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Cyclic nucleotide–gated (CNG) channels are critical components in the visual and olfactory signal transduction pathways, and they primarily gate in response to changes in the cytoplasmic concentration of cyclic nucleotides. We previously found that the ability of the native rod CNG channel to be opened by cGMP was markedly inhibited by analogues of diacylglycerol (DAG) without a phosphorylation reaction (Gordon, S.E., J. Downing-Park, B. Tam, and A.L. Zimmerman. 1995. Biophys. J. 69:409–417). Here, we have studied cloned bovine rod and rat olfactory CNG channels expressed in Xenopus oocytes, and have determined that they are differentially inhibited by DAG. At saturating [cGMP], DAG inhibition of homomultimeric (α subunit only) rod channels was similar to that of the native rod CNG channel, but DAG was much less effective at inhibiting the homomultimeric olfactory channel, producing only partial inhibition even at high [DAG]. However, at low open probability (Po), both channels were more sensitive to DAG, suggesting that DAG is a closed state inhibitor. The Hill coefficients for DAG inhibition were often greater than one, suggesting that more than one DAG molecule is required for effective inhibition of a channel. In single-channel recordings, DAG decreased the Po but not the single-channel conductance. Results with chimeras of rod and olfactory channels suggest that the differences in DAG inhibition correlate more with differences in the transmembrane segments and their attached loops than with differences in the amino and carboxyl termini. Our results are consistent with a model in which multiple DAG molecules stabilize the closed state(s) of a CNG channel by binding directly to the channel and/or by altering bilayer–channel interactions. We speculate that if DAG interacts directly with the channel, it may insert into a putative hydrophobic crevice among the transmembrane domains of each subunit or at the hydrophobic interface between the channel and the bilayer.
Cyclic nucleotide–gated (CNG) ion channels play a vital role in visual transduction (for review see Roof and Makino 2000) and olfaction (for reviews see Dionne and Dubin 1994; Zufall et al. 1994; Menini 1995), and have been identified in numerous other tissues, including the heart (Biel et al. 1993; Ruiz et al. 1996) and kidney (Biel et al. 1994). CNG channels have been shown to consist of two to three (Bönigk et al. 1999) different types of subunits arranged in a tetrameric structure (for reviews see Kaupp 1995; Molday and Hsu 1995; Yau and Chen 1995; Zimmerman 1995; Finn et al. 1996; Zagotta and Siegelbaum 1996; Wei et al. 1998; Broillet and Firestein 1999). They are structurally homologous to voltage-gated ion channels (Jan and Jan 1992) with six transmembrane segments and a pore-forming loop between S5 and S6, but they are only weakly voltage-activated, and they open in response to cyclic nucleotides whose concentration varies with physiological status. Most CNG channels, including rod and olfactory types, are non-selective cation channels whose opening causes membrane depolarization and Ca2+ entry (for reviews see Kaupp 1995; Zimmerman 1995).
In rod cells, light activates an enzyme cascade that decreases the level of cytosolic cGMP, resulting in the closure of CNG channels that are normally open in the dark (for review see Roof and Makino 2000). In contrast, odorants activate another enzyme cascade that leads to the stimulation of adenylate cyclase and the production of cAMP, which opens olfactory CNG channels (for reviews see Dionne and Dubin 1994; Zufall et al. 1994; Menini 1995). Olfactory and rod CNG channels have similar properties, but olfactory channels are much more sensitive to cyclic nucleotides than are rod channels. Olfactory channels also respond to comparable concentrations of cAMP and cGMP, whereas rod channels are much more sensitive to cGMP than to cAMP (for reviews see Zimmerman 1995; Zagotta and Siegelbaum 1996). In fact, cAMP is only a partial agonist of the rod CNG channels. Furthermore, CNG channels are modulated by cellular factors, including phosphorylation enzymes (Gordon et al. 1992; Molokanova et al. 1997; Muller et al. 1998), Ca2+/calmodulin (Hsu and Molday 1993; Chen and Yau 1994; Gordon et al. 1995b), endogenous Ca2+ binding proteins (Gordon et al. 1995b; Balasubramanian et al. 1996), transition metal divalent cations (Idelfonse et al. 1992; Karpen et al. 1993; Gordon and Zagotta 1995a), and lipids, including diacylglycerol (DAG; Gordon et al. 1995a; Womack et al. 2000).
We previously reported that DAG modulation of the native CNG channel from frog rod outer segments is independent of phosphorylation (Gordon et al. 1995a). Evidence suggests that DAG also modulates other types of ion channels independently of protein kinase C, causing either activation or inhibition of channel function (Bowlby and Levitan 1995; Hofmann et al. 1999). In addition, DAG is a potential precursor of polyunsaturated fatty acids, such as arachidonic and linoleic acids, both of which have been shown to reversibly activate light-sensitive channels in Drosophila photoreceptors (Chyb et al. 1999). Thus, the production of DAG via activation of phospholipase C may have multiple ways of affecting channel behavior without relying on a phosphorylation pathway. Interestingly, a recent study of mammalian rod CNG channels (α and β subunits) expressed in Xenopus oocytes has shown that long-chain DAG is stimulatory, whereas the cellular precursor to DAG, PIP2, is inhibitory. However, PIP2 inhibition is not as strong when only α subunits are expressed (Womack et al. 2000). Although a physiological role for DAG in the visual or olfactory pathway remains undetermined, in the present study, DAG is used as a tool to dissect the functional differences of the rod and olfactory CNG ion channels.
To elucidate the mechanism of DAG inhibition of CNG channels, we explored the effect of a short-chain DAG analogue on cloned rod and olfactory channels expressed in Xenopus oocytes. Rod channels exhibited greater inhibition than olfactory channels at saturating cGMP concentrations. However, DAG inhibition was much more efficient at low open probabilities for both channel types, suggesting that DAG stabilizes the closed states of the channel. In addition, the Hill coefficients from DAG dose–response curves suggested that multiple DAG molecules participate in the inhibition of a channel.
Because these two CNG channels showed differences in their inhibition by DAG, we also examined the DAG modulation of a series of chimeras of the rod and olfactory channels (Gordon and Zagotta 1995b; Fodor et al. 1997). Our findings suggest that the sequence differences in the transmembrane segments and loop structures, rather than those in the amino and carboxyl termini, may be responsible for the differences in DAG inhibition between the two channels. At saturating cGMP, Hill coefficients for DAG inhibition ranged from 1.5 to 2.8 for this series of chimeras, indicating that its mechanism of inhibition must differ from that of tetracaine, which demonstrates a Hill coefficient of ∼1 and shows obvious voltage dependence (Fodor et al. 1997). We propose that DAG stabilizes the closed states of the channels either by direct interaction with the channel protein, by distortion of bilayer–channel interactions, or by some combination of these mechanisms.
Materials And Methods
Expression of Channels in Xenopus Oocytes
The plasmids containing the α subunits of bovine rod (CNG1), rat olfactory (CNG2), and chimeric cDNA were provided by William N. Zagotta (University of Washington, Seattle, WA). See Richards and Gordon 2000 for other terminology for these channels. The olfactory α subunit clone originated from the laboratories of R.R. Reed (Johns Hopkins University, Baltimore, MD) and R.S. Molday (University of British Columbia, Vancouver, British Columbia, Canada). Construction of the chimeras was previously described in Gordon and Zagotta 1995b,. The plasmid containing the mouse olfactory clone was provided by Maria Ruiz (Entelos, Inc., Menlo Park, CA) (Ruiz et al. 1996), and this channel has been shown to behave similarly to the rat olfactory channel (Crary et al. 2000); in this study, the wild-type mouse olfactory channel was only used for the single-channel analysis. The vectors all contain the untranslated sequence of the Xenopus β-globin gene, which promotes high protein expression in oocytes (Liman et al. 1992). Capped cRNA was made by in vitro transcription using either Promega's RiboMAX™ large-scale RNA production system or Ambion's mMessage Machine.
Sections of ovaries containing oocytes were surgically removed from anesthetized Xenopus laevis frogs, and individual oocytes were isolated from clumps of tissue by treatment with 1 mg/ml collagenase type 1A (Sigma-Aldrich) in a buffer containing low calcium (82.5 mM NaCl, 2.5 mM KCl, 5 mM HEPES, and 1 mM MgCl2, pH 7.6). The cRNA was injected into Xenopus oocytes using a Drummond Nanoject oocyte injector. The injected oocytes were initially incubated at 15°C overnight, and then at 18°C for 1–10 d before testing for expression via the patch-clamp technique. For single channel studies, the oocytes were stored at 4°C once a desirable level of expression was reached. The saline solution for maintaining oocytes contained the following: 96 mM NaCl, 2 mM KCl, 1.8 mM CaCl2, 5 mM HEPES, 1 mM MgCl2, 2.5 mM pyruvic acid, 100 U/ml penicillin, and 100 μg/ml streptomycin, pH 7.6. The vitelline membrane was removed by treatment with a hypertonic solution (200 mM N-methyl-d-glucamine, 2 mM KCl, 10 mM EGTA, 10 mM HEPES, and 1 mM MgCl2, pH 7.4) and mechanical dissection to expose the plasma membrane for patch-clamp recording.
Application of Electrophysiological Solutions
The oocyte chamber for patch-clamp experiments was a plastic petri dish, and water-soluble solutions were applied using a 36-solution patch perfusion system, the RSC-100 rapid solution changer (Molecular Kinetics). Solutions consisted of the following: 130 mM NaCl, 0.2 mM EDTA, and 2 mM HEPES buffer, pH 7.2, with varying concentrations of cGMP (Sigma-Aldrich) and cAMP (Calbiochem-Novabiochem). When necessary, 500 μM niflumic acid (Sigma-Aldrich) was added to the extracellular solution to block Ca2+-activated Cl− channels, which are endogenous to the oocyte membranes. The DAG analogue, 1,2-dioctanoyl-sn-glycerol (1,2-DiC8; Sigma-Aldrich) was dissolved in DMSO (Sigma-Aldrich) at 0.5–20-mM stock concentrations and stored at −20°C. Because of the lipophilicity of the analogues, the DAG solutions were not applied to the patch through Teflon tubing because we found that this resulted in a decrease in the response, as though the DAG had adhered to the tubing, effectively reducing the concentration of the DAG before its exposure to the patch. Instead, the DAG analogue was applied to the intracellular surfaces of patches by direct addition to the bathing solution in the petri dish, where it was carefully and thoroughly mixed using a transfer pipet. The petri dish and agar bridge were replaced after each experiment with DAG.
Electrophysiological Recordings and Analysis
Data were collected from inside-out patches in the steady state after any spontaneous changes in cGMP affinity, which usually resulted in a left shift of the cGMP dose–response curve. These changes have been attributed to dephosphorylation by endogenous patch-affiliated phosphatases (Gordon et al. 1992; Molokanova et al. 1997), and were monitored here by sampling the current periodically at a low level of cGMP (below the K1/2) while incubating the patch in saturating cGMP and waiting for the monitored current to stabilize. The time required for this stabilization ranged from ∼10 to 40 min, and the magnitude and time course of the effect varied with the channel type.
For DAG dose–response curves, the bath contained a saturating concentration of cGMP (2 mM for rod and chimeric channels and 100 μM for olfactory), and DAG was added in increments resulting in changes in bath concentration ranging from 0.05 to 48 μM. However, the DMSO concentration remained below 1.2%, which had no effect on the channels or the leak current. The current was monitored for several minutes after each addition of DAG to insure that the effect of the DAG had stabilized; stabilization was usually complete within <1 or 2 min. This time may have been required for DAG to insert into the membrane, and the reversal of the DAG effect was also slow as reported previously (Gordon et al. 1995a). Although we cannot measure the final concentration of DAG in the membrane after this stabilization, we were working well below the critical micelle concentration (270 μM) of a similar hydrophobic molecule (1,2-dioctanoyl phosphatidylcholine; Marsh and Phil 1990) and below the IC50 reported for DAG inhibition of peptide-induced polarity of neutrophils (Zimmermann et al. 1988), indicating that we are in the linear concentration range during our DAG dose–response experiments. In fact, the DAG dose–response curve for CHM3 provides evidence that the concentration of DAG is increasing incrementally up to at least 6 μM (see Fig. 7), confirming the availability of soluble DAG for the observed partial response of the olfactory channel in this same range.
For experiments with the native channel, rod cells were obtained from Rana Pipiens according to an established method (Zimmerman and Baylor 1992). Excised inside-out patches from rod outer segments were studied using the same methods and solutions as for the Xenopus oocytes. Pipet openings were 0.5–20 μm in diameter and had resistances of 0.6–15 MΩ. All recordings were obtained at room temperature (20–25°C). Recordings of the current were made as the voltage was changed from −100 to +100 mV in steps of 20 or 50 mV, from a holding potential of 0 mV. The leak currents obtained in the absence of cGMP were subtracted from each record. For single-channel studies, three to five continuous 1-s recordings were made at +80 mV.
Sampling (digitization) of currents was accomplished using an Axopatch 1B, 1C, or 200 patch-clamp amplifier (Axon Instruments) with analogue-to-digital converters, which sent output to a Macintosh Quadra computer running Pulse software (Instrutech). Before digitization, the data were low pass–filtered using a selectable 8-pole Bessel filter (Frequency Devices). The filter cutoff frequency (−3 dB point) was 2 kHz for multichannel patches and 5 kHz for single-channel patches. Sampling rates were set at five times the filter cutoff frequency to prevent aliasing. Data analysis was performed with the graphical analysis software IgorPro (WaveMetrics).
DAG Inhibition of Rod Channels Is Greater than that of Olfactory Channels
Fig. 1 demonstrates that DAG has a greater inhibitory effect on rod channels than on olfactory channels and that both channel types are more sensitive to DAG at low concentrations of agonist. Electrophysiological recordings were obtained from inside-out excised patches from Xenopus oocytes expressing α homomultimers of bovine rod or rat olfactory CNG ion channels. Fig. 1 shows the current traces recorded for each wild-type channel at low and high cGMP concentrations in both the presence and absence of 0.5 μM DAG. At saturating cGMP concentrations, this low dose of DAG had a small effect on the olfactory channel, but decreased the current of the rod channel by ∼42%. At lower cGMP concentrations (near the K1/2 for each channel), the same amount of DAG reduced the current for both rod and olfactory channels to a greater extent. This suppression was found to be reversible (data not shown), which is consistent with the reversible suppression observed for the native rod channels (Gordon et al. 1995a). Furthermore, this inhibition by DAG occurred in the absence of ATP, demonstrating that DAG cannot be acting through phosphorylation by protein kinase C. In addition, consistent with the original study (Gordon et al. 1995a), 1,3-DiC8 was just as effective as 1,2-DiC8, and 1,2-dioctanoyl ethylene glycol was much less effective (data not shown).
Fig. 2 illustrates a more quantitative comparison of the response of the two channels to DAG. Dose–response curves for activation of each channel by cGMP demonstrate a large shift to the right in the presence of 0.5 μM DAG. For the rod channel, the K1/2 increased by a factor of 3.2, from 24 to 76 μM cGMP, and for the olfactory channel, the K1/2 increased by a factor of 2.2, from 1.8 to 4.0 μM cGMP. For the rod channel, there is also a significant (29%) reduction in the maximal current with DAG, whereas this reduction is much smaller (3%) for the olfactory channel. The Hill coefficients remained essentially unchanged for both channels.
Previous results showed that application of excess cyclic nucleotide did not reverse the inhibition of the native rod channel by DAG, suggesting that the lower apparent affinity does not result from competitive inhibition of agonist binding (Gordon et al. 1995a). Similar findings in the current study suggest that this is also the case for the cloned channels. This is apparent for the rod channel in Fig. 2 A: increasing [cGMP] beyond saturation does not induce recovery of the DAG-suppressed current. A similar result was observed with the olfactory channel when a higher DAG concentration was used to produce a greater decrease in maximal current at high cGMP concentrations (data not shown).
The dose–response curves for DAG at saturating [cGMP] are dramatically different for the rod and olfactory channels. Fig. 3 indicates that rod cGMP-activated currents are completely suppressed by ∼3 μM DAG at saturating (2 mM) cGMP, but the olfactory cGMP-activated currents show only partial suppression at saturating (100 μM) cGMP, even with DAG concentrations as high as 12 μM. In fact, the fractional suppression of the olfactory channel remained at this low level even when we raised the DAG concentration to 48 μM (data not shown). For the rod channel, the IC50 for inhibition was 0.83 μM DAG, and the Hill coefficient was 1.7. However, for the olfactory channel, the concentration required to obtain half maximal inhibition was 3.24 μM DAG, with a Hill coefficient of 1.4. Thus, there is approximately a fourfold difference between the DAG IC50 obtained for the rod channel and that obtained for the olfactory channel at saturating [cGMP].
Fig. 3 also demonstrates the higher sensitivity of both channels to DAG at lower concentrations of cGMP. The DAG dose–response curve from a representative patch at low [cGMP] is shown for each channel. In comparison to the corresponding DAG dose–response curve at high [cGMP], the curve at low [cGMP] is shifted to the left for both channels, and, interestingly, for the olfactory channel, the inhibition reaches 100%. The I/Imax for both channels was ∼0.40 before the addition of DAG; the IC50 values for inhibition were 0.41 and 0.47 μM, and the Hill coefficients were 1.3 and 1.4 for the rod and olfactory channels, respectively. Furthermore, for the rod channel, DAG inhibition at saturating [cAMP] was comparable to that at a low cGMP concentration that produced a similar fractional activation (data not shown). This finding suggests that DAG preferentially affects closed channels, regardless of ligand occupancy.
For the native rod channel, the DAG IC50 was previously reported to be 22 μM, which is quite different from the value reported here for the cloned rod homomultimer. Although preliminary studies with the cloned channel resulted in development of a new method of administering DAG to the patch without the use of Teflon (see materials and methods), we could not ignore the possibility that the cloned channel might be more sensitive to the DAG analogue than is the native channel. Thus, we reexamined the native channel using our new DAG application technique. Our data confirmed that the response of the native channel to the DAG analogue is similar to that for the cloned channel, as shown by the triangles in Fig. 3. This indicates that the values from the previous study are in error, most likely because of the adherence of DAG to the Teflon delivery tubes, effectively reducing the concentration of DAG in solution.
Reductions in Maximal Current Are Due to Decreasing Open Probability
Single-channel analysis was performed to determine whether the reduction in the maximal current by DAG was due to a decrease in open probability (Po) or in single-channel conductance. Fig. 4 and Fig. 5 show single-channel currents and amplitude histograms at +80 mV for rod and olfactory channels in the presence and absence of DAG. The patch in Fig. 4 contained two rod channels (see legend for note about sequence); the patch in Fig. 5 contained a single wild-type olfactory channel. In the presence of saturating cGMP, both rod and olfactory channels spent most of their time in the open state. With the addition of DAG to the bath, both types of channels spent more time closed. Furthermore, the single-channel amplitude remained unchanged by the DAG, at 2.1 pA for the rod channel and 3.2 pA for the olfactory channel. Thus, DAG causes a reduction in Po, but does not appear to change the single-channel conductance.
DAG Inhibition Is Independent of Voltage
The addition of DAG at saturating cGMP alters the steady state current–voltage relations of both channels in a manner that reflects the lower Po of each channel in the presence of DAG, suggesting that DAG does not impart its own voltage dependence but merely conveys the gating behavior that is apparent when fewer channels are open. In Fig. 6, the current–voltage relations for each channel are shown under three different experimental conditions: at saturating cGMP, both in the presence and absence of 2 μM DAG, as well as at a low cGMP concentration. For the rod channel, the addition of DAG at saturating cGMP creates a current–voltage relation similar to that observed when the channel responds to 10 μM cGMP, which is below the K1/2 value for this patch. These relations show the characteristic increase in rectification seen for the native rod channel at low cGMP concentrations (Karpen et al. 1988). The current–voltage relation of the olfactory channel in the presence of 2 μM DAG at saturating cGMP correlates with the current–voltage relation observed when the channel responds to 5 μM cGMP, which is above the K1/2 for this channel, reflecting the smaller effect of DAG on the olfactory channel. These current–voltage relations indicate that the only voltage dependence of DAG inhibition is that intrinsic to normal gating. This is in contrast to the voltage-dependent block observed with tetracaine (Fodor et al. 1997).
Structural Localization of Channel Region(s) Influencing Differential DAG Inhibition of Rod and Olfactory Channels
We used chimeras of rod and olfactory channels in an attempt to identify channel regions responsible for the differential inhibition of these two channels by DAG. For each chimera, we measured the K1/2 for activation by cGMP, the maximal percent inhibition by DAG at saturating cGMP and the DAG IC50 for this level of inhibition. Data were averaged from three to six patches for each chimera. Fig. 7 shows DAG dose–response curves measured at saturating cGMP for three representative chimeras, CHM3, CHM11, and CHM13 (see Fig. 8 for structures). Interestingly, although CHM3 contains the amino and carboxyl termini of the rod channel, its response to DAG is more like that of the olfactory channel; it is only partially inhibited by DAG and its IC50 (3.30 μM) is closer to that of the olfactory channel than to that of the rod channel. These data provided the first hint that the termini are not as important as the transmembrane segments and loops in determining the differential effects of DAG on rod and olfactory channels. Additional support for this notion comes from experiments with a mutant of the rod channel, H468Q. This mutant channel has dramatically increased cAMP efficacy (with saturating cAMP producing ∼40% of the maximal cGMP-activated current, as compared with only one to a few percent for wild-type rod channels). However, its DAG IC50 (0.33 μM, mean from 11 patches; data not shown) is not very different from that of the wild-type rod channel, and it is fully inhibited by DAG at saturating cGMP. Thus, functionally significant mutations in the carboxyl terminus have rather insignificant effects on inhibition by DAG.
Fig. 8 shows the values of cGMP K1/2 and of DAG IC50 at saturating cGMP for each chimera and the two wild-type channels. The results are displayed, along with the chimera structures, in the order of increasing cGMP K1/2, which reflects the relative ease of opening of each chimera. Similar to the data for tetracaine inhibition observed by Fodor et al. 1997, the DAG IC50 and cGMP K1/2 are roughly inversely correlated. However, this inverse correlation does not always strictly hold, as demonstrated in the companion paper (Crary et al. 2000, in this issue).
The corresponding maximum inhibition by DAG at saturating cGMP for each chimera and the wild-type channels is shown in Fig. 9, presented in the order of increasing maximum inhibition. The chimeras that reveal only partial inhibition tend to contain more olfactory-like sequences in the core regions of the channel, from S2 through S6 including the interconnecting loops of the transmembrane segments. In general, differences in the amino and carboxyl termini do not explain the differential DAG inhibition of the rod and olfactory channels; however, it appears that the presence of both amino and carboxyl termini from the rod in CHM3 may influence this chimera's behavior, increasing the maximum inhibition toward that of the rod channel.
Although extensive research has been performed on CNG channels, the molecular mechanism of gating is not fully understood. By exploring the molecular mechanism of DAG inhibition of these channels, we may further understand the complex gating of these ion channels and, in particular, the differences between the properties of the rod and olfactory channels. In this study, we have compared DAG inhibition of the bovine rod and rat olfactory α homomultimeric channels expressed in Xenopus oocytes. For both channels, inhibition by DAG is greater at low than at high channel Po, indicating that DAG stabilizes the closed states of the channels (Fig. 1 and Fig. 3). This finding is consistent with that previously reported for DAG inhibition of the native CNG channel in amphibian rod cells (Gordon et al. 1995a). Single channel data (Fig. 4 and Fig. 5) support the notion that DAG inhibits the channels by lowering open probability. As before, the experiments were performed without the addition of ATP, indicating that the mechanism does not involve channel phosphorylation as a result of DAG activation of PKC. DAG inhibition of the cloned channels is also voltage-independent, with changes in current–voltage relations merely reflecting the lower Po in the presence of DAG.
We find that rod CNG channels are much more strongly inhibited by DAG than are olfactory channels. This is especially apparent at saturating cGMP concentrations, where the rod cGMP-activated currents are completely abolished by DAG, but the olfactory currents are only partially suppressed even at high concentrations of DAG (Fig. 3). This finding suggests that the olfactory channel, whose opening is more energetically favored (Gordon and Zagotta 1995b), is less sensitive to inhibition by DAG. This is reminiscent of the phenomenon observed for inhibition of these two channels by tetracaine (Fodor et al. 1997).
Our previous work with the native rod CNG channel suggested that DAG may be a closed state inhibitor (Gordon et al. 1995a). Here, we have more thoroughly addressed this question by comparing DAG inhibition of the rod and olfactory channels. In Fig. 1 and Fig. 2, the larger reduction in current at lower cGMP concentrations suggests that DAG stabilizes the closed or unliganded states of each channel. Furthermore, DAG has a much larger effect at saturating concentrations of the partial agonist (cAMP; data not shown) than at saturating cGMP, indicating that it is the Po, rather than ligand occupancy, that matters. Fig. 3 provides further evidence that DAG is a closed state inhibitor. At low [cGMP], the DAG IC50 is decreased for both channels in comparison to the IC50 at saturating cGMP. In addition, the olfactory channel can be fully inhibited at low [cGMP] when the channels are more frequently in the closed state. Finally, Fig. 2 indicates that DAG is not a competitive inhibitor of cGMP binding, since large doses of cGMP cannot reverse the effects of DAG. These data are consistent with a mechanism in which DAG disturbs the allosteric opening transition of the channel.
To explore further the behavior of the channels in response to DAG, we also examined patches containing only one or two channels. Single-channel analysis revealed a DAG-induced decrease in Po without a measurable change in single-channel conductance (Fig. 4 and Fig. 5). Hence, the reduction in the maximum current at saturating cGMP most likely was not due to the incomplete block of the pore by DAG, resulting in channel openings of lower current amplitude. The Hill coefficients from the DAG dose–response curves suggest that if DAG were to act by blocking the pore, it must do so with more than 1 molecule per channel. It appears that at least two DAG molecules are necessary for efficient inhibition of each channel (Fig. 3), raising the question of whether one DAG molecule interacts with each of the four channel subunits.
Since cloned rod and olfactory channels are differentially inhibited by DAG, we used a series of chimeras of these two channels in an attempt to attribute these differences in inhibition to specific regions of the channels. Chimeras have offered a valuable approach to understanding the differences in rod and olfactory channel behavior with respect to apparent agonist affinity, efficacy, and tetracaine inhibition (Goulding et al. 1994; Gordon and Zagotta 1995b; Fodor et al. 1997). In this study, the series of chimeras from the Zagotta lab was examined with respect to cGMP K1/2 (without DAG), DAG IC50 and maximal percent inhibition observed at saturating cGMP concentrations. From the chimera data, the core region of the channel seems to play a greater role in determining the differential DAG inhibition of the rod and olfactory channels than do the amino and carboxyl termini. Our chimera results demonstrate an approximately inverse relationship between cGMP K1/2 and inhibitor IC50, like that seen for inhibition by tetracaine (Fodor et al. 1997). However, we have recently found that a point mutation in the S2–S3 olfactory loop produces dramatic changes in DAG and tetracaine sensitivity with only minimal changes in apparent agonist affinity (Crary et al. 2000, in this issue).
Since PKC activation is not involved, DAG may have a direct inhibitory interaction with the CNG channels themselves or act on an unknown channel modulator that does not require the addition of nucleoside triphosphates. A previously defined DAG binding motif in protein kinase C and RasGRP contains repeats of histidine and cysteine residues (Ebinu et al. 1998). We could not identify such a consensus site in the primary sequence of the rod or olfactory CNG channel. However, the required residues for a consensus site may be present in distant parts of the primary sequence, and the repeat sequences may evolve upon folding. The DAG binding site could also be formed through coordination of residues from neighboring subunits of the tetramer, as proposed for the formation of a Ni2+ binding site through coordination of H420 from adjacent subunits in these channels (Gordon and Zagotta 1995a,Gordon and Zagotta 1995c). Alternatively, it is possible that CNG channels may have a binding site that differs from the documented consensus site.
Our chimera study suggests that the transmembrane segments and attached loops may be important in determining the differential effects of DAG on rod and olfactory channels. Given the hydrophobic nature of DAG, the transmembrane segments are a likely target for this inhibitor. Furthermore, the spatial arrangement of the transmembrane segments may be critical for proper channel function. In other channels, rotation or twisting of these helical segments has been proposed for the allosteric transition from the closed to open state (Unwin 1995; Perozo et al. 1999). Hence, DAG may act as a wedge that inserts into an intracellular crevice between adjacent transmembrane segments, thereby preventing pore expansion via movements of the core helices. Alternatively, once in place, the head group may interfere with the normal association of the intracellular loops, the amino terminus, or the carboxyl terminus with the membrane surface, which might occur during gating. The structure of the connecting loops may also be critical, providing flexibility or rigidity to attached transmembrane domains and possibly determining the overall structure of the channel. Future studies using chimeras and mutations involving these regions may shed light on how DAG inhibits the conformational changes associated with the gating process and whether DAG, indeed, interacts directly with the channels.
In Fig. 10, we present two working models that represent possible mechanisms of DAG inhibition of olfactory and rod channels. Both models depict the two channels in either open or closed configurations (with and without DAG) at saturating agonist concentrations. The horizontal transitions in each model represent allosteric state changes, whereas vertical transitions represent binding or removal of multiple molecules of DAG. The top part of each diagram portrays the preference of both channel types for the open state in the absence of DAG, and the tendency for rod channels to be closed more of the time than are olfactory channels. For the rod channel, DAG strongly stabilizes the closed state, even under saturating agonist conditions. However, for the olfactory channel, the transition to the open state is still highly favored, even in the presence of DAG. This representation is based on the data showing that even high concentrations of DAG can only partially inhibit the olfactory channel at saturating [cGMP] (Fig. 3).
In the first model (Fig. 10 A), DAG is shown bound to a putative crevice on the intracellular side within the hydrophobic core of the channel. Alternatively, DAG may directly bind the channel at the hydrophobic interface between the transmembrane segments and the bilayer. An equally plausible mechanism (Fig. 10 B) for the action of DAG is that it inserts into the lipid bilayer, perturbing the coupling between the hydrophobic core of the bilayer and the hydrophobic membrane-spanning regions of the channel. This kind of alteration has been shown to modulate other channels (Lundbaek et al. 1996, Lundbaek et al. 1997; Lundbaek and Andersen 1999). For example, lysophospholipids have been shown to dramatically increase the dimerization constant for gramicidin (Lundbaek and Andersen 1994). Interestingly, bilayer deformation effects can occur without changes in membrane fluidity, and can even show apparent cooperativity, with large Hill coefficients (Cantor 1999), as observed with specific binding reactions (and as reported in this study).
The cartoon in Fig. 10 B shows one possible scenario by which DAG may alter bilayer–channel interactions. Here, the closed states are drawn distinctly shorter than the open states to suggest a smaller hydrophobic exterior surface of the protein. The surrounding bilayer is drawn with more curvature in the presence of DAG, to suggest a thinner, better matched, hydrophobic core at the interface between the bilayer and the closed channel. Thus, the addition of DAG is predicted to change the energetics of the channel conformational changes, thereby altering the open and closed probabilities. Of course, DAG may regulate the channels by some combination of the mechanisms depicted in Fig. 10 (A and B). The actual physical changes that occur in channel gating remain to be determined, as does the detailed mechanism by which DAG alters the conformational equilibrium. However, it would seem that hydrophobic inhibitors such as DAG may prove useful in reporting intramolecular interactions and movements in the hydrophobic core of channels.
We thank Drs. Sharona Gordon and Gary Yellen for helpful discussions, and Dr. William N. Zagotta for comments on an earlier version of the manuscript. We thank Dr. Maria Ruiz for the mouse olfactory clone and Dr. William N. Zagotta for the bovine rod and rat olfactory clones and the series of chimeric constructs. We are also grateful to Roberto Neisa and Melody Chang for technical assistance, and Joe Zimmerman for software development.
This work was supported by grants from the American Heart Association of Rhode Island (No. 97-07721S) and the National Institutes of Health (No. EY07774).
Abbreviations used in this paper: CNG, cyclic nucleotide–gated; DAG, diacylglycerol; Po, open probability.

domain: literature


Ninety-eight trainees from the Institute of Microbiology of the Chinese Academy of Sciences, the Chinese Academy of Inspection and Quarantine, the Chinese Center for Disease Control and Prevention (China CDC), and other relevant units participated in the opening ceremony of the 2022 high-level pathogenic microorganism laboratory biosafety training course, organized by the China CDC, on Aug 30.
Liu Dengfeng, commissioner of the Department of Health Science, Technology and Education of the National Health Commission, emphasized the requirements for laboratory biosafety in the Biosafety Law and said that high-level pathogenic microorganism laboratories play an important role in the identification of infectious disease pathogens, the establishment of animal models, and the research into detection reagents, medicines and vaccines. Liu also urged intensified management on high-level laboratory construction and biosafety.
Shi Jingjing, deputy director of the Science and Education Department of the Beijing Municipal Health Commission, said that the Beijing Municipal Health Commission will fully support the China CDC in construction of the high-level laboratory biosafety training base.
Guo Chunmei, deputy director of the Changping District Health Commission in Beijing, said that the commission also hopes that the China CDC will give full play to its advantages in the construction and management of pathogenic microorganism laboratories and promote the development of other laboratories in the district.
Liang Bing, director of the Biosafety Division of the Department of Health Science, Technology and Education of the National Health Commission, and Wu Guizhen, chief biosafety expert of the China CDC, jointly inaugurated the training base.


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|Bid||0.00 x 1300|
|Ask||0.00 x 800|
|Day's Range||112.18 - 114.10|
|52 Week Range||64.15 - 141.60|
|Beta (3Y Monthly)||1.27|
|PE Ratio (TTM)||32.48|
|Forward Dividend & Yield||1.48 (1.45%)|
|1y Target Est||N/A|
MainStay Large Cap Growth Fund is one of the best mutual funds. Its managers know where they like to look for growth stocks — and where not to bother.
Mad Money viewers Tuesday night were cautioned by Jim Cramer not to expect a trade deal with China at the G20 summit in Japan next week. Cramer said Broadcom Inc. In the daily bar chart of AVGO, below, we have seen a sharp May decline which included a gap to the downside.
Despite the S&P 500 trading at lofty heights, 8 blue chip stocks are still selling well below their record highs but may be poised for big gains as investors pursue diverse strategies to profit amid a volatile U.S.-China trade war. Some professional investors are wading in to buy beaten down blue chips from the semiconductor industry, including Broadcom Inc. (AVGO), Xilinx Inc. (XLNX), NXP Semiconductors N.V. (NXPI) and Skyworks Solutions Inc. (SWKS). Goldman says these service stocks are much more likely to outperform than stocks in goods-producing sectors.
The Trump administration has ordered American companies not to do business with Huawei, claiming the Chinese company represents a threat to national security.
Major U.S. chip makers, including Intel Corp. , Qualcomm Inc. and Xilinx Inc. , have quietly lobbied the Trump administration to ease its ban on sales to Chinese tech giant Huawei Technologies Co., Reuters reported Sunday. One source told Reuters that the aim was not to help Huawei, but to prevent harm to U.S. companies. Reuters said Huawei spent about $11 billion buying components from U.S. companies in 2018. The Trump administration announced the ban in May, after trade negotiations between the U.S. and China stalled. Earlier this month, the acting White House budget chief sought to delay implementation of the ban, the Wall Street Journal reported, arguing that the burden would fall on U.S. companies that did business with Huawei.
SAN FRANCISCO/WASHINGTON, June 16 (Reuters) - Huawei's American chip suppliers, including Qualcomm and Intel, are quietly pressing the U.S. government to ease its ban on sales to the Chinese tech giant, even as Huawei itself avoids typical government lobbying, people familiar with the situation said. Executives from top U.S. chipmakers Intel and Xilinx Inc attended a meeting in late May with the Commerce Department to discuss a response to Huawei's placement on the black list, one person said. The ban bars U.S. suppliers from selling to Huawei, the world's largest telecommunications equipment company, without special approval, because of what the government said were national security issues.
Broadcom (AVGO) presented their second quarter earnings on Thursday after market close, and the call, as described by CNBC's Jim Cramer, was "truly depressing."
Investing.com - The S&P; 500 closed flat on Friday as firmer retail sales pointing to underlying strength in the U.S. economy were countered by a slump in chip stocks, led by Broadcom.
Demand for next-generation 5G devices is on the rise, according to DigiTimes: Taiwan-based integrated circuit design houses are seeing a slowing pace in 4G orders, the publication said Friday. U.S. stocks ...
Investing.com - Broadcom fell sharply Friday, sending chip stocks lower after the company delivered an ominous outlook and reported revenue that fell short of estimates.
Several semiconductor companies saw shares decline in after-hours trading Thursday following Broadcom Inc.'s earnings report, which suggested a second-half rebound for chips looks less likely. Among stocks falling more than 1% immediately after Broadcom reported were Texas Instruments Inc. , Xilinx Inc. , Skyworks Solutions Inc., Qualcomm Inc. , Qorvo Inc. and Nvidia Corp. Larger chip makers like Intel Corp. , Advanced Micro Devices Inc. and Micron Technology Inc. fell by smaller amounts, closer to 0.5%, though action was jagged amid high volume for the extended session. Broadcom cut its fiscal-year forecast in its second-quarter report after trading closed Thursday, and Chief Executive Hock Tan said in a statement that he sees "a broad-based slowdown in the demand environment." "As a result, our customers are actively reducing their inventory levels, and we are taking a conservative stance for the rest of the year," Tan said. Chip companies earlier this year said that a slowdown would reverse by the second half of the year, but returns since then have created doubts about the timeline. Broadcom shares were down about 7% in after-hours trading.
Xilinx, Inc.'s (NASDAQ:XLNX) latest earnings update in March 2019 signalled that the company gained from a sizeable...
“I don’t believe what’s been weighing on Chinese equities has been the trade war,” said Noah Blackstein, who manages C$5.1 billion ($3.8 billion) at Dynamic Funds, a unit of Bank of Nova Scotia. Crackdowns on new video games and social media are just two examples of unexpected regulatory changes that had a “material” impact on the profit of Chinese companies last year, said Blackstein, 49, who reduced his exposure to stocks like Alibaba Group Holding Ltd. and Tencent Holdings Ltd. as a result. Blackstein manages the C$1.4 billion Dynamic Power Global Growth Class fund which has outperformed 1,083 global peers that have assets of more than $100 million, returning 388% over the past decade, according to data compiled by Bloomberg.
Popular technology stocks have recently staged a rebound. But segmented money flows show the rally is suspect. Let’s examine the issue with the help of a chart. Chart Please click here for a chart showing money flows in 11 popular tech stocks.
Chipmaker Nvidia is at the forefront of AI and machine learning, but earnings and share prices have dived. Here is what fundamental and technical analysis say about buying Nvidia stock now.
Xilinx Inc NASDAQ/NGS:XLNXView full report here! Summary * Bearish sentiment is low Bearish sentimentShort interest | PositiveShort interest is low for XLNX with fewer than 5% of shares on loan. The last change in the short interest score occurred more than 1 month ago and implies that there has been little change in sentiment among investors who seek to profit from falling equity prices. Money flowETF/Index ownership | NeutralETF activity is neutral. ETFs that hold XLNX had net inflows of $2.48 billion over the last one-month. While these are not among the highest inflows of the last year, the rate of inflow is increasing. Economic sentimentPMI by IHS Markit | NeutralAccording to the latest IHS Markit Purchasing Managers' Index (PMI) data, output in the Technology sector is rising. The rate of growth is weak relative to the trend shown over the past year, however. Credit worthinessCredit default swapCDS data is not available for this security.Please send all inquiries related to the report to firstname.lastname@example.org.Charts and report PDFs will only be available for 30 days after publishing.This document has been produced for information purposes only and is not to be relied upon or as construed as investment advice. To the fullest extent permitted by law, IHS Markit disclaims any responsibility or liability, whether in contract, tort (including, without limitation, negligence), equity or otherwise, for any loss or damage arising from any reliance on or the use of this material in any way. Please view the full legal disclaimer and methodology information on pages 2-3 of the full report.
Not too long ago, Nvidia (NASDAQ:NVDA) stock could do no wrong. At the highs, Wall Street experts unanimously said that it was the stock to own in every portfolio. Nvidia hit $290 per share at its all-time high.Source: Shutterstock Since then, Nvidia stock has fallen off a cliff. It's now under $150 and can't even hold a rally. In March it showed some promise, but that too failed. The bulls couldn't even fill a giant open gap from its horrendous November earnings report.I recently wrote a note that it could be headed to $200 per share, but since then, that effort also failed. Now it's back to the December lows while the S&P 500 remains 21% off its lows. Clearly NVDA stock is out of favor on Wall Street.InvestorPlace - Stock Market News, Stock Advice & Trading Tips Looking Forward in NVDA StockSo what now? Is it time to panic out of it?No.If I still am long Nvidia at this level, it would be a mistake to capitulate right here. This is a proven bounce level for the stock, so I can probably count on it continuing to be so until it's lost. Technically speaking, the stock has been in a descending lower high trend bouncing against a proven floor. More often than not, the floor holds, making a rally from these level likely. To open a new trap door from these low levels, I bet it will take incrementally bad news. * 7 S&P 500 Dividend Stocks to Buy at Least Yielding 3% But the risk is there, so it's important to know where it lies. If the bears are able to breach this support, then they would trigger a pattern to target $98 per share. This is not my forecast, but it is the negative scenario that looms below.NVDA valuation helps the bulls here. At $290 per share it was too expensive, as the price crash proves. But at these levels here the trailing price-to-earnings ratio is down to 30. This is relatively cheap if I compare it to something like Advanced Micro Devices (NASDAQ:AMD). It is still almost three times as expensive as Intel (NASDAQ:INTC) -- but for good reason. But maybe you get what you pay for, as AMD is up almost 80% year to date, while for the same period NVDA is up 8% and INTC is red.Fundamentally speaking, the demand on Nvidia products and services will continue to be strong for years to come. This is a premier technology company and one of only a few that will power the tremendous global migration to the digital world. We have a new world of AI and self-driving cars coming soon. This trend is not likely to reverse anytime soon.Nvidia's technology is solid, so they show no evidence of managerial flubs. So as an investor, I continue to give them the benefit of the doubt that they will continue to successfully execute on their plans.So to recap so far, here are my assumptions: I acknowledge that NVDA is no longer popular on Wall Street, but that alone is not a bad thing. And they have value here and very little froth left to shed. And most importantly, they still provide excellent products.So I can buy NVDA stock for the long-term, and I don't worry about the short-term dips for as long as my assumptions remain true. For those who prefer to trade around the short-term levels, there are some lines to know.Above $147, it could target $156 per share. But there are resistance levels in the way. On June 5, there was a sharp reversal from $146.20, so I have to mark it as important as well. Conversely, if the bears can break through $139.75, they could get the chance to set a new low below $132.There are outside factors too. It is important to note that any rallies in Nvidia stock will need the help of the global equity markets. We are still under the threat of geopolitical headlines because of the economic wars that are going on, so it would be smart to take positions in tranches and not all at once. This would leave room to manage dips.Nicolas Chahine is the managing director of SellSpreads.com. As of this writing, he did not hold a position in any of the aforementioned securities. Join his live chat room free here. More From InvestorPlace * 4 Top American Penny Pot Stocks (Buy Before June 21) * 7 S&P 500 Dividend Stocks to Buy at Least Yielding 3% * 7 Stocks to Buy That Don't Care About Tariffs * 5 Healthcare Stocks to Pick Up From the Wreckage Compare Brokers The post Nvidia Stock Is Out of Favor on Wall Street -- Own It Anyway appeared first on InvestorPlace.
Joining Yahoo Finance's Jen Rogers and Myles Udland is Jared Blikre to break down the week's market action in the S&P 500, its 11 sectors (where tech is leading the year again), as well as the weekly winners in the Nasdaq 100 — all with the help of our new YFi Interactive touch screen.

domain: economics


text:
Elevated blood cholesterol is not good. It contributes, for example, to myocardial infarction, atherosclerosis of blood vessels or coronary diseases. Phytosterols are an effective way to avoid these types of problems and to effectively lower cholesterol in the body. Their source is, among others vegetables, oils and nuts. How exactly do they work? How to use them properly? What doses will be the best? Phytosterols and cholesterol - what should we know?
Cholesterol is produced by humans and animals. In turn, phytosterols - meaning plant sterols - can only be produced by plants. Their structure is very similar to the cholesterol design and helps in its reduction. They consist of two basic groups of sterols and stanols.
Sterols have only unsaturated bonds in the molecule, and stanols have a saturated bond. Their action is based on the inclusion of cholesterol in cell membranes. They manage cavities in its particles, at the same time strengthening the entire membrane. They have a vegetable origin, which is why they are most often supplied in food. Phytosterols began their first research in 1950, when it was noticed that their effect on high cholesterol was decreasing. They quickly found their application in health prophylaxis.
Phytosterols - occurrence
Plant sterols are found primarily in plant cells. Natural products mention below are their main source:
- vegetable oils, i.e. olive oil, sesame oil, corn oil or sunflower oil;
- rice bran;
- sesame, sunflower seeds;
- fruits and vegetables;
- wholegrain cereal products;
- leguminous vegetables, i.e. beans;
On the store shelves you can find many products enriched with plant sterols. Among them, yogurts, mayonnaise, ripening cheese, homogenized cheese, confectionery (chocolate), meat products or milk drinks are worth mentioning.
Phytosterols - dosage
Excess plant sterols bring certain side effects - it is worth paying attention to their amount in the diet. Every day a man consumes an average of 500mg stanol and sterols - this is not enough. Daily meals should provide from 1 to 3g of phytosterols. Such a dose will certainly have a positive effect on the functioning of the body.
How do phytosterols work?
How do phytosterols work? What effects do you bring? Can they also pose a threat to people?
The use of sterols has good sides.
- The main and most important effect of using phytosterols is the lowering of blood cholesterol. Sterols inhibit its absorption in the gastrointestinal tract, increasing its excretion from the body. A very small amount goes into the bloodstream.
- Plant sterols in the company of vitamins C and E inhibit platelet aggregation. So blocking the clumping of tiles, which by such clumping can lead to blockages in the circulatory system.
- Plant sterols have an antioxidant effect, i.e. they weaken free radicals in the body and through apoptosis, they cause cell death. Free radicals in the body cause degradation of the DNA molecule, which often contributes to the formation of tumors. These sterols retain, for example, lung, stomach and colon cancer. They also prevent metastases.
- They are often used in cosmetics because they can activate collagen. Thus, they improve skin tone and hydration, reduce wrinkles, accelerate skin regeneration and soothe numerous irritations.
- In the human body they act antifungal and antibacterial.
- They act anti-inflammatory, because they reduce the strong immune response. Thanks to this, they support and inhibit the development of diseases, i.e. rheumatological arthritis.
- Systematic consumption of plant sterols positively affects the prostate - reducing its hypertrophy and alleviating problems with passing urine.
In that case, can phytosterols pose any risk? It turns out that yes.
Their too high dose causes a bioavailability reduction of:
- fat-soluble vitamins,
Rinsing such important vitamins reduces immunity, which carries a greater risk of disease. Phytosterols should not be given to children, because at their young age their body needs cholesterol (as a building block).
Phytosterols and cholesterol
Plant sterols are a natural remedy for many diseases. The relationship phytosterols and cholesterol is very important - their main task is to lower the latter. However, they can also positively affect the skin, or strengthen other systems in the body. Do not exceed the daily dose, because excess amounts of valuable vitamins from the body. This in turn significantly affects the body's immunity. Adequate supply of phytosterols in the diet is certainly a better choice than a series of tablets and medicines.
Where to get phytosterols?
There is no problem with buying them - they are a component of many products that are on store shelves.

domain: medical


text:
Deciding what to plant in May can be difficult – but for all the right reasons because now the weather is improving it gives you so much choice!
The end of the month should be reliably warm so most things can be planted out by then, but at the start of May we still have to be cautious of nocturnal frosts in some colder climates, so keep fleece to hand to throw over plants when very cold nights are forecast.
Most seeds can be sown directly into the soil now and perennials can be planted, though do remember to water regularly should we – and we can but dream – have a warm, dry summer.
So make finalizing your choice of what to plant when easier with our round-up of suggestions for your planting list for the month ahead.
10 suggestions for what to plant in May
By the time May draws to an end, temperatures should be reliably higher, meaning we should be able to plant what we want.
Unless you live in northern climates or have a very exposed garden, you can start planting out tender bedding and less hardy perennials that were brought indoors for the winter, though in cooler areas it’s best to wait until the tail-end of the month.
Make sure you are hardening off plants properly before putting them out as they need enough time to toughen up so they are ready to face the uncertain weathers of the outside garden.
1. Helenium or 'sneezeweeds'
When we think about what to plant in May we often think about the high summer months when we want the garden to be at its peak. But we should also be thinking about the tail-end of summer and fall, when many plants start to fade and the garden runs the risk of looking a little sad.
Filling this colour gap isn’t difficult, as all dahlias will be in their prime and with the right care, summer bedding will still have plenty of oomph left.
One variety that I think is a must for your autumnal patch, however, is Heleniums, also delightfully known as sneezeweeds. They produce daisy-like flowers with hearty centers in burnished shades of gold, orange, yellow and red, make excellent cutting garden flowers and are a perfect end-of-season food source for pollinators.
For the best results, their favourite soil type is fertile and free-draining in a sunny spot. They also need watering and feeding regularly for the best results. Heleniums are an easy perennial plant to propagate by dividing, so do this every few years when clumps get too big.
Top tip: Heleniums look glorious planted in a ‘prairie style’ border with ornamental grasses, echinaceas and red hot pokers.
If your garden has shady areas that need illuminating, why not pop in a hosta or two? Yes, they can be a magnet for slugs and snails (and we have plenty of eco advice on how to get rid of slugs to help you keep your plants safe) but they are also beautiful, their leaves either plain green, ribbed, variegated or blotched with grey-blues and acid greens.
For once, the flowers are not the plants’ main selling point, but when they do emerge they appear as eye-catching mauve or white trumpets. Hosta flowers are often sweetly scented too, though not usually regarded as one of the best fragrant plants available.
Size is no problem, as miniature varieties such as ‘Blue Mouse Ears’ or ‘Pandoras Box’ grow well in garden planters and if you do have a serious slug problem, try a resistant variety – ‘All that Jazz, ‘Big Momma’ and ‘August Moon’ are three that have no truck with molluscs!
Hostas are one of the most popular shade-loving plants, and will do well when planted under trees. They like soil that has been enriched with mulch and do remember to keep them well-watered. They are best avoided if you have thin sandy soil and an exposed garden.
Top tip: Hostas are fully hardy but die right back in winter, emerging again in spring. Large clumps can be divided in autumn or spring.
3. Passion flower
If you want a lot of bang for your buck when thinking about what to plant in May, why not try growing a passion flower to add interest and cover a fence or wall?
We used to have one climbing up the front of our previous house and it was a glorious plant, producing exotic flowers, named after the passion of Christ on the cross, in summer followed by edible, but not particularly delicious, orange fruits in autumn.
Passion flowers are one of the most popular climbing plants. They are relatively low maintenance, and can be safely grown up walls as their roots are not far-reaching and won’t leach moisture from the foundations.
They are easy to care for – simply give a light prune after flowering and feed with a granular fertilizer in spring and mulch well in autumn. However, they are not reliably winter hardy so for the best results plant them in a sheltered site that gets lots of sun.
Top tip: Passion flowers grown from seed can take up to 10 years to flower so if you want to propagate, take cuttings from plants instead.
Sometimes it is easy to find the incessant flowers of summer bedding a little bit sickly sweet, so one solution is to break it up by planting some coleus.
Also known as solenostemon or painted nettle plants, they are one of the best foliage plants to add to the garden thanks to their leaves that come in astoundingly mixed shades, ranging from the acidic yellowy greens to deepest burgundy.
A member of the mint family, they make interesting plants for container gardening ideas, they are worth adding to your exotic and tropical planting scheme ideas and can also be grown as indoor plants.
Coleus are not hardy so need to be sown and grown undercover. However, they are perennials – though usually treated as annuals – and can be successfully overwintered indoors.
Once they have been hardened off in summer, set them in a sunny spot, or somewhere with light shade, with fertile soil.
Top tip: Once in growth, pinch out the central tip to encourage bushy growth and fuller leaves.
5. Scented plants
With sultry summer evenings just around the corner, your list of what to plant in May should really include adding some nocturnal scent to your garden.
Evening-scented plants release their perfume as dusk falls as they are designed to be pollinated by nocturnal insects. Many of them are also pale in color so they show up well under garden lights.
They include the pink biennial sweet rocket that produces large clusters of pink flowers, night phlox and border phlox and night-scented stock. Lilies are also renowned for their perfume and the white trumpets of the regal lily (Lilium regale) produce their heady scent after dusk.
Grow them close to areas where you sit at night and enjoy their delicious scents.
Top tip: Night phlox is not reliably hardy so is best grown in containers so it can be moved undercover in winter.
Fuchsias never go out of fashion and it's easy to see why. There are so many varieties, with jolly skirt-shaped flowers in a multitude of shapes and shades.
If you want pared-back and simple, clean lines, then there’s a fuchsia for you to plant in May. If you are after something much more flouncy and eye-catching then yes, there’s something for you too!
Fuchsias originate in South America and while many are hardy, a lot of those sold as summer bedding won’t tolerate cold winters so should be either grown in pots so they can be moved somewhere frost-free in autumn, or grown as annuals and discarded after summer.
You can also get trailing varieties that are one of the best plants to include in your hanging basket ideas and all varieties sit well in summer patio pots.
If you are wondering how to grow fuchsias, they all appreciate a sunny spot, or somewhere with light shade, and free-draining, fertile soil. Avoid places with full sun in the height of summer as they can cause scorching, and grow them out of windy sites as you risk losing all the flowers.
Top tip: Wait until the end of the month when the risk of frost has passed before planting out half-hardy fuchsias.
7. Biennials such as foxgloves, aquilegia and wallflowers
Biennials are plants that are sown and grown one year, before they go dormant in winter then spring back to life in their second year to flower, set seed and die.
They include honesty, sweet rocket, foxgloves, night scented stock, aquilegia, vipers bugloss and wallflowers.
Factor these beautiful plants into your flower bed ideas and plant the seeds now undercover, growing them in your greenhouse or cold frame. They will then be ready to plant out in late summer or early fall when the soil is still warm, and will give you a splendid show next year.
Nigella is a hardy annual that is often sown directly into the soil in autumn and treated as a biennial. Most others give the best results when sown in trays of seed compost and nurtured in the greenhouse until they are large enough to plant out.
Top tip: Always sow honesty directly where you want it to grow as the seedlings quickly develop deep tap roots and don’t transplant well.
Once seen as a bit ‘fuddy duddy’ and matronly, lavender is very much back en vogue – and rightly so. These easy-going plants fit perfectly into a Mediterranean garden scheme and are incredibly versatile and low-maintenance, flowering in a variety of shades from darkest blue to white, all offset by attractive grey-green leaves.
They are tough plants too, happy in poor soil and therefore ideal for less-able gardeners who may have trouble with watering.
If you're thinking of landscaping with lavender, it looks stunning in gravel gardens, but not out of place in traditional cottage gardens.
The two main varieties that are ideal to plant in May are English, with masses of small flower heads, and French, which has larger flowers topped by a tuft of petals.
They like free-draining soil and the French varieties are best planted in containers so they can be moved undercover in winter as they are less hardy then their English cousins.
Top tip: Lavender is one of the many garden plants that are great for drying, either for use in an arrangement or to scent drawers and clothes. Cut stems just before the flowers are fully open, then hang them upside down in bunches. When completely dry, either rub the stems between your hands to loosen the flowers or keep them intact to use in flower arrangements.
Who doesn’t love a patch of pellies in their garden? These ubiquitous summer bedders that are as suitable for planter box ideas and hanging baskets as they are for adding to garden border planting ideas, come in a wide range of colors, from white to scarlet and all shades of pink in between.
Some have scented leaves, others have patterned foliage and they all make a fabulous summer splash in borders and containers.
Although they are tender they are worth considering when thinking about what to plant in May, though in colder areas I would wait until the end of the month, because they will still turn up their toes after a late frost.
Most varieties love full sun, though Regal and Zonal cultivars prefer some shade. Pelargoniums can survive with little watering – certainly don’t overwater any growing in pots – but they will thank you for a regular feed with a high-potassium liquid tomato fertilizer to keep the flowers coming.
Pinch out the shoot tips after planting to encourage bushy growth and make sure they have good ventilation as the fleshy leaves are vulnerable to mildew and tots.
Top tip: Take softwood cuttings in late summer in case the parent plants don’t make it through the winter.
If you need something speedy to sow in time for a special summer event, or just to fill some border gaps, low-maintenance annuals are your go-to option.
Sown now, when the soil is warm and damp, they will germinate and hopefully even flower by midsummer. Sunny calendulas, nigella with its host of colors and elegant, delicate foliage, cornflowers, annual poppies, and cheery California poppies can all be started now to flower this summer and fall.
To grow flowers from seeds, prepare a seedbed by raking the soil to break up large lumps, then dampen with fresh water before scattering the seeds as thinly as possible.
Cover with a little more soil, label the area and puff some pepper dust over the top to keep off pests and cats wanting to use it as a litter tray. If seedlings appear too close together you can thin them by hand.
Top tip: Many hardy annuals self-seed freely so remember to collect some seeds in autumn to sow where you want them to grow.
Ruth is the gardening editor of Amateur Gardening magazine and spends her working days carrying out, writing about and photographing the tasks the readers should be carrying out each week, as well as testing many of the new products that arrive on the gardening market. She is horticulturally trained, with a qualification from the Royal Horticultural Society.
Take part in the RSPB’s Big Garden Birdwatch 2023 to save our feathered friends
Gardens Watching garden visitors for just one hour in the Big Garden Birdwatch 2023 could help provide vital data to protect birds from the effects of climate change
By Jayne Dowle Published
Do you need to chit potatoes? Find out what the experts say
Grow Your Own Learn how to chit potatoes before planting them in the ground and you’ll be on your way to getting an earlier and bigger harvest
By Drew Swainston Published

domain: agronomy


MusAic for Nokia 5300, 7373, and 7390 1.2.3
Requirements: Symbian, J2ME MIDP 2.0, CLDC 1.1
Limitations: Some features disabled
Publisher's description of MusAic for Nokia 5300, 7373, and 7390
MusAic creates polyphonic music in any genre, using algorithms from the field of Artificial Intelligence. Use as a never ending jukebox. Create your own, absolutely unique ringtones with just a few simple clicks. Generate ideas for fast and creative music composing. Each new song is completely original, created as a result of your rating input and the programs reaction to it. The program adapts to your musical tastes, and this amounted knowledge can be saved as a Virtual Composer. You can even "Marry" two composers and find out what their offsprings music sounds like. All songs can be saved and exported as a MIDI file to your phones "Sounds" directory. From there, it can be set as a ringtone or transferred to a computer and opened in a Sequencer program for editing. This way, MusAic works as an idea generator for fast and creative music composing.
Click here to download
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TULSA — A child advocacy group asked Monday for $9.5 million for the legal effort its attorneys and others put into a federal class-action lawsuit that pushed DHS into agreeing to reforms.
Children's Rights, a New York-based group, asked a federal judge for $8,345,588 for its attorneys' time on the case and $912,711 for the group's legal expenses.
The group asked that a New York firm that helped in the case be paid $262,120 for the firm's expenses.
The advocacy group's executive director, Marcia Lowry, is seeking to be paid at a New York City rate of $700 an hour. Other attorneys are seeking to be paid from $175 to $375 an hour.
The attorneys argued their request is reasonable and that their dedicated work achieved a historic and life-altering result for Oklahoma's children.
“But for the dedication and resources of the lawyers of Children's Rights, none of these changes would have occurred,” attorneys told U.S. District Judge Gregory Frizzell in their application for payment. “This historic and comprehensive reform did not come easily. The defendants employed the full resources of the state to oppose the reformation.”
Tulsa attorney Frederic Dorwart, who also helped Children's Rights in the case, did not charge for any of his firm's time or expenses.
Officials at the Department of Human Services have budgeted only $4 million to pay the group.
Children's Rights sued DHS officials in 2008 in federal court in Tulsa. The group sued on behalf of the state's foster children.
The group alleged in the lawsuit that DHS policies and practices are so bad that neglected and abused children are being harmed or are at risk of harm at state shelters and foster homes.
DHS commissioners agreed to make reforms to its child-welfare operations when they voted in January to settle the lawsuit.
Commissioners settled rather than risk losing at a trial and having the judge possibly order even more costly reforms. DHS is calling the improvements “The Oklahoma Pinnacle Plan.”
The plan's cost
DHS plans to hire more child-welfare workers, recruit more foster homes and phase out using shelters to care for the youngest children. Officials initially estimated DHS will need almost $100 million more in state funds a year once all the reforms are in place.
The state already has paid more than $7.6 million in legal fees and expenses to attorneys hired to represent DHS officials in the lawsuit.
The bulk of that money — almost $6.9 million — went to the law firm hired to fight the lawsuit.
Almost $700,000 more went to the attorney who represented DHS officials in the settlement negotiations.
Children's Rights has filed similar class-action lawsuits across the country, including in Texas, Kansas and Missouri.
Children's Rights contends its bill would not be nearly as high if DHS had not engaged in what it contended were “overly aggressive tactics ... motivated by patently unjustified denial of the serious, systemic problems plaguing DHS.”
DHS is expected to ask the judge to pay Children's Rights much less than it is asking.
“I believe we'll definitely question the fees,” said Richard DeVaughn, one of three DHS commissioners who voted against the settlement.
“It's about the money. It wasn't about the children,” DeVaughn said of the group's lawsuit.
“The reason they were so willing to settle at the end was we had them on the ropes.”


text:
Carlo Gesualdo (1566 – 1613) was an Italian composer of the late Renaissance era. His father was prince of Venosa, and his mother was the niece of then-current pope Pius IV and the sister of Cardinal Borromo, who would later be canonized as a saint in the Catholic Church. From an early age, Carlo showed an interest in writing and playing music. He learned to play several instruments but his favorite was the lute. As a composer he is best known for writing expressive madrigals and pieces of sacred music.
His music is among the most experimental and expressive of the Renaissance, and without question is the most wildly chromatic. Progressions such as those written by Gesualdo did not appear again in music until the 19th century, and then in a context of tonality. Although widely admired as a composer, the best known fact of his life is his brutal and violent killing of his first wife and her aristocratic lover when he caught them in the act of infidelity. As a nobleman, Gesualdo was not formally charged with a crime. However, his high societal rank provided virtually no protection from potentially vengeful relatives of the deceased. His former wife and her lover had belonged to two of the most powerful and wealthy families in southern Italy, and so Gesualdo immediately made provisions to fortify his castle and hired guards to protect himself. This refuge became his permanent residence.
The evidence that Gesualdo was tortured by guilt for the remainder of his life is considerable, and he may have given expression to it in his music. Simone Molinaro (a contemporary of Gesualdo and also featuring in this website) greatly admired Gesualdo. In 1609 Molinaro met Gesualdo in Naples. Two years later Molinaro published Gesualdo’s six-volume collection of madrigals for five voices. The publication of the Gesualdo madrigals was ground-breaking because it presented Gesualdo’s music in full score as opposed to partbook format.
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domain: artistic


text:
Scholastic Inc | ISBN 9780545833219 Hardcover
32 Pages | 11.79" x 9.74" | Ages 3 to 5
Scholastic Inc | ISBN 9780545913775 Ebook
32 Pages | Ages 3 to 5
Get transported to a lush, vibrant jungle in this gorgeously illustrated picture book about facing your fears.
Richard hears something in his room before bedtime. Is it a monster? He doesn’t wait to find out and sets off running through the streets, over the hills, through the forest, and into the fields until he finds himself in a magical jungle. With the help of his stuffed lion Lionheart, Richard finds the courage he needs to face his fears.
Raves & reviews:
Praise for When It Snows
“ In his first picture book... Richard Collingridge dives headlong into a fantasy of the season, showing it to be a vast and mountainous expanse of white, both eerie and enchanting.” —The New York Times
“ Newcomer Collingridge’s paintings are sumptuous and absorbing, creating a genuine sense of magic.” —Publishers Weekly


text:
You already gave your best. You can't change the results now. What you can do is prepare for an upcoming exam in a planned manner. there is no use pondering over what has passed and what is no longer in your control. Just let it go. Who know, you might actually have done well in the exam. Don't overthink it
that's what I am trying to do but somehow I end up thinking about it.
Keep yourself occupied in other stuff .. hang out with friends, listen to music, or do anything that you like.. so that your mind doesn't wander off

domain: psychology


text:
It’s not much to look at, but researchers at the University of Maryland have created an amazing “Robo Raven” that almost perfectly mimics the flight of real birds.
Lots of people have tried to build something like this, but it never looks quite right. The trick is that Robo Raven’s wings move independently of one another and can be programmed to perform any desired motion. It climbs, glides, dives and turns almost as gracefully as the real thing — so real that even a hawk is fooled into attacking it.
Developed by University of Maryland robotics professors S.K. Gupta and Hugh Bruck, the Robo Raven has two programmable motors that synchronize electronically to coordinate motion between the wings, allowing for complex aerobatic manoeuvres. What application this could possible have is not yet clear but it’s still very impressive.

domain: news


The present study aims at developing a surrogate modeling method able to represent both non-stationarity and the propagation of uncertainties in the dynamic response of complex structures with time-dependent characteristics. Toward this end, AutoRegressive (ARX) models with random time-varying parameters are introduced. More specifically, the parameters of the introduced model are expanded on a multidimensional basis, with one of the dimensions corresponding to a scheduling variable dependent of time and the rest of them to the probability space of the uncertain input parameters. Linear parameter varying basis functions and polynomial chaos basis are utilized for this purpose with the resulting Polynomial Chaos Linear Parameter Varying ARX (PC-LPV-ARX) model being fully described by a finite number of deterministic coefficients of projection that may be estimated by a linear maximum likelihood method. In order to illustrate the workings of the method, the whole procedure is applied for the construction of a surrogate model of the vibration response of a wind turbine blade under normal operation. The surrogate is estimated from simulated data obtained by FAST (an aeroelastic computeraided engineering tool for the simulation of horizontal axis wind turbines), while the value of the average wind speed within each analysis period is assumed to be uncertain following a normal distribution. In overall, thisstudy aims at demonstrating the effectiveness and applicability of the proposed method for the estimation of non-stationary surrogate models of low order that are capable of accurate approximation of large scale numerical models.
|Titel||Safety and Reliability of Complex Engineered Systems - Proceedings of the 25th European Safety and Reliability Conference, ESREL 2015|
|Redaktører||Luca Podofillini, Bruno Sudret, Božidar Stojadinović, Enrico Zio, Wolfgang Kröger|
|Status||Udgivet - 2015|


text:
“Moby Duck,” which was published by Viking, an imprint of Penguin Random House, lends itself ideally to the IMAX experience by drawing from the rich IMAX heritage of 3D adventure documentaries. As a film, Moby Duck will dramatize such interlocking threads as oceanic currents, the effects of plastics on wildlife, and the phenomenon of gyres and the Great Pacific Garbage Patch.
“This is an exciting new project for IMAX Entertainment, one that goes hand-in-hand with our legacy but with a fun twist,” said Greg Foster, Senior Executive Vice President, IMAX Corporation and CEO of IMAX Entertainment. “By optioning rights to ‘Moby Duck,’ a thrilling and entertaining global journey based on an unbelievable true story, IMAX continues to provide quality entertainment to our fans around the world. We are thrilled to work again with our partner Drew Fellman and look forward to this out-of-the-box addition to IMAX’s decades-long success of documentary filmmaking.”
“From the very first pages of ‘Moby Duck,’ I knew this riveting tale would make an amazing IMAX film that would be wildly entertaining and also tackle vital environmental issues,” said Fellman. “This story of thousands of toys lost at sea illustrates how something as simple as a rubber ducky can reveal natural mysteries as deep as the ocean itself.”
The IMAX 3D release of Moby Duck will be captured with IMAX cameras. The crystal-clear images, coupled with IMAX’s customized theatre geometry and powerful digital audio, create a unique environment that will make audiences feel as if they are in the movie.
When the writer Donovan Hohn heard of the mysterious loss of thousands of bath toys at sea, he figured he would interview a few oceanographers, talk to a few beachcombers, and read up on Arctic science and geography. But questions can be like ocean currents: wade in too far, and they carry you away. Hohn’s accidental odyssey pulls him into the secretive arena of shipping conglomerates, the daring work of Arctic researchers, the lunatic risks of maverick sailors, and the shadowy world of Chinese toy factories. Moby Duck is a journey into the heart of the sea and an adventure through science, myth, the global economy, and some of the worst weather imaginable.

domain: geography


text:
The Bear and the Nightingale
A NovelBook - 2017
From Library Staff
Arden's debut takes place in a fantasy old-world Russia, where folk tales and magic coexist with reality. Daughter of a prosperous landowner, Vasya has always been different. Like her late mother, she can perceive creatures that others can't, such as the benevolent guardians of the hearth and sta... Read More »
Chicago Public Library recommends this title as one of the best books published in 2017.
From the critics
AgeAdd Age Suitability
ArapahoeKatieK thinks this title is suitable for 12 years and over
SummaryAdd a Summary
A story about Russian folklore. A girl is born to a Russian landowner in the middle of winter and there is something special about her. A tomboy from an early age, she spends her first decade and a half figuring out who she really is and how to use her abilities, how to fight evil, and how to keep her family safe.
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text:
Search the Community
Showing results for tags 'hyperram'.
Hi all and Digilent representatives! I am a "long" user of the Cmod A7 and am quite pleased with it, except for the choice of SRAM. And on the side I have been playing with another RAM that is easy to use, delivers better performance (about 200 MB/s for the 3.3v version), and more space (8Mbyes) which would be a really nice update to the Cmod-A7. The new "HyperRAM" (http://www.digikey.com/products/en/integrated-circuits-ics/memory/774?k=hyperram&k=&pkeyword=hyperram&pv142=133&FV=ffe00306%2C1c0011&mnonly=0&newproducts=0&ColumnSort=0&page=1&stock=1&am

domain: hobby


text:
Camila Cabello, Shawn Mendes Release New Song Together
Cabello and Mendes finally release collaboration video
Lahore (21st June, 2019): “If I Can’t Have You” singer, Shawn Mendes and Camila Cabello release first collaboration since 2015.
The “Havena” singer has finally released a new song in collaboration with friend and singer, Shawn Mendes. This is a follow up of the 2015 hit “I Know What You Did Last Summer”. According to Baaghi TV reports, this new release is a bittersweet ballad of love titled, “Señorita”.
As bittersweet as can be, the video presents Cabello as a waitress who becomes involved with a customer, portrayed by Mendes in all his guitar-slinging glory. Shown to be passionately in love with each other, the plot slowly unfolds to the idea that the lovers are not meant to be together.
Shawn Mendes who has previously produced hit songs has recently released his first non-album single titled, “If I Can’t Have You”. He is currently in the middle of his North American summer tour, as confrmed by Rolling Stones.
While on the other hand, Camila Cabello is super busy collaborating with other artists including Mark Ronson, Cardi B and even Ed Sheeran.
Stay tuned to Baaghi TV for more updates.

domain: celebrity


LIAM ICON JOS 424 50SPS SKINNY FIT JEANS
JACK&JONES JEANS INTELLIGENCE
- Skinny fit jeans for a rough rock ‘n’ roll look
- Five-pocket style with a modern look
- Stretchy denim for maximum comfort
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- The model is wearing a size 31/32 and is 187 cm tall
- JACK & JONES JEANS INTELLIGENCE
Skinny fit Liam
Liam is the skinniest fit we’ve got. Liam gives you that sharp, super tight silhouette that comes with a skinny fit. Of course, we’ve made sure that Liam stretches in all the right places.
Icon is a modern interpretation of the classic five-pocket jean. It has all the familiar details, but with a contemporary twist. It’s a classic, just a little more up-to-date but without too many bells and whistles.
Super Stretch 50%
Super Stretch 50% is denim that stretches 1.5x its actual size. The high amount of stretch makes the denim perfect for skin-tight jeans. And you don’t have to worry about jeans that get saggy once you’ve worn them a few times. The special yarn spun from a unique mix of cotton, polyester and elastane gets rid of that concern.
- 92% Cotton, 6% Polyester, 2% Elastane
Machine wash at max 40°C under gentle wash programme
Do not bleach
Do not tumble dry
Low temp. iron. Highest temp. 100°C
Dry clean (no trichloroethylene)
- Article Number: 12170392


text:
At a November 23 briefing, key New Hampshire State agencies provided updated revenue estimates to policymakers that revealed a much more optimistic revenue outlook than previous projections. With tax receipts performing well in the State fiscal year thus far, and agencies substantially underspending their budgets last year, the total budget shortfall may be much more manageable and put fewer programs at risk than previously anticipated. Policymakers may be able to offset much of any deficit with the Rainy Day Fund and support or expand programs and services for Granite Staters, who have seen their health and financial security put at risk during the pandemic.
The State agencies projected an aggregate revenue shortfall in the General and Education Trust Funds of between $75.9 million (2.9 percent) and $3.7 million (0.1 percent) below the amount of revenue planned for the State Budget in State Fiscal Year (SFY) 2021. This compares to the approximately $239 million projected shortfall calculated in September, and the draft audited revenue shortfall figure of $106 million for SFY 2020. The change in estimates occurs as revenues to the General and Education Trust Funds are $15.6 million (2.5 percent) ahead of the State Revenue Plan as of the end of October even after anomalous payments are removed, and early receipts for November appear favorable as well. Other states have also seen revenue shortfalls smaller than initially expected.
The primary revisions to New Hampshire’s revenue forecast result from upward revisions to five tax revenue sources: the Business Profits Tax (BPT), the Business Enterprise Tax (BET), the Tobacco Tax, the Interest and Dividends (I&D) Tax, and the Real Estate Transfer Tax (RETT). The BPT and BET, which are typically paid together by businesses, may still end the year lower than planned, but revenue projections were revised substantially upward. The New Hampshire Department of Revenue Administration (DRA) noted that the upward revision was prompted by the strength of payments from some of the largest businesses, suggesting these companies are remaining profitable. The DRA indicated the medical and technological industries, as well as home improvement and box stores, were performing well. Unlike a traditional recession, when nearly all activity slows, tax receipts suggest some businesses have seen significant revenue losses while others had substantial increases in business.
Tobacco Tax revenues have remained elevated, which appeared to be initially due to wholesalers stocking up on tax stamps. However, a ban on mentholated tobacco and other flavored tobacco products in Massachusetts, which began in June, may have shifted some purchases to New Hampshire. Also, the stresses on individuals caused by the COVID-19 pandemic may be prompting greater use among people who would otherwise smoke less or quit smoking.
Revenues from the I&D Tax continue to be buoyed by the strong performance of the stock market, as well as savings from those earning interest on assets such as bank accounts. RETT revenues have been supported by increases in real estate sales and home prices. The number of taxable RETT transactions was up 17.5 percent in September activity relative to last year, and prices were up 32.1 percent, likely putting home purchases out of reach of many Granite Staters.
While revenues to the General and Education Trust Funds have improved, a significant deficit is still projected for the Highway Fund. This shortfall is primarily driven by lower Motor Fuels Tax receipts, which were reported as $10.4 million (8.0 percent) lower than planned in SFY 2020 and are projected to be $15.2 million (11.5 percent) short in SFY 2021. While the Highway Fund is a separate fund, General Fund surplus has been used to support the Highway Fund in the last two State Budgets.
The State’s fiscal situation is substantially better than suggested by the early indications of the pandemic’s impacts. In SFY 2020, State agencies underspent their budget allocations from the General Fund; this underspending, combined with the revenue shortfall, resulted in a preliminary budget deficit of $67 million for SFY 2020. This SFY 2020 deficit, combined with the new revenue projections for SFY 2021, suggest the Rainy Day Fund’s $115.5 million could substantially offset the budget shortfall. Policymakers have the tools and resources available to protect State services at a time when more Granite Staters are in need.
Although recent revenue estimates are less dire than earlier iterations, these projections are subject to change, and the future of the pandemic is not yet clear. If substantial limits on activity are put in place to protect public health again, tax revenues may fall closer to the levels feared previously. Additionally, a general economic slowdown resulting from widespread income losses could limit revenues in both this State Budget and the next one. Careful planning by policymakers will be required to ensure needed services are funded, economic growth is supported, and Granite Staters can recover from the COVID-19 crisis.
– Phil Sletten, Senior Policy Analyst

domain: finance


text:
The Wire Troll: Kyle Orton Making the Cowboys Jealous
Zach Mettenberger is the latest rookie QB to thrust himself into the spotlight. (George Walker IV/The Tennessean)
The mistakes we make are far more trivial than the life we lead outside the Fantasy realm. Yet, with sword unsheathed and no shield to be found we battle on through nightmarish Sundays and micro managing Mondays. We know the stress you feel is probably as tangled as the metaphors we use, so relax in your favourite chair and read our Wire Troll picks for Week 10.
Bye Weeks: Indianapolis, Minnesota, Washington, Houston, New England, San Diego
Zach Mettenberger, Tennessee Titans: What’s this? Another rookie QB? It seems Mettenberger got more press for his selfies than he did for his nearly 300 passing yards vs. the Texans in his first career start. We know the Titans are not a great team, but what they are is a bunch of cats with enough experience to make them potentially dangerous. They do a great job of spreading the ball around and while that may suck for owners of guys like Bishop Sankey and Justin Hunter, it makes their trigger man Mettenberger alluring. You better learn how to pronounce this young QB’s name. Need help? Check the video below.
Kyle Orton, Buffalo Bills: In his last start before the bye, Orton was 10-for-17 passing. Not impressed, are you? Well, he also threw four TDs passes. The Bills were playing the Jets, but consider the experience level and maturity Orton has and you will realize no other free agent in your league can bring that to the table.
Terrance West, Cleveland Browns: So there’s this movie called Cop Out with Bruce Willis and Tracy Morgan and it’s not very good. Somehow it’s more laughable than the Browns RBBC trio. For some reason Cleveland decided to give West the bulk of carries versus the vaunted (clearly joking) Tampa Defense. Isaiah Crowell did not get anywhere near the pigskin and Ben Tate was largely ineffective. So basically they are roster-worthy players to start in a pinch. West is owned the least, so snag him with the same nails you are using to try to claw your way up through the standings with, as Cleveland is apparently taking turns spreading the wealth of touches around.
Stevie Johnson, San Francisco 49ers: Do we really need to sell you on Johnson? You know he is skilled and Anquan Boldin as well as Michael Crabtree just aid his openness. Trading for Johnson was a win now move for the 49ers and now they are .500 team. Look for an all hands on deck situation moving forward and even though Johnson might be the third or fourth wheel, he will still be needed.
Andre Roberts, Washington Redskins: Among a gaggle of speedy flashes like WR DeSean Jackson and powerful RB Alfred Morris, stands a 5’11” pillar of grit named Andre Roberts. When the Redskins signed Roberts last spring, it wasn’t to be their Megatron. Roberts has played second or third fiddle before. But, trust us we say he is still a bow-worthy music maker.
Andrew Quarless, Green Bay Packers: Why in the name of anti-freezing tundra coils are we recommending Quarless? He has not received more than five targets or gained more than 43 yards in any one game this season. He does have two scores on the year and last season when Jermichael Finley almost died on the field, Quarless became a contributing cog. The Pack brought in bodies this offseason to push Quarless and he has put his head down and worked hard. Aaron Rodgers trusts the veteran TE and so should you as this final week of byes ticks away.
RotoRob’s Fantasy Football Weekly Podcast
Crave more Fantasy football chatter? Then join us every Wednesday at 1:30 p.m. EST for RotoRob Fantasy Football Weekly Podcast on Blogtalkradio as we commit personal fouls and chop-blocks for 90 minutes or so each week. Tune in here.

domain: sports


text:
Pickton, North Riding of Yorkshire
Pickton, a township in Kirk Leavington parish, in the N.R. Yorkshire, 4 miles S of Yarm. It has a station on the N.E.R. Post town and money order and telegraph office, Yarm. Acreage, 1003; population, 114. There is a Wesleyan chapel.
Land and Property
The Return of Owners of Land in 1873 for the North Riding of Yorkshire is available to browse.
Newspapers and Periodicals
The British Newspaper Archive have fully searchable digitised copies of the following North Riding newspapers online:

domain: history


MrOwl has multiple immediate openings for Software Developer, Applications 1 in Albuquerque, NM. These positions for Software Developer, Applications 1 will play a key role in assisting MrOwl in building a new technology using Java, C, & C++ in a Linux environment that will change the internet forever. Specific duties include using object-oriented programming to build a cloud computing platform utilizing databases; designing database structures; managing databases and using data mining to put information into databases; designing and maintaining cloud computing platform security; and using machine learning and knowledge of neural networks to write software to learn patterns from existing data and adapt the technology accordingly. These positions require a U.S. Master’s degree in Computer Science or Computer Engineering with demonstrable knowledge of C, C++, Java development experience in a Linux environment, cloud computing, neural networks, data warehouse and data mining, network security, object-oriented programming, database management; data structure, and machine learning. Send resume by mail with contact information to Ms. Michaeli Marler, Office/HR Manager, MrOwl.com Inc., 6501 Americas Pkwy NE, Suite 800, Albuquerque, NM 87110.
MrOwl has immediate openings for Software Engineers in Albuquerque, NM. The Software Engineers will write software to create search technologies; retrieve and index information from web sites, documents, images and videos to add into Lucene indexes; adapt search technology to include pattern recognition and statistical learning; and help create a search advertising platform to match advertisements to the user in the search results pages. These positions require a U.S. Master’s degree in Computer Science with demonstrable knowledge of: C, C++, ASP.NET, Java, and Linux scripts; programming, information retrieval, mining and integration (search engines) in Lucene; advanced cryptographic algorithms and cryptographic techniques; advanced data mining techniques, i.e., classification, clustering and mining text and web data; search technology including inverted indexes in using Lucene; search advertising; and machine learning and neural networks. Send resume by mail with contact information to Ms. Michaeli Marler, Office/HR Manager, MrOwl.com Inc., 6501 Americas Pkwy NE, Suite 800, Albuquerque, NM 87110.