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Hereditary canine spinal muscular atrophy (HCSMA) is an autosomal dominant degenerative disorder of motor neurons. In homozygous animals, motor units produce decreased force output and fail during repetitive activity. Previous studies suggest that decreased efficacy of neuromuscular transmission underlies these abnormalities. To examine this, we recorded muscle fiber endplate currents (EPCs) and found reduced amplitudes and increased failures during nerve stimulation in homozygotes compared with wild-type controls. Comparison of EPC amplitudes with muscle fiber current thresholds indicate that many EPCs from homozygotes fall below threshold for activating muscle fibers but can be raised above threshold following potentiation. To determine whether axonal abnormalities might play a role in causing motor unit dysfunction, we examined the postnatal maturation of axonal conduction velocity in relation to the appearance of tetanic failure. We also examined intracellularly labeled motor neurons for evidence of axonal neurofilament accumulations, which are found in many instances of motor neuron disease including HCSMA. Despite the appearance of tetanic failure between 90 and 120 days, average motor axon conduction velocity increased with age in homozygotes and achieved adult levels. Normal correlations between motor neuron properties (including conduction velocity) and motor unit properties were also observed. Labeled proximal motor axons of several motor neurons that supplied failing motor units exhibited little or no evidence of axonal swellings. We conclude that decreased release of transmitter from motor terminals underlies motor unit dysfunction in HCSMA and that the mechanisms determining the maturation of axonal conduction velocity and the pattern of correlation between motor neuron and motor unit properties do not contribute to the appearance or evolution of motor unit dysfunction.
The motor neuron diseases are a collection of complex degenerative disorders that have in common motor neuron cell death and degeneration (Mitsumoto et al. 1998;Rowland 1994). Most animal model studies thus focus on understanding the mechanisms that underlie motor neuron cell death and degeneration and define the pathogenesis in terms of these phenomena. Motivating this approach is the hope that, by inhibiting or preventing cell death and/or degeneration, loss of motor unit function will be inhibited and disease progress slowed. In several instances, however, inhibiting cell death in animal models has not halted disease progression (Klivenyi et al. 1999; Li et al. 2000; Sagot et al. 1995, 1996), and effective therapy remains elusive.
In studying a canine model of motor neuron disease [hereditary canine spinal muscular atrophy (HCSMA)], we have used a different approach that focuses instead on defining how motor unit function is initially lost (Pinter et al. 2001a). One reason for this approach is concern that motor neuron cell death and degeneration are endstage phenomena that may not necessarily share mechanisms that provoke loss of motor unit function. An important advantage of our approach is that pathological changes observed in axons, motor terminals, or cell bodies can be related to the causes of weakness (i.e., motor unit dysfunction) to achieve a functionally based understanding of the pathogenesis.
HCSMA is an autosomal dominant, degenerative disorder of motor neurons that shares pathological features with human motor neuron disease (Cork et al. 1982a; Pinter et al. 2001a). Motor unit dysfunction appears to be the most important cause of weakness in HCSMA. In homozygous animals, this dysfunction appears first in slow motor units and manifests as reduced motor unit force output and as an inability to sustain motor unit force output during repetitive activity, termed tetanic failure (Pinter et al. 1995,2001a).
The cellular mechanisms that give rise to motor unit dysfunction in HCSMA are unknown. The dependency of tetanic failure on stimulus frequency and the ability of 4-aminopyridine (4AP) to increase motor unit force output suggest that defective or insufficient release of ACh from motor terminals is involved (Pinter et al. 1997,2001a) but, thus far, direct evidence has not been obtained. We recently found that motor unit tetanic failure in the medial gastrocnemius (MG) muscle of HCSMA homozygotes is not associated with atrophy of the neuromuscular junction (NMJ) or evidence of motor terminal degeneration (Balice-Gordon et al. 2000). This also suggests that motor unit failure may result from functional deficits at the NMJ but it is not clear whether these deficits are localized to the NMJ or are part of a more generalized pattern of motor neuron failure. Evidence of axonal involvement in HCSMA suggests that axonal abnormalities might contribute directly or indirectly to motor unit or neuromuscular transmission failure. Findings include evidence that motor axon size is reduced in HCSMA homozygotes (Cork et al. 1989) and evidence of decreased mRNA for the light neurofilament protein (Muma and Cork 1993). In addition, large accumulations of maloriented neurofilaments are found in some proximal motor axons in HCSMA (Cork et al. 1982b, 1988). Similar aggregations are commonly observed in other forms of motor neuron disease (Hirano 1991; Williamson et al. 1996). These aggregations in particular and abnormalities of the cytoskeletal components of axons in general have received much emphasis as contributing factors mediating the pathogenesis of motor neuron disease (Julien 1995), but when they appear relative to loss of motor unit function is not known.
To gain further insight into the pathogenesis of HCSMA, we examined the status of neuromuscular synaptic transmission in HCSMA homozygotes by recording endplate currents (EPC) in response to nerve stimulation. In this paper, we also investigated axonal function and structural properties during the evolution of motor unit dysfunction. In particular, we studied the postnatal time course of conduction velocity maturation in HCSMA homozygotes in relation to the onset of motor unit dysfunction and the relationships between axonal conduction velocity and motor unit mechanical properties. In addition, we intracellularly labeled several motor neurons in one homozygote after characterizing motor unit dysfunction and examined directly for the presence of swellings in proximal motor axons. Our results demonstrate that EPCs are reduced at homozygote NMJs to the extent that the ability to generate muscle fiber action potentials is likely compromised. This reduction of amplitude is associated with increased EPC failure in response to nerve stimulation. Axonal conduction velocity appears to mature normally during the evolution of motor unit dysfunction, and axonal neurofilament aggregates appear to play no role in either the onset or evolution of motor unit dysfunction in HCSMA. The following paper presents a quantal analysis of neuromuscular transmission in HCSMA homozygotes and provides evidence for a presynaptic deficit of transmitter release at HCSMA homozygote motor terminals (Rich et al. 2002). A portion of these results have been reported in abstract form (Pinter et al. 2001b).
Animals used in this study were obtained from the HCSMA breeding colony or from vendors providing purpose-bred normal animals. All work reported in this paper was approved by Institutional Animal Use and Care Committees.
Homozygous dogs can be identified by 6–8 wk after birth by the appearance of weakness in the tail muscles. In this study, data obtained from homozygotes have been compared with data from adult normal members of the HCSMA pedigree, from symptomless younger animals from the HCSMA colony, and from genetically normal, control animals. We use the term "symptomless" because at ages of less than about 1 yr, HCSMA heterozygotes cannot be readily distinguished from phenotypically normal HCSMA animals. Heterozygotes lack motor symptoms throughout the postnatal interval when homozygotes can be studied (2–6 mo) and begin developing signs about 10 mo after birth (Cork et al. 1982a).
Surgical preparation
One collection of HCSMA homozygotes and symptomless animals from the HCSMA pedigree was used in terminal experiments involving measurement of motor unit force and motor neuron electrical properties or intracellular staining of motor neurons. All dogs were initially anesthetized with 35–40 mg/kg iv sodium pentobarbital. Supplemental doses were administered during the experiment via an intravenous cannula to maintain an absence of withdrawal and corneal reflexes. A tracheal cannula was inserted to maintain a patent airway and to provide for monitoring of end-tidal CO2. Blood pressure was continuously monitored via an arterial cannula. Rectal temperature was monitored and maintained at 37 to 38°C with a heating pad and infrared lamps.
The left MG muscle and its nerve were exposed, and the muscle and its tendon were dissected free of the accompanying lateral gastrocnemius muscle and surrounding connective tissue as much as was safely possible and prepared for attachment to a strain gauge for motor unit force recording. The strain gauge was capable of detecting minimal forces of about 100–200 mg. All force recording was performed using a servo-operated device that maintained muscle preload at 100 g. The MG nerve was freed of surrounding tissue and mounted on a pair of bipolar stimulating/recording electrodes. Fine stainless steel wires (2–4 pairs) were inserted into the MG muscle and connected to high-gain differential amplifiers for recording of motor unit electromyographic (EMG) activity. A laminectomy was performed to expose the lumbosacral spinal cord. The animal was then mounted in a frame that immobilized the vertebral column and the left hindlimb. Exposed tissues were covered with warm mineral oil or Vaseline.
At the conclusion of surgical preparations, all animals received bilateral pneumothorax and were mechanically respirated to minimize respiratory-associated movements during intracellular recording. All animals remained unparalyzed.
Muscle fiber samples for EPC or muscle fiber voltage recording were obtained from a separate group of HCSMA homozygotes and normal animals. For these studies, animals were anesthetized and monitored as described above. Either the left or right MG muscle was freed as completely as possible from surrounding connective tissue so that the muscle could be elevated, allowing an elongated metal support platform to be placed against the deep surface of the muscle. Incisions from the superficial to the deep muscle surface were made orthogonal to the medial–lateral muscle dimension following the paths of tendinous inscriptions visible on the muscle surface. In this way, strips of muscle fibers were obtained that extended the length of the muscle, were about 5 mm in thickness, and contained fibers running uninterrupted between the deep and superficial tendinous surfaces of the muscle. Smaller samples were blocked from these strips and placed into Sylgard-lined dishes through which oxygenated solution flowed. Unless otherwise noted, the composition of the bathing solution was (in mM) 118 NaCl, 3.5 KCl, 2.0 CaCl2, 0.7 MgSO4, 26.2 NaHCO3, 1.7 NaH2PO4, 5.5 glucose (pH 7.3–7.4, bubbled with 95% O2–5% CO2).
Recording and staining procedures
Single MG motor neurons or motor axons were identified by antidromic activation following electrical stimulation of the MG nerve using an intensity 5× threshold for the appearance of whole-muscle EMG. Intracellular recording from ventral root motor axons was performed as described previously (Cope and Clark 1991). Intracellular records were obtained using micropipettes filled with 3 M KCl and connected to a conventional amplifier. Following impalement of an MG motor neuron soma or motor axon, data were collected for measuring motor axon conduction velocity. Conduction velocities were determined by dividing the conduction distance between the spinal cord (or ventral root impalement site) and the peripheral nerve stimulation site (measured postmortem) by the time interval between the peripheral nerve stimulation onset and the arrival of an antidromic action potential. In the case of somatic impalement, data for measuring several motor neuron electrical properties such as afterhyperpolarization were obtained as described previously for cat (Pinter et al. 1991).
To determine whether motor axon conduction velocities are slower proximally within the spinal cord or ventral root, we compared in several experiments conduction velocities measured from 20 MG motor neurons impaled within the spinal cord with conduction velocities determined by impaling 20 MG ventral root axons. Conduction velocities measured from motor neuron soma recordings are called "proximal" and include conduction through intraspinal and ventral root portions of the motor axon. Conduction velocities measured from ventral root axonal impalements are termed "distal" and do not include conduction through intraspinal and about half of the ventral root portions of the motor axon.
Single motor units were activated using 0.5-ms depolarizing current pulses delivered through the micropipette. Motor unit twitch data (twitch contraction force and time-to-peak) were collected first. Fused tetanic contractions were collected for stimulus trains of 50 to 200 Hz. Some HCSMA homozygote motor units fail to sustain fused contractions during tetanic stimulation, a phenomenon called tetanic failure (Pinter et al. 1995). Tetanic failure was measured as the difference between unit peak force and that present at the end of the stimulus train, normalized to the peak force and expressed as a percentage. Units showing a difference of 7% or greater are considered to exhibit tetanic failure. This percentage exceeds by 2% the estimated maximum variability of force baseline recordings. For units exhibiting failure, maximum tetanic force was measured as the peak force achieved during the axon or cell body stimulus train. Note that tetanic failure reflects the inability of a motor unit to maintain electrical activity of muscle fibers during an individual tetanic activation and does not reflect motor unit fatigue determined by muscle fiber fatigue as defined by Burke (1981).
Muscle samples obtained as described above were dissected further under a microscope into small strips in which nerve fiber bundles could be seen to pass among the muscle fibers. The strips were then stretched and pinned to the bottom of Sylgard-lined dishes. To prevent contractions, muscle fibers were crushed at sites approximately 2–3 mm on either side of the endplate region, which was reliably located in the middle of the muscle sample. The crushing produced muscle fibers with resting potentials in the range of −30 to −40 mV. Contractions generally were blocked for 2–3 h following the crush, after which new crushes were attempted or the muscle sample was replaced. These observations are similar to those made previously during EPC recording in crushed rat fibers (Argentieri et al. 1992). A concentric bipolar stimulating electrode (center pole diameter 250 μm) was used to stimulate the muscle nerve bundles with 0.1-ms duration constant current pulses. The output of the stimulator was capacitively coupled to the electrode to prevent passage of DC polarizing currents and to produce a bipolar-shaped stimulus current (Guyton and Hambrecht 1974).
Following pinning, muscle strips were stained with 10 μM 4-Di-2ASP to allow for visualization of endplates and muscle fibers (Magrassi et al. 1987; Rich and Lichtman 1989) and imaged with an upright epifluorescence microscope (Leica DMR). Once the endplate band was identified, a pair of glass micropipettes, filled with 3 M KCl, was used for separate voltage recording and current passage. The EPCs evoked by nerve stimulation were voltage-clamped using conventional voltage-clamp equipment (Axon Instruments, Foster City, CA). All EPC recording occurred within 250 μm of endplates, which provided full voltage control over the EPC amplitude as demonstrated by linear changes of EPC amplitude produced by varying the holding potential (data not shown). The electrode filling solution also contained 10 ng/ml sulforhodamine to enable fluorescence visualization of the electrode tips. Electrode resistances were generally about 10 MΩ. Holding potentials were usually −45 mV, but in several experiments −60 mV was used. In several instances multiple holding potentials were used to obtain current-voltage relationships, which were used to adjust the measured amplitudes of final records to different holding potentials. Final EPC records were averages of 16–30 sweeps collected at 0.5 Hz. To obtain records of potentiated EPCs, a nerve stimulus paradigm was used that was similar to the paradigm used previously to obtain records of potentiated motor unit twitch contractions in HCSMA homozygotes (Pinter et al. 1995). Trains of 20–30 nerve stimuli (150 Hz) were interleaved with single pulse stimuli at a rate of one stimulus event (single pulse or train) per 2 s until the EPC amplitude evoked by single pulse stimuli had reached a maximum. Under these in vitro conditions, this generally required four to eight stimulus trains.
In several experiments, current thresholds for action potential initiation were obtained from MG muscle fibers. Muscle fibers were not crushed for this recording, and dantrolene (6 mg/ml) was used to inhibit contractions. A pair of micropipette electrodes, filled as described above but with higher resistances (>20 MΩ), was used for separate voltage recording and current passage. In all cases, current injections were located within 250 μm of endplates. Current thresholds were determined using variable amplitude depolarizing waveforms that mimicked the shape of an EPC (time-to-peak, 1 ms; half-width, 3 ms; final time constant, 2.5 ms) and were applied to the current electrode by a D/A converter. In each examined fiber, threshold was determined by slowly increasing the amplitude of this waveform until an action potential occurred. Data were collected only from fibers with resting potentials more negative than about −70 mV.
All muscle fiber recording of EPCs and action potentials was performed at room temperature.
In one homozygous animal aged 144 days, MG motor neurons were intracellularly stained with the dye Neurobiotin. Following characterization of motor unit properties, depolarizing current pulses (400 ms duration, 5–10 nA, 1 Hz for 10–30 min) were used to iontophoretically inject 4% Neurobiotin (Vector Laboratories, Burlingame, CA) in 2.5 M KCl and 10 mM HEPES, pH 7.4. Following a 2-h delay from the final injection to allow for dye diffusion, the animal was transcardially perfused with PBS, followed by 4% paraformaldehyde in PBS. Motor neurons were injected with a minimum rostral–caudal spacing of about 200 μm.
Spinal cords were postfixed for 4 h and rinsed in PBS for 8–12 h, and vibratome sections (50–100 μm) were collected into PBS. Sections were permeabilized in 100% MeOH at 20°C, rinsed in PBS, and incubated in 50 μg/ml fluorescein-conjugated streptavidin (Molecular Probes, Eugene, OR) with shaking at 4°C overnight. After rinsing in PBS, sections were mounted on slides in an antifading medium (VectaShield, Vector Laboratories), and slides were stored at 4°C in the dark. Sections were imaged and analyzed using a confocal microscope (TCS-4D system, Leica, Eagle, PA) and interactive software. All confocal images illustrated are single-plane projections of a multiple-plane stack of images.
Following Neurobiotin staining and confocal microscopic analysis, coverslips were removed, sections rinsed in PBS, blocked in 2% BSA, 0.1% Triton X-100 in PBS, and immunostained using anti-phosphorylated neurofilament antibody (SMI34, 1:500 dilution, Sternberger Monoclonals, Baltimore, MD) followed by rhodamine-conjugated donkey anti-mouse IgG1 secondary antibody (Jackson Immunological). After rinsing, sections were recoverslipped in VectaShield, and sections were reimaged using confocal microscopy.
Following all imaging of fluorescence-labeled cells and processes, sections were processed for Neurobiotin visualization with peroxidase to enable detailed reconstruction under brightfield illumination and to avoid possible loss of structural detail because of photobleaching by prolonged fluorescent illumination. This was accomplished by removing the coverslips and processing the sections (on the slide) with an ABC kit (Vector Laboratories) and visualizing the peroxidase with 0.02 mg/ml diaminobenzidine (Sigma) and 0.01% H2O2 in 0.05 M Tris. The reaction was enhanced by adding 0.01% nickel ammonium sulfate (Alvarez et al. 1997).
Data from groups of animals were tested for phenotype-related differences (i.e., HCSMA homozygote versus normal) using nested ANOVA (Neter et al. 1990). Parallelism among linear regression slopes for individual experiments (i.e., individual animals) was tested using analysis of covariance. Phenotype-related differences in they-intercepts of linear regressions from individual experiments were tested using nested analysis of covariance. In some cases, differences between distributions were tested using the Kolomogorov–Smirnov test. All mean values are shown ± SE.
The results of previous studies suggest that motor unit dysfunction in HCSMA homozygotes is based on defective neuromuscular transmission (Pinter et al. 1995, 1997). To examine this possibility, we compared EPCs obtained from MG muscle fibers of 5 HCSMA homozygotes aged 118–144 days (average 132 days) with EPCs obtained from 5 genetically normal animals aged 146–167 days (average 152 days). All 5 of the examined homozygotes exhibited extensive weakness and atrophy of proximal musculature typical of HCSMA (Cork et al. 1982a; Pinter et al. 2001a). As shown in Fig.1, mean EPC amplitudes from homozygotes (−45 mV holding potential) were all less than the lowest mean EPC amplitude from control MG muscles. A nested ANOVA showed that mean EPC amplitudes from homozygotes were significantly less than normal (P < 0.01). We considered the possibility that this difference might be related to the age differences between the homozygote and normal populations. To test this, we performed a separate nested ANOVA on the group of data aged 143–148 days (which includes 2 homozygotes and 2 normal animals) and found that EPC amplitudes were significantly different (P < 0.01) despite the nearly identical ages.
Muscle fiber diameters measured during recording were on average smaller among homozygotes (30.4 μm ± 1.5, n = 5 homozygotes versus 38.2 μm ± 1.7, n = 5 normal;P = 0.01, nested ANOVA). Only weak correlations were observed between EPC amplitude and fiber diameter within experiments or within phenotype (homozygote, Pearson r = 0.09; normal,r = 0.16; P > 0.05). Thus differences in EPC amplitudes between homozygotes and normal animals are independent of differences in fiber sizes. Muscle fiber size differences between homozygotes and normal animals may be related to the age differences noted above or perhaps to the inability of HCSMA homozygotes to achieve the mobility of normal animals.
Associated with reduced EPC amplitude in homozygotes was the appearance of EPC failure in response to nerve stimulation at 2 Hz. While no EPC failures were encountered in 112 recordings from five normal animals, an average of 35% (range 14–62%) of EPC recordings in homozygotes (173 recordings) showed failure. Among individual EPC recordings that exhibited failure, failure occurred in about 15% of trials (±2.3%, range: 11–24%, n = 5 homozygotes). Overall, more than half (54%) of homozygote EPCs with mean amplitudes of 8 nA or less (−45 mV holding potential) exhibited failure in some stimulus trials.
An important question concerns whether homozygote EPC amplitudes are reduced in relation to threshold currents for action potential generation in muscle fibers. Such a relative reduction could help explain why motor unit forces of HCSMA homozygotes are reduced (Pinter et al. 1995). Since muscle fiber current threshold could not be obtained from crushed muscle fibers used to record EPCs, we compared the measured distributions of EPC amplitude with the distributions of current threshold obtained from uncrushed MG muscle fibers. In a subset of homozygous and normal animals, we measured MG muscle fiber action potential thresholds to current waveforms designed to simulate the shape of EPCs. A bar chart illustrating mean current thresholds sampled from ≥12 muscle fibers in two homozygotes and two normal animals is shown in Fig.2 A. A nested ANOVA indicated that no significant differences existed among these means that could be related to phenotype (homozygote versus normal, P > 0.05). No significant phenotype-related differences in resting membrane potential were found (P > 0.05), but there was a clear indication that differences in the resting potential present when the threshold was determined influenced the current threshold measurements (Fig. 2 B); the homozygote exhibiting the lowest average current threshold also possessed the most depolarized average resting potential. Further analysis of these data showed no significant differences in linear regression parameters relating threshold current (dependent variable) to resting potential among all examined animals (analysis of covariance for slopes and nested analysis of covariance for y-intercepts; both tests, P > 0.05). Data from homozygotes and normal animals were thus pooled to determine overall linear regression parameters. These parameters were used to adjust the measured values of current threshold to a uniform resting potential of −80 mV, which appears to be approximately the normal muscle fiber resting potential in dog muscle fibers as in other species (Rich et al. 1998). These procedures produced a mean value for MG muscle fiber current threshold of 50 nA and a SD of 16 nA.
To provide a standardized basis for comparing the derived current threshold distribution with measured EPC amplitudes, averaged linear regression parameters obtained from several samples of the relationship between EPC amplitude and holding potential were used to adjust measured EPC amplitudes to a holding potential of −80 mV. Figure3 A illustrates the distributions of these adjusted EPC amplitudes for five HCSMA homozygotes and five normal animals; data for each animal are shown in separate vertical lanes. To set a reasonable lower limit for the range of expected current thresholds at −80 mV, we selected a value of 2 SDs below the average current threshold (18 nA, see above). As shown in Fig. 3 A, almost all EPC amplitudes from normal animals are greater than this value, consistent with our observation that control animals exhibit no motor unit force failure (Pinter et al. 1995). In contrast, Fig. 3 A demonstrates that a substantial fraction of the EPC amplitude data from each homozygous animal is located below this threshold line.
We showed previously that both motor unit twitch force and EMG in HCSMA homozygotes can be dramatically increased immediately following trains of high-frequency stimulation (Pinter et al. 1995). The basis for this effect is likely due to a transient potentiation of EPC amplitude that enables activation of additional muscle fibers. To test this, we measured EPC amplitudes following application of the same stimulus paradigm used previously to demonstrate the presence of posttetanic potentiation of motor unit twitch force and EMG in HCSMA homozygotes (see methods). These data are plotted in Fig.3 B and demonstrate that, following potentiation, an increased fraction of EPC amplitudes obtained from homozygotes is above the threshold line. The percentage of EPC amplitude data above threshold in unpotentiated and potentiated states is summarized in Fig.3 C for homozygotes and normal animals.
These results indicate that, before potentiation, many motor terminals in HCSMA homozygotes are unlikely to produce sufficient current to enable MG muscle fiber action potential generation and that a large increase in the fraction of suprathreshold currents occurs following potentiation. Our analysis also indicates that muscle fiber current thresholds do not differ between HCSMA homozygotes and normal animals. It thus appears that decreased EPC amplitude is an important determinant of motor unit dysfunction in HCSMA. The observation that EPC failures in response to nerve stimulation are associated with EPC amplitude reductions in homozygotes suggests that motor unit dysfunction involves a presynaptic deficit of ACh release. An analysis of EPC quantal properties provided in the following paper supports this by showing that EPC quantal content at HCSMA homozygous motor terminals is significantly reduced relative to normal (Rich et al. 2002).
A previous study found that the size of ventral root motor axons obtained from forelimb spinal segments was reduced in HCSMA homozygotes relative to age-matched controls (Cork et al. 1989), and it was suggested that this might reflect growth arrest, axonal atrophy, or both. If decreased motor axon size occurs in association with the appearance of motor unit failure, then the reductions of synaptic transmission described above (which underlie motor unit failure) may reflect disruptions of axonal function rather than specific changes of synaptic function. The ideal approach for examining this possibility would be to study individual axon size in relation to the properties of single motor units. However, there are a variety of experimental difficulties associated with this approach, including the need for separate labeling of individual motor axons, since motor unit properties are studied in individual units functionally isolated by intracellular impalement of motor axons or cell bodies. As an alternative, we examined homozygote motor axon conduction velocity, since this is known to depend importantly on fiber size (Moore et al. 1978; Waxman 1980) and is readily measurable. To determine possible associations between changes of axon function and motor unit failure, we assessed when motor unit failure first appears in HCSMA MG motor units in relation to the time course of postnatal development of conduction velocity in MG motor axons.
Figure 4 A shows a plot of the fraction of sampled MG motor units that exhibit tetanic failure versus age for 25 homozygotes and symptomless animals. With one exception, the MG muscles of homozygotes older than about 120 days all possessed motor units that did not sustain force during repetitive activity. A considerable amount of interanimal variability in the fraction of failing MG motor units is evident in Fig. 4 A. Part of this variability is likely due to unavoidable sampling variations among animals. Part may also be due to varying expression of the disorder at the motor unit level for reasons that remain unidentified.
The time course of postnatal development of mean conduction velocity is shown in Fig. 4 B. Although an increased scatter of points may occur among older homozygotes (filled symbols, aged >120 days), average conduction velocity increased in a linear manner postnatally in HCSMA homozygotes. For homozygotes (60–188 days) and symptomless animals (60–140 days), mean conduction velocity and age were well correlated (Pearson r = 0.91, homozygotes;r = 0.92, symptomless; P < 0.01 both cases). Analysis of covariance showed that the slopes and intercepts of the fitted regression lines did not differ significantly (P > 0.05). The average values of about 85 m/s for both the oldest homozygotes (>180 days) and for two adult symptomless dogs from the HCSMA pedigree (aged >1 yr) agree well with reported values of MG motor axon conduction velocity in normal adult dogs (Lee and Bowen 1970).
To determine whether focal changes of conduction velocity might exist in proximal motor axons, we compared MG motor axon conduction velocities obtained from intracellular recordings made in the cell body with recordings made in ventral root motor axons in two homozygotes, both of which exhibited failing MG motor units (homozygote 1, 90% failing units; homozygote 2, 50%). In both cases, no differences in average conduction velocity obtained from each recording site were observed (Fig. 5; n = 20,P > 0.05, t-test). The greater mean values for homozygote 1 are most likely due its greater age (homozygote 1, 180 days; homozygote 2, 165 days). To confirm that these results are representative of normal, we studied two symptomless HCSMA animals of similar age and obtained an identical result (Fig. 5). These comparisons indicate that the conduction velocities of proximal motor axons (located between the soma and ventral root) do not differ significantly from the conduction velocities of more distal parts of the motor axons in HCSMA homozygotes that have failing MG motor units.
Overall, these data demonstrate that the development of axonal conduction velocity does not differ between HCSMA homozygotes and symptomless individuals and that the mechanisms producing reduction of synaptic currents and motor unit dysfunction in HCSMA homozygotes do not interfere with and act independently of mechanisms producing postnatal maturation of motor axonal conduction velocity.
In HCSMA and in other forms of motor neuron disease, swellings appear in proximal motor axons that are filled with misaligned neurofilaments and can achieve sizes as large as motor neuron soma (Cork et al. 1982b; Hirano 1991). To determine whether proximal motor axon swellings are associated with the occurrence of motor unit tetanic failure, we intracellularly labeled four MG motor neurons in one HCSMA homozygote (aged 144 days) after characterizing motor unit mechanical properties for each motor neuron. Three of four motor neurons were stained sufficiently well to enable visualization of the entire motor axon within the spinal cord, and all three of these cells innervated failing motor units. In two of the failing units, swellings were not visible in the labeled motor axons. Figure6 A illustrates EMG and force records from one of these motor units and demonstrates failure to sustain force output during repetitive activation (tetanic failure). Figure 6 B shows a low-magnification montage of several serial sections of the labeled soma, dendrites, and axon from the motor neuron innervating this unit and illustrates the absence of any swellings along the course of the axon from its origin to the ventral white matter. At higher magnification (Fig. 6 C), the axon could be seen originating from a primary dendrite and extending into the parenchyma without discontinuities or enlargements in its proximal portion. The functional properties of the motor unit and the structural properties of the motor neuron soma and axon from another failing unit were nearly identical (not illustrated).
The third labeled motor neuron exhibited small swellings in the proximal motor axon and showed the most extensive motor unit failure observed thus far in HCSMA. In this case, only the first two or three stimuli of a train of 60 depolarizing pulses (100 Hz) delivered to the motor neuron soma produced motor unit EMG potentials and almost no unit force was observed (Fig.7 A). The motor axon exhibited a smooth profile along its length except at the most proximal locations, where small swellings could be observed at low magnification (Fig. 7 B). At higher magnification, confocal imaging showed two distensions with outer diameters slightly larger than flanking axonal regions (Fig. 7 C, white arrows). These distensions are small compared with the sizes of axonal swellings that have been observed previously in the spinal cords of HCSMA homozygotes (Cork et al. 1982b, 1988), and we considered the possibility that this particular homozygote might lack large accumulations of neurofilaments in the spinal gray matter. As shown in Fig. 8, however, we verified the presence of abnormal accumulations of neurofilaments in adjacent sections of this spinal cord. These swellings immunostained positively for phosphorylated neurofilament (SMI31), and many were considerably larger than the apparent distensions observed in the failing motor neuron of Fig. 7. This suggests that, if the distensions seen in Fig.7 C are in fact accumulations of neurofilaments, they are most likely at an early stage in their development.
These data demonstrate that failure of motor unit function in HCSMA does not require the presence of neurofilament accumulations in proximal motor axons. These results also suggest that complete motor unit failure can precede the full development of neurofilament accumulations in proximal motor axons.
In normal animals, systematic patterns of correlations exist between and among motor neuron electrical properties and motor unit mechanical properties (Binder et al. 1996; Burke 1981; Pinter 1990). These relationships disappear after axotomy or after neuromuscular transmission is completely eliminated (Pinter et al. 1991; Vanden Noven and Pinter 1989). This functional organization forms an important part of the basis for the normal operation of motor neuron pools (Henning and Lomo 1985) and orderly recruitment of motor units (Cope and Pinter 1995; Zajac 1990). Others have suggested that these correlations may depend on trophic interactions between motor neuron and muscle that are likely mediated by mechanisms such as axonal transport (Mendell et al. 1994; Munson et al. 1997). We reasoned that, if any changes in the mechanisms underlying these correlations occur that provoke or are associated with the appearance of tetanic failure, then these correlations should be not be present or be diminished in data from HCSMA homozygotes. In the present study, we found significant correlations between MG axonal conduction velocity and motor unit mechanical properties in older homozygotes exhibiting tetanic failure as well as in other HCSMA animals exhibiting normal motor unit function. Table 1 summarizes average correlation coefficients for symptomless and homozygous groups for experiments in which five or more motor units were sampled and the fraction of these experiments in which significant (P< 0.05) correlations were observed between conduction velocity, twitch time-to-peak, and peak unit force measured during tetanic activation. An analysis of covariance that only included data from significant correlations showed that no significant differences in regression slopes or intercepts were present within either the homozygous or symptomless groups (P > 0.05).
Significant correlations were also observed between motor neuron soma afterhyperpolarization (AHP) duration and twitch time-to-peak in homozygotes (aged 80–177 days, r = 0.73,P < 0.01) and in young symptomless animals (aged 87–101 days, r = 0.79, P < 0.01). Analysis of covariance showed that the regression slopes did not differ between the homozygous and symptomless data groups (P> 0.05). These correlations and regressions were determined from data pooled from several experiments because AHP measurements were only made in smaller numbers of cells during individual experiments.
These data demonstrate that the relative incidence and extent of significant correlations between motor axon, motor neuron, and motor unit properties were quite high and independent of HCSMA phenotype. Since all but one older homozygote also exhibited tetanic failure among MG motor units, these data demonstrate that correlations between motor axon and motor unit properties persist despite the presence of motor unit dysfunction.
The results of this study provide several new insights into the pathogenesis of HCSMA at the cellular level. The first concerns the mechanism underlying weakness. Recordings of EPCs and muscle fiber threshold currents demonstrate that EPC amplitudes are reduced relative to EPCs from genetically normal control animals and to the currents needed to cause action potential firing in muscle fibers. A presynaptic deficit is implied by observations that the relative decrease of EPC amplitude in homozygotes is associated with the appearance of failures of EPCs in response to nerve stimulation. The immediate cause of motor unit dysfunction and weakness in HCSMA homozygotes is thus a reduced capability of motor terminals to release sufficient ACh to evoke muscle fiber electrical activity.
The demonstration that a large increase in the fraction of homozygote EPCs capable of evoking muscle fiber action potentials occurs following EPC potentiation helps explain two specific kinds of motor unit dysfunction observed in HCSMA. When first activated by intracellular motor neuron or motor axon stimulation, the twitch force and EMG of many motor units are often negligibly small but can be dramatically increased following tetanic activation of the motor neuron/axon or systemic administration of 4AP (Pinter et al. 1995,1997). These phenomena can now be understood to reflect the existence of EPCs at many motor terminals that are initially subthreshold for muscle fiber activation but become suprathreshold after transmitter release is increased by either tetanic stimulation or 4AP administration.
The second category of information provided by our results concerns the possible role of axonal abnormalities in the appearance of motor unit dysfunction in HCSMA. We examined how axonal conduction velocity matures in HCSMA homozygotes in relation to the appearance of motor unit failure. Motor axon conduction velocity is a functional attribute that depends on multiple factors, but among the most important are axon caliber and myelin thickness (Moore et al. 1978;Waxman 1980). Other studies indicate that cytoskeletal properties such as neurofilament number, density, and phosphorylation status are all important determinants of axon caliber and that myelinating cells influence these properties and perhaps neurofilament movement via slow axonal transport (deWaegh et al. 1992;Hsieh et al. 1994; Julien 1995;Williamson et al. 1996). Our results show that average MG motor axon conduction velocity in HCSMA homozygotes increases postnatally and reaches adult values by 180–190 days despite the earlier appearance of motor unit dysfunction (Fig. 4 B). The data also indicate that the rates of conduction velocity maturation do not differ between homozygotes and symptomless animals and that proximal and distal motor axon conduction velocities do not differ between these groups. These results show that axonal conduction velocity continues to develop normally in HCSMA homozygotes despite the appearance of motor unit failure. Because of the dependency of conduction velocity on multiple factors, complicated interpretations of these results are possible. However, the simplest view is that the large variety of mechanisms responsible for axonal maturation remains unaffected by and is independent of the mechanisms that give rise to reduced EPC amplitudes and motor unit dysfunction in HCSMA homozygotes.
We also considered the possibility that abnormal accumulations of neurofilaments or other focal disruptions of function in proximal motor axons might be associated with or cause motor unit failure. Our intracellular labeling experiment enabled us to examine directly for the first time the proximal axons of diseased motor neurons with known motor unit dysfunction (Figs. 6 and 7). Although the data are from one homozygote, they constitute an existence proof that neither the onset of motor unit dysfunction nor its evolution in HCSMA requires swellings in proximal axons. In the case in which evidence of proximal axonal swellings was observed (Fig. 7), the extent of motor unit failure was nearly complete but the swellings were small compared with the size of accumulations that could be demonstrated in adjacent spinal cord areas (Fig. 8). These results suggest that neurofilament dysfunction as manifested by aggregations in proximal motor axons may not occur until after the motor unit has lost its ability to produce useful force output. Overall, our data suggest that failure in cellular systems that support maturation of the motor axon and mechanisms that produce neurofilament swellings in motor axons do not give rise to reduced EPCs or motor unit tetanic failure in HCSMA. This is consistent with our recent finding that tetanic failure in HCSMA occurs in the absence of evidence for degeneration at neuromuscular junctions (Balice-Gordon et al. 2000).
Our finding that MG axonal conduction velocity matures normally in homozygotes despite the appearance of motor unit failure is in apparent conflict with the study of Cork et al. (1989) who reported that motor axons in HCSMA homozygotes fail to achieve normal size during postnatal development and that some may exhibit atrophy. HCSMA features a stereotyped, spatial–temporal pattern of progression in which the innervation of axial and proximal muscles shows the earliest onset of involvement and is always more progressed than in distally located muscles such as the MG (Balice-Gordon et al. 2000; Pinter et al. 2001a). One factor that may thus have contributed importantly to the results of Cork et al. is that axon size was studied in ventral roots of forelimb spinal segments. These roots most likely included a mixture that included axons supplying neck and rostral paraspinous muscles that are extensively involved at the ages studied. In contrast, the present study focused on a single, distally located muscle that becomes involved later in the course of the disease. Another contributing factor appears to be the presence of an exceptionally large value of axon size for one control animal that probably exaggerated differences between homozygote and normal animals (see Table 1 of Cork et al. 1989). Axonal atrophy and degeneration are components of HCSMA, and it is thus likely that conduction velocities eventually decrease. The present results indicate, however, that the time course of the pathogenesis is such that these events occur after motor unit function is lost because of neurotransmission reductions and that the mechanisms mediating axonal atrophy and degeneration do not play a key role in determining loss of motor unit function.
Correlations between motor neuron and motor unit properties
Significant correlations between MG motor axon and motor neuron properties and motor unit mechanical properties were found in HCSMA homozygotes and were similar to those found in symptomless animals (Table 1). Previous studies have shown that these correlations appear within several weeks after birth in the cat (Bagust et al. 1974) and the same seems likely for the dog. Our data thus demonstrate that motor unit dysfunction occurs in HCSMA homozygotes without disturbing these relationships. Correlations between motor neuron and motor unit properties represent an important part of the functional organization underlying orderly recruitment of motor units (Binder et al. 1996; Burke 1981;Cope and Pinter 1995; Pinter 1990). The presence of these relationships in older HCSMA homozygotes contributes to the sense that MG motor unit recruitment order and usage pattern are likely to be normal, at least at the time when failure begins to appear. The mechanisms that establish and maintain correlations between motor neuron and motor unit properties are not known. Some have suggested that trophic interactions between motor neurons and muscle are responsible at least in part (Mendell et al. 1994;Munson et al. 1997). Following axotomy, correlations among motor neuron properties disappear (Pinter and Vanden Noven 1989). Because these correlations are present among the motor units we sampled, the onset of tetanic failure does not appear to be associated with an axotomy-like response among MG motor neurons in HCSMA. The postnatal increase of average axonal conduction velocity (Fig. 4) also supports this point. By contrast, axonal injury leads to a reduction of conduction velocity (Pinter and Vanden Noven 1989) that is associated with a decrease in axon caliber and in neurofilament protein synthesis (Hoffman et al. 1988). One factor that may inhibit the appearance of axotomy-like changes in motor neurons innervating failing motor units is the persistence of neurotransmission to a few muscle fibers, since axotomy-like changes do not appear in normal motor neurons even when most synaptic transmission with muscle is blocked (Pinter et al. 1991).
Our work in HCSMA demonstrates that diminished neuromuscular synaptic transmission provokes significant or complete loss of motor unit function well before the appearance of axonal neurofilament swellings or evidence of axonal dysfunction, including motor terminal degeneration (Balice-Gordon et al. 2000). Since neurotransmission failure appears to be independent of axonal maturation, it is conceivable that the underlying defect may be specific for mechanisms located in the nerve terminal. It is not yet known whether neurotransmission failure underlies loss of motor unit function in other forms of motor neuron disease, although there is evidence in amyotrophic lateral sclerosis that neuromuscular synaptic dysfunction occurs while motor axons can still conduct action potentials (Maselli et al. 1993). It has also been reported that degeneration of motor terminals occurs well before motor neuron cell death in the SOD1 transgenic model of familial amyotrophic lateral sclerosis (Frey et al. 2000). These observations raise the possibility that inhibition of motor neuron cell death and perhaps even axonal degeneration may not be sufficient to prevent loss of function in motor neuron disease. Indeed, experimental treatments of motor neuron disease that inhibit cell death and axonal degeneration but fail to provide significant and lasting functional benefit might be explained by the persistence of additional but untreated mechanisms that compromise neuromuscular function or structure and thus motor unit function (Klivenyi et al. 1999). This view is consistent with emerging evidence that survival or maintenance of the motor terminal may be regulated differently and independently of mechanisms controlling survival of the axon and cell body (Gillingwater and Ribchester 2001). Further insights into the mechanisms underlying loss of neuromuscular function in motor neuron disease may be necessary to accomplish effective treatment. The following paper provides additional insight into possible mechanisms underlying loss of neuromuscular transmission in HCSMA (Rich et al. 2002).
We thank D. Dewey, D. Smith, and A. Shirley for technical assistance.
This work was supported by National Institute of Neurological Disorders and Stroke Grants NS-31621, NS-07287, NS-31563, and NS-25547.
Present address of R. F. Waldeck: Department of Biology, University of Scranton, Scranton, PA 18510.
Address for reprint requests: M. J. Pinter, Department of Physiology, Emory University School of Medicine, Whitehead Bldg., 615 Michael St., Atlanta, GA 30322 (E-mail:mpinter{at}physiol.emory.edu).
- Copyright © 2002 The American Physiological Society


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Falling for Christmas(2010)
Cedar Cove Christmas / Call Me Mrs. Claus
An omnibus of novels by Debbie Macomber
Packed with festive romance and fun, these two seasonal stories will melt any heart this Christmas. "A Cedar Cover Christmas" - Mother-to-be Mary Jo Wyse arrives in Cedar Cove on Christmas Eve, searching for her baby's father. David Rhodes had said he'd be in town. But he isn't, which leaves Mary Jo stranded, pregnant and alone. And there's no room at the local inn...However, the people in Cedar Cove are all willing to lend a hand ensuring there will be plenty of faith, hope and love this Christmas. "Call me Mrs. Claus" - Clare Keneally is a long-divorced maths professor with four adorable - and very mischievous - grandchildren. With Clare looking after the children for the holidays, their grandfather Jim also drops by and this starts the kids thinking...wouldn't Christmas just be perfect if they 'helped' their grandparents fall in love and get married again?
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September 2010 : Canada Paperback
|Title: Falling for Christmas
Author(s): Debbie Macomber
ISBN: 0-7783-0394-2 / 978-0-7783-0394-7 (Canada edition)
Publisher: Mira Books
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Bertrand Burgalat Cabaret du Mile End, Montreal, QC, September 21
Published Sep 22, 2012
Burgalat played his first-ever show in Montreal to an enthusiastic crowd that eagerly welcomed the singer's old repertoire as much as recent numbers like "Bardot's Dance" and "Survet' vert et mauve." Cabaret du Mile End was indeed a pretty good place to forget about the Mayan calendar last night. The 49-year-old singer, producer and composer, backed up by four musicians, played an eclectic mix of keyboard-driven pop songs that owed as much to Jean-Claude Vannier as to Claude François.
Best of luck, Mr. Burgalat, maybe someday critics will stop focussing on the ascot and glasses to pay a little more attention to details that don't call for the winning word "dandy" tossed at every occasion.
To see Exclaim!'s Pop Montreal photo gallery, head here.


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Grammy Award-winning singer Mariah Carey made her concert a family affair in Queensland, Australia.
After husband Nick Cannon warmed up the crowd with a DJ set, the couple's 1 1/2-year-old twins Moroccan and Monroe took to the stage. "Ms. Monroe greets the audience in Australia," the proud mom tweeted. "Thank u 4 coming!!"
On Saturday we spotted the Carey-Cannons sailing around the Great Barrier Reef.
The American Idol star and the America's Got Talent judge have just celebrated "the most festive Christmas yet" in Aspen, Colorado with their twins, "Roc and Roe."
The votes came pouring in for "Dem Babies" in our 5th Annual Readers' Choice Awards! They were our favorite toddlers of 2012!


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Snow has been falling hard in Steamboat. "Steamboat has some of the best snow in the Rocky Mountains and is currently enjoying mid-winter conditions even though the calendar says spring," said Chris Diamond, president and chief operating officer for Steamboat Ski & Resort Corporation.
Steamboat has consistently received snow all winter. The mountain tallied snow on 70 out of 125 days with 31 of those days recording four or more inches. Locals love it!
"Spring is the best time of year to gather friends and family and spend time on the slopes," said Diamond. "This is the time of year when everything lines up just right: great snow, longer days, incredible views, and plenty of fun."
It's also Steamboat's 50th Anniversary Season. Live music will resound from the base area with the Bud Light Rocks The Boat FREE Concert series.
But the big celebration is Billy Kidd's 70th birthday. Billy Kidd is Steamboat's Director of Skiing. He's also a living legend as he is an Olympic Silver medalist and World Champion. His birthday is April 13, which is the day of the Cardboard Classic. The next day is closing day and also when the mountain hosts the 6th annual Splashdown Pond Skim at the base area.
Another crazy event that has quickly become a Steamboat tradition is the Poochy Paddle at the Old Town Hot Springs. It is a bi-annual event which is 2-4 p.m. Sunday, April 21. Dogs are invited to take a dip in the lap pool of the Old Town Hot Springs, before it is drained and cleaned for the spring. A couple hundred soggy doggies love the fun, which includes plenty of tennis balls and Frisbees. The cost is $5 per dog before the event and $10 per dog on the day of the event.
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You are here: Home → Forum Home → Bumblebee forums → Bumblebee discussion → Thread
Has anyone seen any Bombus lapidarius , red-tailed bumble workers or Queens??
I live in Swindon and i haven't seen one yet. I know we've had awful weather this year but wondered if they could still appear even though it is later than usual.
Please say if you have seen any in your area, thanks all.


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-That recent Washington Post poll which showed Mayor Adrian Fenty trailing by double figures in his race against challenger Vincent Gray?
That wasn't the only surprise in the poll.
Try the race for the Democratic Party At Large Council seat currently held by 12-year-veteran Phil Mendelson.
For as long as his challenger—relative newcomer Clark Ray—has been running, which is approximately a year, you'd have thought that this was a race between Mendelson and Ray, experience vs youth, twelve years of service vs. an impressive resume.
Guess again.
You'll never guess who was leading the race according to the Washington Post poll. Guy by the name of Michael Brown. Better yet, call him Michael D. Brown, by which middle initial the current Democratic Shadow Senator can be differentiated from the other Michael Brown, (Michael A Brown) the at large Independent member of the council member who is not running this time around.
Mr. Brown, who didn't enter the race until late this summer, raised no money and had no Web site, led Mendelson by 38-21 percent among all voters and 41-29 percent among likely voters.
Ray, who had been running hard throughout the campaign, was mired in single figures.
Brown, Michael D, in fact, registered high among African American voters, where he got 49% percent of the
vote. His name on the ballot says simply Michael Brown.
The news almost made hash of the at large campaign. "Well, sure, it's had a big effect," Mendelson said. "But we're moving to correct it on every possible front."
At a recent candidate forum (which also included Ward 1 and the Chairman's Race) in Adams Morgan, Brown, Michael D, professed not to be surprised by the events.
"I think people like my story, like my message," Brown, who is a successful businessman, after having dropped out of high school, which he had called the biggest mistake of his life. A big, affable man who's lobbied hard for statehood, and jobs for DC workers, as well as the district having a voting power and taxing power over suburban workers in DC, said. "There's no subterfuge. I've used that name all of my life. It's my name. I don't really think people are confused."
A local resident disagreed. "I think what you're doing is misleading and dishonest," he said to Brown. "It's not fair to the other candidates or voters."
Mendelson, who clearly thought he'd be ahead in this poll, moved quickly to staunch the crisis. "I've been representing voters for 12 years now," he said. "People know they can rely on me. But this, well, I just think this was something that was clearly misleading, clearly a mistake. He was at one forum where Brown was pointed out to a man who had voted for him in a straw poll and the man said he was voting "for the other Brown." Michael A Brown has sent out recorded calls to voters saying he's not running
Mendelson has passed out flyers showing pictures of himself, of Michael D. Brown and Michael A. Brown. For one thing, Michael A. Brown, who ran unsuccessfully for mayor, and who is the son of the late U.S. Commerce Secretary Ron Brown, is black which Michael D. Brown clearly is not.
"You have to get the information out there," Mendelson said in an interview. "It's a problem. No question. But it's obvious that the poll shows two things: voters haven't been paying enough attention because the mayor's race has taken up all the focus of voters, and they obviously confused this Brown for Councilman Brown, who is not running."
"I thought we were running against Clark Ray," he said. "But I also think it's obviously hurt him (Ray) more than it hurt me."
Mendelson has often opposed Mayor Fenty, especially on school reform. He voted against the mayoral takeover. Surprisingly though, he has some different views about what to do with Chancellor Rhee. "I wouldn't propose firing her," he said. "I'd want to keep her here. I'm for reform, but didn't agree with how she did things." But I think we ought to stick with continuity. We can't keep doing this, firing people after a year or two and starting all over again. that's not good for the kids."
Ray, who looks boyish with a bald look, is sometimes confused not with any Browns but with the mayor. He said he was "disappointed" in the results. "Look, I'd be lying if I said I wasn't both surprised and disappointed," he said. "But I can sleep at night, pretty easily."
Ray has run with energy, intelligence, and great enthusiasm. He brings a big resume to the table—services with the Clinton Administration in the White House and the Department of Agriculture, a former teacher, and a former Head of the Parks and Recreation Department under Fenty, whom he still admires, even though Fenty fired him. "I don't want to get into that, in terms of feeling, it's something that's between him and me," he said without a trace of bitterness.
Of the polls, he said that there's still a large bundle of undecideds to account for. "I'm not discouraged," he said. "I'm going to keep on doing what I'm doing and I'm going to run as hard as I can up to the last moment.
At the forum, the candidates were asked about what their biggest mistake was and how they handled it. Brown, D, said it was dropping out of high school, and then talked mostly about overcoming that mistake. Ray remembered eagerly proposing a project in his role as parks and recreation director that would have eliminated some 20 jobs. "I didn't know that at the time," he said. "The lesson is that there are consequences to policy decisions. That's a hard lesson, but it's not something you forget."
The rise of Brown, which in some ways almost undermined the, months of campaigning that had been done by Mendelson and Ray, didn't faze him.
"I kind of think of it as poetic justice," he said. "I mean, here's Phil, who depends hugely on name recognition for twelve years, and here he's become a victim of it.
The subject never came up during the forum. Well, almost. "I realize that we have a problem over some confusion about names," Ray said to the forum audience. "Well, I want to make it absolutely clear . . ., it's Clark Ray. …Not Ray Clark. People make that mistake all of the time. So I wanted to clear that up."


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text:
Human eyesight can fall prey to all manner of optical illusions, but in this case the object really is as huge as it appears.
Racing into a corner with Boston Bruins' defenceman Zdeno Chara, the tallest man in the NHL, is not for the easily ruffled, regularly doing so when you're five-foot-seven (officially, anyway) requires nerve, savvy and a dose of reckless disregard.
"Ah, it's second nature, I've been doing it for a long time," said the Montreal Canadiens' Brian Gionta. "There's no denying how strong he is, and how big, you just have to use your assets."
To do battle with Chara, seven feet tall on his skates, is to match wits with a guy with beast-of-burden strength who has a freakishly long reach; he is to be approached using speed, and quickness.
"The problem is when he gets hold of you, he can overpower you. It's a delicate balance, how you play against him, where you find your seams," Gionta said.
While Chara undoubtedly has that long stick, and isn't shy about using it to shove, lean on, and otherwise abuse adversaries, he is about as nimble as you'd expect that large a man to be.
So players like Gionta, Brendan Gallagher, David Desharnais, and Daniel Brière – all members of the Habs' 5-foot-10, 180 pounds and under brigade – typically like to play in close proximity to the big man, inside his reach.
It would be churlish to describe it as the worm's eye view (is it nicer to call it the Chihuahua's?), the sense from the Habs' smaller forwards is that while Chara is not an easy guy to face, they don't hate the challenge.
"You get a pretty good idea [of his size] from watching … but everyone has strengths and weaknesses and you just try and play to your strengths," Gallagher said. "Regardless of who the defenceman is, I'm going to be the smallest guy going into the battle. It's something I've dealt with my whole life, you understand how."
In the 5-foot-9 Gallagher's case, the how involves quick pivots, tenacity and a facing-the-boards game that is surprisingly strong for a smaller player. Chara, who generally plays half the game, is the Bruins' defensive linchpin.
However, Chara is not the only player who poses a defensive obstacle to the Habs. Skating through the stingiest defensive team in the Eastern Conference only gets you as far as goalie Tuukka Rask, a Vezina trophy finalist.
But to suggest, as conventional wisdom holds, that this is a battle of a big, brawny team and a small, fleet one ignores the facts.
On average, the Bruins are taller than the Habs, it's true, but only by 1.4 inches(Boston's average is surely skewed by Chara and six-foot-five Dougie Hamilton); they're also heavier, by one whole pound.
With players like Max Pacioretty, Rene Bourque, Lars Eller and noted Bruins-killer Thomas Vanek (30 goals, 62 points in 55 career games against Boston) in their top nine forwards – and a speedy fourth line that exceeds the Bruins' skill-wise – Montreal isn't as easy to push around as many might think.
The stereotype of Boston as a massive, slow-moving squad is also unfair – Habs coach Michel Therrien said "they don't just rely on size, they've got speed too."
While Boston will be favoured by the odds-makers, the two teams have posted nearly identical records since March 15 (Montreal has actually won one more game over that stretch).
Both squads are deep, if there's an obvious point of differentiation, it may be that the Habs, by their mere presence, drive the Bruins' blood pressure upward.
Forward Brad Marchand described Montreal this week as "one of those teams you just want to hit."
Boston coach Claude Julien used the word "hate" in the same sentence as the Canadiens on Tuesday – although in fairness, he said he also used to hate the Bruins when he was paid to coach the Habs – and it's an emotion that seems to be more keenly felt in the north end of Beantown than it is in Montreal (even if Therrien said "no comment" when asked about the hatred level he has for his rivals).
When it was suggested to Habs goalie Carey Price the Bruins have a white-hot loathing of their opponents, he deadpanned "that's not very nice."
"I'm sure they're going to be a focused group. That team is filled with veterans and they know how to win in the playoffs," said Price, who is on friendly terms with Rask, his fellow 2005 first-rounder. "We can't fall into any kind of, I don't know how to put it, circustry? You can coin that one."
But if the Habs' singular ability lies in coaxing the Bruins' players to drive to the outskirts of Sanityville, that's fine too.
"Whatever it takes, I suppose. We have a bunch of guys in this locker room that are very tenacious and tough to play against. That tends to drive other teams crazy, and there's no sense in changing what we do well," Price said.
There is, of course, a great deal of history between these two teams in the playoffs, and perhaps it's true the Habs have some mysterious mojo advantage, having won 24 of the 33 postseason meetings between the teams.
The Bruins have won the last two, and will be looking for cosmic redress; Montreal and Boston always seem fated to collide in the big moments.
It's as it should be, really.Report Typo/Error
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Checked the team store and they didn't have it? anyone know?
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Checked the team store and they didn't have it? anyone know?
Contact the Home Court Gift Shop by phone (the one at Conseco Fieldhouse). I know (well, I'm 99% sure) they ship items out. Ask them how much it would cost to ship to where you live, and if the price is right I'm sure they'll have yellow jerseys to choose from; they were loaded with them when I was there Sunday.
Otherwise ask if someone here is willing to ship it to you cheap if you give them the funds to pay for the jersey themselves first.
There are probably quite a few at the Salvation Army.
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The ACT is the only jurisdiction to go backwards on a national scorecard for residential building activity.
Queensland and Tasmania sit below the ACT on the Housing Industry Association's half-yearly league tables, which are based on 14 key indicators.
But those states are little changed when compared with long-term averages, whereas the ACT tumbled from fourth to sixth place on the table and could still be showing after-effects of the boom years of 2010 and 2011 for new home building.
On the other hand, the NSW home building market appears to be gathering momentum after years of under-building, and is attracting buyers from the territory, according to the HIA.
Association economist Geordan Murray says the ACT's scorecard is the worst it has been since 2008, although in such a small jurisdiction trends could be volatile.
"Lower levels of activity in the multi-unit segment drove the decline, although there were also emerging signs of weakness in the ACT's detached-house building market," Mr Murray said.
"On the chart showing where changes have occurred, the value of alterations and additions declined two ranking points, approvals for detached dwellings dropped three points, and another big mover down is turnover of established homes, which dropped three points," he said.
"We look at the number of transactions for established homes and transactions involving multi-units, and the number of hours worked for workforce. The ACT has performed poorly."
One bright spot was the ACT's detached-house segment, for which it was the top-ranked jurisdiction. But a lower ranking for detached-dwelling approvals suggests this position may not be maintained.
Cutbacks in the public service and a decline in real wages for those still employed in the public service had caused the slide.
"The uncertainty within the household sector is clearly evident in the figures related to turnover of established homes, " Mr Murray said.
"A decline in turnover saw the ACT's ranking for established-house transfers slip three places while the ranking for transfers of established multi-unit dwellings slipped one place."
Association ACT and southern NSW executive director Neil Evans said people were going across the border to build new homes.
"You only have to look at Googong and Tralee, it's a real option. We have a situation where we are surrounded by NSW, where the housing market is really starting to bounce back because they have been under-building for nearly a decade and the NSW Government has good incentives for first-home owners and land is much cheaper," he said.
"It is not dire straits [in Canberra] by any means," Mr Evans said. "We are still averaging 3200 [dwellings] a year and our 10-year average is 2800.
"We are still doing OK but we did come off an almighty high from 2011 when we were doing 5000 dwellings."
Mr Evans said that after coming back from that spike, he would like to see the market perform a little better, and the HIA was looking at policies to reinvigorate activity in the ACT.
Mr Evans said after the boom years of 2010 and 2011, he was sure some contractors, sub-contractors and their employees would be moving interstate for opportunities from NSW's momentum.
NSW's ranking on the HIA scorecard sits at third place behind Western Australia and the Northern Territory. It has experienced strong multi-unit activity, which is beginning to lose momentum.
Mr Evans said the ACT's renovations remained reasonable.
"We are looking at doing around $380 million of renovations a year, that is what is recorded by the [Australian] Bureau of Statistics, there would probably be that figure again in small renovations and repairs and maintenance that doesn't need a building approval."


text: Blogs Joe Weiss Testifies! Category: 2009 Acs Conference ACS Joe Weiss Senate Testimony Submitted by waltboyes on Sat, 03/21/2009 - 13:27 We've managed to extract from the Senate's streaming video feed only that part of the hearing that consists of Joe's testimony. The previous post gives the information on where to see the entire hearing, but we thought we'd give Joe's testimony itself some blog time. Login or register to post comments Read more 3380 reads Permalink


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It is hard not to be cynical about the recent announcement by Walmart, the world's largest retail company, that it plans to use its enormous "size and scale to help empower women across its supply chain".
The scheme – which Walmart has dubbed its Global Women's Economic Empowerment Initiative – involves doubling the money the company spends with women-owned businesses, providing women around the world with job training and access to education and approaching its biggest suppliers to use more women and minorities on work they do for the company.
These are noble goals, and I am fully in favour of policies that support women-owned businesses and programmes and improve the lives of working women. But coming just a few months after the US Supreme Court threw out a class-action sex discrimination lawsuit against Walmart, the initiative smacks of a PR stunt.
The lawsuit alleged unfair treatment of women in the workplace and cited a pattern of discrimination in pay and gender-biased promotions at Walmart. Writing in the FT in June, Philip Delves Broughton wrote: "women at Walmart fill 70 per cent of the hourly jobs in its shops, and make up only 33 per cent of management. Women are paid less than men in every region and the salary gap between men and women widens over time."
However, the court ruled that Walmart's women workers did not constitute a single class for the purposes of a discrimination suit.
Ted Boutrous, Walmart's lawyer, said at the time, the ruling would be "very helpful in bringing back sanity to class action law in all different types of areas."
Fast forward to September and the company is championing a new plan to make sure that "women view Walmart as a retailer that is relevant to them and cares about them," according to Mike Duke, the company's chief executive.
I spoke to Detlev Suderow, a senior lecturer at Brandeis International Business School, and an expert in the field of human capital management, to get his reaction to Walmart's news. He told me that the programme is an "underwhelming PR action".
"The job of the HR department is to make sure that its company has fair and equitable hiring and advancement policies, not only because it is the right thing to do, but because it keeps their companies out of legal and financial trouble."
One component of the programme is a pledge to "empower women through job training and education". The company plans to help 200,000 women from low-income households around the world to gain job skills and access higher education.
"They are spending a lot of money training women in factory and retail jobs around the world, but traditional labour is changing and we are going to have many fewer women who are willing to take that kind of job," says Suderow.
"Companies will soon be facing a shortage of human capital, and anything Walmart can do now to build relationships with low-cost human capital is a smart move, but this is nothing breakthrough. This is just what strategic HR is supposed to do."


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HOUSTON - A small northeast Houston grocery store had to shutdown after an early morning fire Tuesday.
Houston firefighters were called to the Queen Saver Supermarket on Huffman and Cavalcade around 4:30 a.m.
It took crews about 15 minutes to get the flames under control.
Upon closer inspection, there were signs the fire may have started from an outside electrical box.
Investigators believe it spread up an outside wall and into the attic.
The store was so badly damaged, it was closed for repairs.
Firefighters said it could take days to get the store repaired and ready for an inspection.
The store was closed when the fire started, so no one was injured.
Copyright 2013 by Click2Houston.com. All rights reserved. This material may not be published, broadcast, rewritten or redistributed.


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Most orders ship within 1 business day. Shipping times for:
* Excludes Alaska and Hawaii
Even big or heavy items, like rollators and wheelchairs, ship free. If your order
total is $75+, the shipping's on us!
Extra Wide Tall-Ette Elevated Toilet Seat (with or without Legs)
Stander EZ Adjustable Bed Rail with Padded Pouch
Drive Exercise Peddler
Omron 10 Series Upper Arm Blood Pressure Monitor
Rodger Wireless Bedwetting Alarm System
Medihoney Manuka Hydrocolloid Wound Fill Paste - 1.5oz Tube
Drive Adjustable Seat Height Rollator
Omron Micro-Air Electronic Nebulizer System NE-U22V1
Stander BedCane and Organizer Pouch
Regular Price:
Special Price:
The Exercise Peddler is an excellent rehabilitation and exercise device. It is completely ideal for toning leg and arm muscles. The Exercise Peddler allows a safe and gentle form of low-impact exercise with adjustable tension to suit individual preference. Due to its small size and light weight, it can be easily moved and adjusted to permit the user to operate and store it with ease.
It comes completely assembled with four anti-slip rubber pads to prevent sliding and to protect floors. On top of its immediate practicality, it also features an attractive silver vein finish and comes with a limited lifetime warranty, making it a truly smart buy.
Question will be displayed after moderation.
Got this to exercise while unable to go outside. The set up was simple and it is a good way to get some excercise while sitting. The operation is smooth and it is heavy enough to stay in place.


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Diet Lemonade
Cast away the calories from sugar, and sip your way through summer with diet lemonade. From drink versions made with fresh lemon juice to other drinks spiked with cayenne, these diet lemonade recipes are sure to expand your love of lemons.
Tangy Citrus LemonadeFitness
Limes add a little tartness to this summer drink. Fresh raspberries add pretty color.


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FDA phasing out asthma inhalers using CFC propellant
UPDATE 7/22/10: Please note that the LA Times ran this story with incorrect dates (as seen below). The correct dates as given by the FDA are:
The affected products and their phase out schedule include:
|Inhaler Medication
|Last Date to be manufactured, sold or dispensed in U.S.
|Tilade Inhaler (nedocromil)
|June 14, 2010
|King Pharmaceuticals
|Alupent Inhalation Aerosol (metaproterenol)
|June 14, 2010
|Boehringer Ingelheim Pharmaceuticals
|Azmacort Inhalation Aerosol (triamcinolone)
|Dec. 31, 2010
|Abbott Laboratories
|Intal Inhaler (cromolyn)
|Dec. 31, 2010
|King Pharmaceuticals
|Aerobid Inhaler System (flunisolide)
|June 30, 2011
|Forest Laboratories
|Combivent Inhalation Aerosol (albuterol and ipratropium in combination)
|Dec. 31, 2013
|Boehringer Ingelheim Pharmaceuticals
|Maxair Autohaler (pirbuterol)
|Dec. 31, 2013
|Graceway Pharmaceuticals
From Los Angeles Times:
The Food and Drug Administration said Tuesday it is taking a long-expected step and phasing out the production and sale of asthma inhalers using chlorofluorocarbons as a propellant. The chlorofluorocarbons, commonly known as CFCs, were once widely used in a variety of applications, especially as refrigerants, because of their inertness, but they have been shown to damage the Earth's ozone layer, which protects life from the damaging effects of the sun's ultraviolet rays. Most uses of the chemicals have already been abandoned. Medical devices employing them are among the last to be affected.
Four of the seven devices using CFCs are no longer being made, but they are being banned to prevent their reintroduction. The rest will be forbidden after the end of 2013. Patients using inhalers will be able to buy alternative inhalers containing the same drugs but using different propellents – most notably hydrofluoroalkane, which is destroyed in the atmosphere before it can reach the ozone layer.
The devices that are no longer being made and whose sale will be forbidden after June 14 are:
- Tilade Inhaler, made by King Pharmaceuticals
- Alupent Inhalation Aerosol, made by Boehringer Ingelheim Pharmaceuticals
- Azmacort Inhalation Aerosol, made by Abbott Laboratories
- Intal Inhaler, made by King Pharmaceuticals
The three products whose sale will be permitted until Dec. 31, 2013, are:
- Aerobid Inhaler System, made by Forest Laboratories
- Combivent Inhalation Aerosol, made by Boehringer Ingelheim Pharmaceuticals
- Maxair Autohaler, made by Graceway Pharmaceuticals
The agency cautioned against buying any of the banned products over the Internet because they are often mislabeled or do not contain effective ingredients.
Original here.
View the FDA news release.


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mixed bag: Celebrity Infinity Cruise Review by shaakey
Overall Member Rating
mixed bag
Destination: Alaska
Embarkation: Seattle
We arrived at the port at about 11 a.m. We were able to board immediately. We were told to go to the buffet until our rooms were ready. Upon entering the buffet, we saw a very large selection of food served in a smorgasbord fashion. Although there was a large selection and it seemed that the charm was more in the unlimited amount, rather that the quality of food. Still...we were going to Alaska and were filled with excitement as we ate.
An announcement was made that the rooms were ready so we proceeded to ours. Upon entering our room, my girlfriend and I noticed a pretty strong odor of stale cigarette smoke. I am an asthma sufferer and found myself starting to wheeze. I went to More guest services and explained our dilemma. I was told that there no rooms designated as "Non Smoking" on the ship, and they would wipe down the room as to eliminate the odor. Once we were underway, they would look for another room. As I waited my asthma increased and this affected my whole trip.
Our ship started moving, our phone rang. It was guest services letting me know that they found us another room, just a few doors down. I said we would re-pack to move. She said there would be no need as she would send someone down to move us as we were. A couple minutes later there was a knock at the door. We opened it to find our room steward, who looked into our cabin and told us that we needed to pack so we could switch rooms. We told him that guest services had advised us that we would be moved without having to re-pack. He explained that he could not let us into the new room until everything was out of the old room. I was not comfortable leaving our belongings in the hall, so I called guest service back. Again they told me not to not to lift a finger, as they would have someone come right away to move us. Another knock on the door and our room steward appeared again, only this time he seemed a bit miffed. We managed to get into our new room, and as we settled in we again felt the excitement that brought us here in the first place.
Our first port Sitka was breathtaking!! Once we got off the ship, we proceeded to go past all the shopping and walked up the main street until we came to a lake. We sat there on a park bench and had a great view of the mountains. There was a local family feeding duck on the lake. We then strolled back to town and went into a couple of shops. This is a beautiful place.
Our next stop was Glacier bay to see the Hubbard Glacier. The captain bought us really close, and the naturalist on the boat gave a lecture through the P.A. system which was extremely interesting. This place alone made the trip worthwhile. Although we had amazing weather for the rest of the trip, this place is expectedly COLD.....Bring a coat.
Next stop, Juneau..... We had booked one of the excursions which included whale watching and a tour of the Mendenhall Glacier. We got off the boat and boarded a bus, which took us to the museum and vista. After arriving at the park-museum, we walked along a path by a stream where the salmon were running. At the end of the path there is a vista where you get a spectacular view of the glacier. We did not go into the museum, as our time there was short, and we had to be back at the bus. The bus driver kept us entertained with stories about life there as we made our way to the boat. After a short safety talk on the boat, we got underway to look for whales. This trip offered much of our money back if we didn't see any whales, so we were hopeful. There was a naturalist who lectured during the trip and I really learned a lot. About 1/2 hour in we saw our first whale!! We followed the whale until we had seen it a number of times, and then proceed to the next siting. They served hot chocolate, coffee, and snacks, which were all delicious. We saw three whales, a number of sea lions, and possibly the most the gorgeous scenery of the entire trip. You get another perceptive of the Mendenhall glacier and a couple other glaciers from the boat. I highly recommend this excursion. Back on the ship we saw more whales from the constellation deck.
We then proceeded to Ketchikan which had a frontier town feel along a backdrop of the Tsongas rain forest. We roamed the shops and saw an animal shelter giving a talk, while we watched a live Eagle and an owl that the had rescued. We were our way again through the inside passage and more beautiful scenery.
Our final destination, Victoria B.C. was to be short, as we would get there at 7 P.M. and had to be back on the ship by 12 A.M. Once off the ship we found that they had convenient shuttles to take you to town and back. The last bus leaves at 11 P.M. My girlfriend was a bit disappointed as she would have liked to have seen Bouchard Gardens, and the time seemed to short for that. We went into town and had dinner at "Milestones" restaurant. The food was good and it has a wonderful view of Victoria. We walked over to the "Empress" hotel and checked out the lobby. The ship left for Seattle at about 11:30 P.M.
Much of the food on the ship was pretty bad. We went to the restaurant the first night and I got the Salmon which did not seem fresh. The soup was inedible. Things did not improve through the trip.On a positive note, my girlfriend who has celiac and cannot eat wheat, was able to get some gluten free bread. She told me it was the best gluten free bread that she had ever eaten. Unfortunately they ran out of it three days into the cruise. Most of the crew were eager to please, and would go to great lengths to make you happy. The young man making pizza arranged to make a gluten free pie for us. It was pretty bad but we really appreciated the effort.
The dinning room menus were very ambitious. Every item sounded wonderful, but most were rather bad. You would think that they would at least have some fresh seafood, but that was not the case. They could do a lot better on the food.
After the initial room trouble,our room steward seemed to be irritated each time we called him. Although he was not pleasant about, he did manage to do everything we asked.
Alaska is a wonderful place to see, and the boat and the crew got us there. Everything was done efficiently. Lets hope they can do better with the food. If cigarette smoke bothers you, I would advise against this ship. Otherwise, if you want to see Alaska and are not very food centered, take this cruise. Less
Published 09/17/08
Cabin review: 3018
decent room, window was a nice touch. Beware of cigarette smoke.
Read Cruise Critic's Celebrity Infinity Review >>
Amazing Alaska
Lacks the luxury from its publ...
Weather was our friend - beaut...
Great itinerary, tired ship, n...
First time to Alaska on Infini...
Alaska was simply amazing
Lacks the luxury from its publ...
Great service, good food.
My first cruise ever
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One cool thing about attending the Sonoma International Film Festival is hearing the directors, writers, producers, editors and actors talk about their films. They are all kind, outgoing, and willing and grateful to share their film with the audience. We were bombarded with emails from film makers the week of the festival, inviting us to attend their films. We attended as many as we could! A few films and interviews that stuck out in our minds include Dan Jenski of ADDicted and John Beck of Harvest.
Dan's movie was the first of The Sixth Sense shorts series at The Vintage House on Friday the 13th. This short film revolved around Alex, also played by Jenski as he struggles with the disease and becomes addicted to Adderol. After the film, we found Dan outside the theatre waiting for his film to be over. We asked why he wasn't in the theatre for his movie premier, and he told us he was too nervous– it was a stigma thing. Dan was gracious to share his film with us and the festival and is looking forward to turning his short into a full length film, currently under the works.
It's nice to have a media pass to the festival because the Sebastiani Winery Barrel Room was absolutely packed for the debut of Harvest, by John Beck, covering the struggles of Sonoma wineries in the 2011 wine-grape harvest, revealing the blood, sweat and tears that go into every bottle of wine we enjoy. This film follows the middle-of-the-night picks, the terrible harvest weather, and will make viewers appreciate the wonderful wine's they drink, giving an understanding of what goes into this art. Beck looks forward to further editing this film to be shown on PBS for the world to see and enjoy as we did.
Check out our interview with Dan of ADDicted and John of Harvest. More videos to come!


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Hi I have found a yummy buttercream recipe that has shortening and a lot of fresh cream. I have to transport decorated cupcakes 1 hour away in car and they then have to sit at room temp. It will prob be between 24-30 degrees here so what I really want to know is will I make everyone sick if I use this recipe? Don't think my friends will appreciate food poisoning for their anniversary! Hope someone can help?!
If this recipe is no good is there one that can cope with heat but is still delicious!
Hi Chelle. You could use a 100% shortening + icing sugar recipe (plus flavouring of course). There is a buttercream base product called So-Lite Tropical in cake shops here that you use instead of butter. Regular bakery/cake shops use it a lot. Or you can buy a product called Bakels Whip'n'ice or some such. It is meant to be temperature safe and can sit out of the fridge once whipped, for ages.
What I would do, however, because I hate the taste of shortening, and refuse to use that fake whipped icing crap on anything I make (check out the list of 'ingredients' on the side of the carton!), is to make my usual butter, icing sugar, flavouring, cream and chocolate buttercream (ie ganache mixed into a regular buttercream). Decorate as normal. Put the cuppies in a box and into the fridge to super-chill through before you leave. They shoudl be fine in an A/C car, and then on display for several hours before eating. I would not use my meringue-based buttercream for this occasion.
Not knowing the recipe it is really hard to tell for sure - but in general most 'b'creams' made w/shortening and powdered/icing sugar is very safe to be left UNrefrigerated for 2 weeks. I rather doubt you will make anyone sick! That is unless some kid eats a dozen of those cuppies - then they will have a tummy ache


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On Food Network
Episode: Snack Attack
Art Smith - Shrimp and Heirloom Tomato Pizzette
$25 prix fixe dinner is a great deal! choose any salad, entree (excluding specials) and dessert! good for sunday monday and tuesday nights.
Pomme Frittes with cheese fondue and black pepper. Yum!
Seasonal influences appear mostly as starters and sides for entr�es (green garlic and ramps, for instance). And crispy, paper-thin ?pizzettes? (rectangular pizzas) are served.
Terrible service. Run!
So good! Great date spot, great for small groups.
Check out Socca as seen on Best Thing I Ever Ate
Check out Socca as seen on Best Thing I Ever Ate
Best antipasto platter in Chicago.
Do yourself a favor. Get the black truffle risotto while they have it. Great restaurant, great service!
Omg the PBJ is amazing. Must have dessert and more!!!


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Jul 16, 2013
Could You Have a Deviated Septum?
What is the septum? It is the wall of cartilage and bone that separates your two nostrils. When this wall becomes misaligned, it is considered to be a deviated septum. What are some signs of a deviated septum? Is it serious?...


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Why this car is my childhood hero? Because this car in my childhood hung the poster. Since my childhood I admire old vehicles, their development and especially design. For me nejlegendárnějším car then Mercedes Benz SSKL 1931. Auto, from which it began to develop other racing cars. This legend does not die. I created the exact scene in Nürburgring racing circuit in 1931 and the whole result was then transferred to a retro style. :)
choice of professionals


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24 Jul The Racing Capital of the World – The History of Indianapolis Motor Speedway
The weekend NASCAR drivers are travelling to Speedway, Indiana to race at the highest capacity stadium facility in the world; the Indianapolis Motor Speedway (IMS). This massive speedway has the capability to seat more than 400,000 people! Reaching a distance of two and a half miles long, IMS is so large that Churchill Downs, Yankee Stadium, the Rose Bowl, the Roman Colosseum and Vatican City could all fit inside the oval, which covers 253 acres! Before Sunday's Brickyard 400 race, let's take a look back at this speedway's interesting history, traditions and records.
The Indianapolis Motor Speedway was built in 1909 and was the first racetrack to be titled a speedway, making it the original speedway. Since August 19, 1909, 248 automobile races have taken place at IMS, including the Indianapolis 500 and one of NASCAR's most popular races, the Brickyard 400. The Brickyard 400's first race brought in more than 250,000 spectators, the largest crowd to date to witness a NASCAR event!
You have probably heard of the Brickyard 400, but do you know where this race's name comes from? The term "brickyard" refers to the 3.2 million bricks that were used to pave the Indianapolis Motor Speedway back in 1909. By 1961 the entire track was paved with asphalt with the exception of a one-yard strip of brick that remains at the track's start- finish line. This section of bricks is widely known today as the Yard of Bricks.
NASCAR driver Dale Jarrett began a Brickyard tradition that would soon catch on in the racing world. After winning the Brickyard 400 race in 1996, Jarrett and his crew chief Todd Parrott walked over to the start-finish line, knelt down and kissed the Yard of Bricks. This started a tradition that has carried on with NASCAR drivers ever since.
After winning the Brickyard 400's first race on August 6, 1994, rookie NASCAR driver Jeff Gordon went on to prove that his Brickyard victory was not just beginner's luck. Gordon holds the track's record for the most victories (four), most top five finishes (10) and most top 10 finishes (13).
Indianapolis isn't called the racing capital of the world for nothing! The speedway got national recognition for its historical significance in 1975 when it was placed on the National Register of Historic Places and was also designated as a National Historic Landmark in 1987. It remains the only landmark to be affiliated with automotive racing. Now that you know more about the Indianapolis Motor Speedway and the Brickyard 400, you can get ready for some high speed action at this weekend's race and help us cheer on 5-hour ENERGY®'s own Clint Bowyer!