critical: we are at only 50 percent of our needed support level; and in order to remain in Zambia, we need more supporters so we can carry out this crucial work. Please consider an added or new "special end of year" or "beginning of the new year" donation that will make a difference and help to change lives.
We ask all who are part of our Missionary Partnership Network (MPN), and all those who wish to partner with us and become members of our MPN, to send your contributions to:
ABC/International Ministries
P.O. Box 851
Valley Forge, PA 19482-0851
-- with your check-note marked "For the support of the West's in Zambia". Also, you can contribute to our ministry on-line by entering an amount in the window at the top of this page and clicking on the "Give" button next to it.
At this wonderful and blessed time of the year, we thank you and pray God's richest blessings on you and yours.
In His Service,
Charles and Sarah West
MPT website: www.westinzambia.org


text:
Doctor Who Giant Hidden Tardis Mug
Out of stock
- Exquisitely moulded replica at the bottom of this Dr Who Mug
- The more you drink, the more the Tardis will appear
- This Dr Who memorabilia mug will look sit nicely on your side
- Fully licensed Dr Who product
- Ideal birthday gift, or treat for avid Dr Who fan
Back to Top
More product info
Do you find yourself wishing you were just like David Tenant from Doctor Who? Well, now you can travel time and explore the galaxies just like him! With this Giant Hidden Tardis Mug, you can simply drink up and watch a replica Tardis appears at the bottom of the mug!
Sit back and sip away as you day dream about defeating the Daleks and saving the day.
Please note: Not dishwasher safe


text:
The company began development on a line of Wii-based titles in the Summer of 2008, and had their first release, The Incredible Maze on Nintendo's WiiWare service October 24, 2008. Digital Leisure followed up this release in November 2008 with Sudoku Challenge!. On March 27, 2009, Digital Leisure released Texas Hold'em Tournament on the WiiWare service and plans to follow up these releases with other WiiWare and disc based titles in the near future.
Source: Wikipedia, "Digital Leisure", available under the CC-BY-SA License.