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The nestin-expressing and non-expressing neurons in rat basal forebrain display different electrophysiological properties and project to hippocampus
Nestin-immunoreactive (nestin-ir) neurons have been identified in the medial septal/diagonal band complex (MS/DBB) of adult rat and human, but the significance of nestin expression in functional neurons is not clear. This study investigated electrophysiological properties and neurochemical phenotypes of nestin-expressing (nestin+) neurons using whole-cell recording combined with single-cell RT-PCR to explore the significance of nestin expression in functional MS/DBB neurons. The retrograde labelling and immunofluorescence were used to investigate the nestin+ neuron related circuit in the septo-hippocampal pathway.
The results of single-cell RT-PCR showed that 87.5% (35/40) of nestin+ cells expressed choline acetyltransferase mRNA (ChAT+), only 44.3% (35/79) of ChAT+ cells expressed nestin mRNA. Furthermore, none of the nestin+ cells expressed glutamic acid decarboxylases 67 (GAD67) or vesicular glutamate transporters (VGLUT) mRNA. All of the recorded nestin+ cells were excitable and demonstrated slow-firing properties, which were distinctive from those of GAD67 or VGLUT mRNA-positive neurons. These results show that the MS/DBB cholinergic neurons could be divided into nestin-expressing cholinergic neurons (NEChs) and nestin non-expressing cholinergic neurons (NNChs). Interestingly, NEChs had higher excitability and received stronger spontaneous excitatory synaptic inputs than NNChs. Retrograde labelling combined with choline acetyltransferase and nestin immunofluorescence showed that both of the NEChs and NNChs projected to hippocampus.
These results suggest that there are two parallel cholinergic septo-hippocampal pathways that may have different functions. The significance of nestin expressing in functional neurons has been discussed.
Medial septal/diagonal band complex (MS/DBB) is a highly heterogeneous region with different types of neurons and implicated in various functions such as arousal, sensory processing, motivation, emotion, learning and memory
There are four classes of neurons in the MS/DBB distinguished by electrophysiological characteristics
Nestin is an intermediate filament protein expressed transiently by neural progenitor cells and reactivated glial cells
Chemical phenotypes of MS/DBB neurons identified by sc-RT-PCR
A total of 106 Medial Septal/Diagonal Band Complex (MS/DBB) neurons were electrophysiologically recorded and their chemical phenotypes were identified by multiplex sc-RT-PCR. The results showed that the mRNAs encoding nestin, ChAT, glutamic acid decarboxylases 67 (GAD67), vesicular glutamate transporters 1 or 2 (VGLUT1 or VGLUT2) could be reversely transcribed and amplified from the harvested cytoplasm. Automatic sequencing confirmed that each PCR product is from the target cDNA. The MS/DBB neurons studied in our experiment were comprised of 79 ChAT mRNA-positive neurons (ChAT+) that are cholinergic neurons, 13 GAD67 mRNA-positive neurons (GAD67+) that are GABAergic neurons. There were 11 neurons co-expressing ChAT mRNA and GAD67 mRNA, of which, 6 were categorized as cholinergic neuron and 5 as GABAergic neuron according to their electrophysiological properties. Nine neurons solely expressed VGLUT1 mRNA or/and VGLUT2 mRNAs but not ChAT mRNA or GAD67 mRNAs, which confirms the identification of glutamatergic neurons
The intrinsic membrane properties of nestin mRNA+ neurons
Eighty-seven neurons were assessed for their electrophysiological profile. This assessment identified 69 slow-firing neurons, 5 cluster-firing neurons and 13 fast-firing neurons. Although some neurons presented rebound action potentials following a hyperpolarization current injection, we did not find any typical burst-firing neuron in our experiment. All of the recorded nestin+ cells were excitable, with typical electrophysiological characteristics of functional neurons. In voltage-clamp mode, when depolarized from -60 mV, typical neuronal whole-cell currents (comprised of large inward Na+ current and outward K+ current) could be elicited (Figure
Figure 1
Nestin mRNA-positive cells in MS/DBB are functional neurons
Nestin mRNA-positive cells in MS/DBB are functional neurons. A. Agarose gel analysis of the sc-RT-PCR products obtained from a single MS/DBB cell. The only PCR-generated fragment was nestin. B. Whole cell current of the same cell depolarized from -60 mV to 10 mV in voltage-clamp mode showed characteristic of functional neurons. C. The cell shows typical neural action potential when depolarized to the threshold potential. D. A sustained depolarizing current (1000 ms) elicits a train of repetitive action potentials. The membrane responses were elicited using positive currents of 0.2 nA from -60 mV in panel C and D.
Table 1
Electrophysiological properties of chemically identified neurons in the rat medial septal/diagonal band complex
nestin- & ChAT+
number of neurons
Cm (pf)
55.76 ± 3.05 b
49.60 ± 2.37 b
76.99 ± 9.60
43.58 ± 3.92 b
Rm (MΩ)
400.56 ± 26.86
387.26 ± 27.27
289.63 ± 44.25
570.07 ± 120.25
Tau (ms)
0.85 ± 0.07
0.70 ± 0.05a
1.27 ± 0.18
0.64 ± 0.06 a
RMP (mV)
-56.73 ± 1.05
-59.75 ± 1.14
-55.62 ± 1.91
-58.00 ± 3.47
AHP duration (ms)
223.77 ± 12.08 b
230.52 ± 8.48 b
165.23 ± 7.04
238.17 ± 16.97 a
AHP amplitude (mV)
4.49 ± 0.21 b
5.02 ± 0.35 b
3.27 ± 0.25
4.80 ± 0.28 b
AP amplitude (mV)
99.04 ± 1.37 a
97.39 ± 1.22 b
104.97 ± 2.22
98.85 ± 2.82
Spike half width (ms)
0.88 ± 0.04 b
0.94 ± 0.03 b
0.67 ± 0.06
0.99 ± 0.05 b
Spike width (ms)
2.46 ± 0.09 b
2.66 ± 0.06 b
2.02 ± 0.14
2.78 ± 0.14 b
rise time (ms)
0.30 ± 0.01 b
0.32 ± 0.01 b
0.23 ± 0.02
0.32 ± 0.01 b
rise slope (mV/ms)
121.91 ± 8.30
105.85 ± 4.99 a
196.45 ± 24.41
106.31 ± 8.06 a
decay time (ms)
0.69 ± 0.03 b
0.73 ± 0.03b
0.51 ± 0.05
0.82 ± 0.06 b
decay slope (mV/ms)
-49.57 ± 3.21 a
-43.69 ± 1.81 b
-83.06 ± 9.41
-38.98 ± 2.55 b
MF (Hz)
8.39 ± 0.45 b
8.19 ± 0.49 b
18.38 ± 1.73
9.22 ± 0.97 b
FMAX (Hz)
20.73 ± 2.07 a
18.86 ± 1.77 b
28.99 ± 1.93
19.83 ± 0.82 b
FSTEADY (Hz)
6.86 ± 0.53 b
6.67 ± 0.35 b
17.23 ± 1.66
7.01 ± 0.49 b
spike adaptation
0.55 ± 0.05 b
0.56 ± 0.04 b
0.28 ± 0.06
0.56 ± 0.11 a
depolarizing sag (mV)
2.74 ± 0.62 b
1.81 ± 0.32 b
16.81 ± 0.77
2.22 ± 0.56 b
-12.90 ± 2.57 b
-4.00 ± 2.17 b
-186.82 ± 15.73
-3.49 ± 1.86 b, c
Nestin+, nestin mRNA-positive neuron (including nestin mRNA-positive & ChAT mRNA-negative neurons and nestin mRNA-positive & ChAT mRNA-positive neurons); Nestin- & ChAT+, nestin mRNA-negative and choline acetyl transferase mRNA-positive neuron; GAD67+, glutamic acid decarboxylases 67 mRNA-positive neuron; VGLUT+, vesicular glutamate transporter 1 or/and 2 mRNA-positive neuron. Cm, membrane capacitance; Rm, membrane resistance; Tau, time constant; RMP, resting membrane potential; threshold, action potential threshold; AHP, afterhyperpolarization; MF, Mean firing rate; FMAX, maximal firing frequency; FSTEADY, steady firing frequency.
Figure 2
Comparison of action potential properties of MS/DBB neurons
Comparison of action potential properties of MS/DBB neurons. A. Representative action potentials from all cell types (superimposed) show the differences in spike shape and width among the four cell types. GAD67+ neurons have the narrowest action potentials, where as VGLUT+ neurons have the broadest action potentials. B. Histogram of action potential amplitude of all cell types. C. Histogram of spike parameters of all cell types. *
Figure 3
Electrophysiological properties of nestin+ and/or ChAT+ neurons
Electrophysiological properties of nestin+ and/or ChAT+ neurons. A1-F1: Nestin+ & ChAT- neurons, A2-F2: Nestin+ & ChAT+ neurons, A3-F3: Nestin- & ChAT+ neurons. A1-3: Agarose gel analysis of the sc-RT-PCR products to identify chemical phenotypes of recorded neurons. B1-3 and C1-3: In current-clamp mode, membrane responses of the same MS/DBB neuron to depolarizing current pulses (0.2 nA) applied from membrane potential of -60 mV (B1-3) or -80 mV (C1-3). All neurons display slow-firing activity. D1-3: Injection of a hyperpolarizing current pulse from -60 mV, no depolarizing sag and rebound firing were found in each kind of neuron. E1-3: Hyperpolarizing voltage steps applied from a holding potential of -50 mV showed absence of conspicuous
Figure 4
Electrophysiological properties of GAD67+ and VGLUT+ neurons
Electrophysiological properties of GAD67+ and VGLUT+ neurons. A1-F1: GAD67+ neuron presents electrophysiological properties of fast-firing neuron. A2-F2: VGLUT+ neuron presents electrophysiological properties of cluster-firing neuron. A1-2: Agarose gel analysis of the sc-RT-PCR products to identify chemical phenotypes of recorded neurons. B1 and C1: Membrane responses of the same MS/DBB neuron to injection of depolarizing current pulses applied from a membrane potential of -60 mV (B1) or -80 mV (C1). GAD67+ neuron displays fast-firing activity. B2 and C2: Membrane responses of the VGLUT+ neuron to injection of depolarizing current pulses (0.2 nA) applied from a membrane potential of -60 mV, prolonged depolarization current (4s in duration) elicits cluster-firing separated by subthreshold membrane oscillations (C2). D1-2: In current-clamp mode, the response to injections of hyperpolarizing current pulse from -60 mV, note profound depolarizing sag in GAD67+ neuron, but no rebound firing action potential (D1). E1-2: Currents recorded in voltage-clamp mode evoked by a series of hyperpolarizing voltage steps applied from a holding potential of -50 mV. Notice the profound depolarizing amplitude inward current in GAD67+ neuron, as shown by the differences between the amplitudes of the instantaneous current (filled circle) and steady-state current (open circle). F1-2: Instantaneous and steady-state I-V plots derived from the data in E1-2.
Figure 5
Comparison of RMP and
Comparison of RMP and
Nestin+ neurons (including those neurons co-expressing ChAT mRNA) had a mean fire rate (MF) of 8.39 ± 0.45 Hz, maximal firing frequency (FMAX) of 20.73 ± 2.07 Hz, and steady firing frequency (FSTEADY) of 6.86 ± 0.53 Hz. These neurons also had broad action potentials (spike width 2.46 ± 0.09 ms) and large afterhyperpolarization (duration 223.77 ± 12.08 ms, amplitude 4.49 ± 0.21 mV), which were similar to nestin- & ChAT+ neurons (
Interestingly, while further analyzed the electrophysiological properties of nestin+ & ChAT-, nestin+ & ChAT+ and nestin+ & ChAT- neurons, we found that the
Excitatory postsynaptic currents recorded from nestin+ & ChAT+ neurons and nestin- & ChAT+ neurons
In this section, nineteen neurons, which contained 12 nestin- & ChAT+ neurons and 7 nestin+ & ChAT+ neurons, were recorded, and 3993 sEPSCs events and 3570 mEPSCs events were analyzed in total. The addition of 10 μM 6-cyano-7-nitroquinoxaline-2, 3-dione (CNQX, a non-NMDA glutamate receptor antagonist) abolished all synaptic events, indicating the involvement of non-NMDA glutamate receptors (data not shown).
The sEPSCs amplitude (28.45 ± 1.78 pA) of nestin+ & ChAT+ neurons was significantly larger than that of the nestin- & ChAT+ neurons (22.91 ± 1.05 pA) (student's
Figure 6
Comparison of sEPSCs of MS/DBB nestin- and nestin+ cholinergic neurons
Comparison of sEPSCs of MS/DBB nestin- and nestin+ cholinergic neurons. A. Consecutive traces of sEPSCs recorded from MS/DBB nestin- and nestin+ cholinergic neurons. B. Average sEPSCs of nestin- and nestin+ cholinergic neurons. C. Comparison of the sEPSCs frequencies of MS/DBB nestin- and nestin+ cholinergic neurons. D. Comparison of the sEPSCs amplitudes of MS/DBB nestin- and nestin+ cholinergic neurons (*:
In order to further explore the mechanism of the different sEPSCs between the nestin- & ChAT+ and nestin+ & ChAT+ neurons, mEPSCs of both kinds of neurons were studied. The independent samples student's
Figure 7
Comparison of mEPSCs of MS/DBB nestin- and nestin+ cholinergic neurons
Comparison of mEPSCs of MS/DBB nestin- and nestin+ cholinergic neurons. A. Consecutive traces recorded from MS/DBB nestin- and nestin+ cholinergic neurons. B. Average mEPSCs of nestin- and nestin+ cholinergic neurons. C. Comparison of the mEPSCs frequencies of MS/DBB nestin- and nestin+ cholinergic neurons. D. Comparison of the mEPSCs amplitudes of MS/DBB nestin- and nestin+ cholinergic neurons (**:
The paired samples student's
Figure 8
Comparison of sEPSCs and mEPSCs of nestin- and nestin+ cholinergic neurons in MS/DBB
Comparison of sEPSCs and mEPSCs of nestin- and nestin+ cholinergic neurons in MS/DBB. A. Comparison of the frequencies of sEPSCs and mEPSCs of nestin- and nestin+ cholinergic neurons. B. Comparison of the amplitude of sEPSCs and mEPSCs of nestin- and nestin+ cholinergic neurons. C. Multiplicity of nestin- and nestin+ cholinergic neurons. D. sEPSCs/mEPSCs frequency ratio of nestin- and nestin+ cholinergic neurons. *:
Immunofluorescence study of the biocytin-filled neurons
Twenty-eight neurons were successfully filled with biocytin and visualized by Rhodamine Red-X. Cell bodies were particularly well-labelled, allowing us to determine their position relative to the MS/DBB. Biocytin-filled neurons were bipolar or multipolar and gave off two or three primary dendrites that subsequently bifurcated to the adjacent areas. The axons originated from the soma or proximal end of a primary dendrite. No evidence of axon collaterals was found in our slices. Of the 28 biocytin-filled neurons, 22 were ChAT-immunoreactive (ChAT-ir) neurons, among which 45.45% (10/22) were nestin-immunoreactive (nestin-ir) neurons. Eleven out of the 28 biocytin-filled neurons were nestin-ir neurons, 90.91% (10/11) of which were also ChAT-immunoreactive (Figure
Figure 9
Triple immunofluorescent study of biocytin-filled neuron
Triple immunofluorescent study of biocytin-filled neuron. A. Biocytin-filled neuron was visualized by rhodamine red-X-conjugated streptavidin. B and C showed ChAT- and nestin-immunoreactive neurons visualized by cy2 (blue) and alexa 405 (green) respectively. D. Image merged from A, B and C. The red arrow pointed to the cell double labelled by the nestin and ChAT antibodies and filled with biocytin by whole-cell patch clamp recording. The white arrow pointed to the cell double labelled by ChAT and nestin antibodies. The blue arrow pointed to a neuron that only expressed ChAT. Because the brain slice was made from neonatal rat and did not perfuse transcardially before slice preparation, there were some epithelium lining blood vessels labelled by nestin monoclone antibody in C and D. Scale bar was 20 μm.
Retrograde tracing of fast blue from the CA1 area of hippocampus
Examination of serial section of the basal forebrain region 5 days after injection of the fast blue revealed that the blue colour fluorescence could be visualized clearly. The fast blue labelled somas were seen throughout the entire MS/DBB region. Histological examination of the MS/DBB area after labelling revealed striking intense signals in the cell body, however, the neuritis were difficult to distinguish from background. In order to define the anatomical circuits of the nestin+ and nestin- cholinergic projection to the hippocampus, we evaluated the percentage of ChAT and nestin immunoreactivity among the fast blue-labelled neurons in the MS/DBB region after fast blue intra-hippocampus instillation. The nestin and ChAT immunoreactive cells were clearly labelled by green and red colour fluorescence specifically. In order to find the ratio of the nestin+ or nestin- cholinergic neurons projection to the hippocampus, the double or triple fluorescence of combined immunohistochemistry and retrograde labelling were carefully measured. Approximately 20.40% of the fast blue-labelled neurons in the MS/DBB area were ChAT-immunoreactive. In which, 48.04% were nestin-expressing neurons, and 51.96% nestin non-expressing cholinergic neurons (Figure
Figure 10
Retrograde labelling demonstrates that the nestin+ and nestin- cholinergic neurons projected to hippocampus
Retrograde labelling demonstrates that the nestin+ and nestin- cholinergic neurons projected to hippocampus. (A) Photomicrograph demonstrating the deposition of fast blue dye throughout the entire MS/DBB area and the location of ChAT+ and nestin+ neurons. (B) The ChAT+ neurons in MS/DBB area. (C) The nestin+ neurons in the MS/DBB area. (D) The fast blue transported from the hippocampus was localized in the neurons of the MS/DBB area. (E) The double immunostaining of ChAT+ neurons and nestin+ neurons, arrows point to the double labelling neurons (nestin+ cholinergic neurons). (F) Photomicrograph of neurons labelled by retrograde tracing of fast blue from the hippocampus and nestin+/nestin- cholinergic neurons. Arrows and arrowheads point to the nestin+ and nestin- cholinergic neurons labelled with fast blue. Scale bar: 50 μm in A; 50 μm in B-F.
The main findings of this study were as follows: the electrophysiologically recorded cells expressing nestin mRNA in the MS/DBB are functional neurons; the majority of nestin+ neurons are cholinergic neurons rather than GABAergic or glutamatergic neurons; nestin+ & ChAT+ neurons are more excitable and received stronger excitatory synaptic afferent currents than those of the nestin- & ChAT+ neurons. In addition, the fast blue retrograde labelling experiment demonstrates that both the nestin+ and nestin- cholinergic neurons sent projections to hippocampus.
The recorded cells expressing nestin in MS/DBB are functional neurons
Using whole-cell recording combined with sc-RT-PCR, our experiment demonstrated that all of the electrophysiologically recorded cells expressing nestin mRNA in MS/DBB were excitable. Whole cell currents of functional neurons could be elicited from these neurons in voltage-clamp mode; typical neural action potential was observed in response to a short period of depolarization, and repetitive action potentials could be elicited when sustained positive current was applied in current-clamp mode. Due to their typical intrinsic membrane properties recorded here, we concluded that these cells are functional neurons rather than stem cells or glial cells, since neural stem cells and glial cells could not be excited to produce neural action potential. For the first time, these data confirmed that nestin+ cells in the MS/DBB are functional neurons by the joint evidence of mRNA expression and electrophysiological properties from whole-cell recordings.
Most of the nestin+ neurons in MS/DBB were cholinergic neurons
Sc-RT-PCR results revealed 87.5% of nestin+ neurons expressed ChAT mRNA, and about 44.3% of ChAT+ neurons expressed nestin mRNA. However, no nestin+ neurons expressed GAD67 mRNA or VGLUT mRNA. These results were further confirmed by the nestin and ChAT immunofluorescent labelling of biocytin filled neurons and fast blue retrograded labelled neurons under the laser confocal microscope, which confirmed that nearly one half of the ChAT-ir neurons were nestin-ir neurons. There were a few nestin+ neurons that did not express ChAT+, GAD67 or VGLUT mRNA. This may stem from limit of methodology, or there were a new class of neurons in MS/DBB.
Taken together, these observations provide new evidence that the majority of nestin+ neurons in MS/DBB were cholinergic neurons rather than GABAergic or glutamatergic neurons. In other words, cholinergic neurons in the MS/DBB could be subdivided into nestin-expressing cholinergic neurons (NEChs) and nestin non-expressing cholinergic neurons (NNChs). This result is partially inconsistent with the previous reports, in which about 35% of nestin-ir neurons were ChAT-ir
NEChs and NNChs had different electrophysiological properties
NEChs and NNChs project parallelly to hippocampus
MS/DBB is one of the most important inputs to the hippocampal neurons
In our experiment, we have demonstrated that nestin+ neurons are a subtype of basal forebrain cholinergic neurons using single-cell RT-PCR, immunohistochemistry and electrophysiological property analysis. Therefore, basal forebrain cholinergic neurons could be divided into two groups according to whether expressing nestin and their electrophysiological properties. Retrograde labelling combined with ChAT and nestin immunofluorescence suggested that both of the nestin+ and the nestin- cholinergic neurons project to the hippocampus. Therefore, we concluded that there are two parallel septo-hippocampal cholinergic pathways. One pathway originates from MS/DBB nestin-expressing cholinergic neurons (NEChs), and the other pathway originates from nestin non-expressing cholinergic neurons (NNChs). Because the NEChs and NNChs had different intrinsic electrophysiological properties and received distinct excitatory synaptic inputs, they may have different functions in maintaining the electrical activities in the hippocampus, which is worthy of further study.
Nestin is expressed transiently by neural progenitor cells and reactivated glial cells
In conclusion, we studied the electrophysiological properties of the novel nestin+ neurons in the MS/DBB, and demonstrated that most of nestin+ neurons are functional cholinergic neurons. We also provided evidence that the NEChs had higher excitability and received stronger spontaneous excitatory synaptic inputs than those of the NNChs. Then we demonstrated that both of the NEChs and NNChs projected to the hippocampus. The different electrophysiological properties of NEChs and NNChs and common neural circuits to hippocampus suggested that there are two parallel septo-hippocampal cholinergic pathways that may have different functions. Whether nestin expression affects the cholinergic neurons' properties require further study by manipulation of gene expression. These results will not only facilitate our understanding of the structures and biological function of basal forebrain, the mechanism of learning and memory, the ageing process and the pathology of AD, but may also provide a new insight for AD treatment.
Ethics Statement
All experiments were approved by Institutional Animal Care and Use Committee of Sun Yat-sen University. All work was carried out in accordance with the National Institute of Health Guide for the Care and Use of Laboratory Animals (NIH Publications No. 80-23) revised 1996. Every effort was made to minimize the animals used and their suffering.
Brain slice preparation
Brain slices containing the MS/DBB were prepared from 40 Sprague-Dawley rats (14-21 days postnatal) of either sex as previously described
Whole-cell recordings
Patch pipettes were made from diethyl pyrocarbonate (DEPC, Amresco, USA) processed and heat-sterilized borosilicate glass capillary tubing containing a filament (OD: 1.5 mm, ID: 0.86 mm) by Sutter P-97 horizontal puller (Sutter Instruments Co., Novato, USA). The pipette filled with 5μl internal solution had 3-6 Mohm pipette resistance. The internal solution was prepared with nuclease-free water containing (in mM): 144 potassium gluconate, 3 MgCl2, 10 Hepes, 0.2 EGTA, 2 K2-ATP, 0.3 Na3-GTP, pH 7.2 (285-295 mOsm). Whole-cell recordings in voltage-clamp and current-clamp modes were performed at 26 ± 0.5°C using an Axon Multi Clamp-700B amplifiers and a Digidata 1322A analog-to-digital converter (Axon Instruments Inc, USA). The output signal was continuously filtered at 3 kHz, digitized at a sampling rate of 20 kHz, and stored in a computer for off-line analysis using pClamp 9.2 software (Axon instruments Inc, USA) or Mini Analysis 6.0.3 (Synaptosoft, Leonia, NJ, USA) software.
Intrinsic membrane properties analysis
All electrophysiological recordings were made from the medial septum and the vertical limb of the diagonal band of Broca. The resting membrane potential of each neuron was measured in current-clamp mode (I = 0) just after passing in whole cell configuration. All membrane potentials were corrected for junction potential (about -15 mV). The experiment was continued only when the resting membrane potential was more negative than -45 mV, spikes overshot 0 mV and the series resistance was less than 30 Mohm
Figure 11
Measurement of action potential parameters and firing patterns
Measurement of action potential parameters and firing patterns. A, B: Measurement of action potential parameters. 1) Resting membrane potential (mV); 2) Action potential threshold (mV); 3) Action potential amplitude (mV); 4) After hyperpolarization (AHP) amplitude (mV); 5) AHP duration (ms); 6,)Spike rise time (ms); 7) Spike decay time (ms); 8) Spike duration; 9), Spike half width (ms). C, D: Measurement of parameters of firing properties of a medial septal and diagonal band complex neuron subjected to a depolarizing current pulse: time interval between: 1, the first two action potentials; 2, the last two action potentials. Reciprocal of time intervals gives maximum firing frequency (FMAX) and steady firing frequency (FSTEADY). Scale bars: 20 mV, 50 ms in A; 20 mV, 0.5 ms in B; 20 mV, 100 ms in C.
Firing patterns were analyzed in response to series of 1-4 s depolarizing current pulses applied from - 60 mV, and repeated from a membrane potential of -80 mV. Firing frequencies were determined for each neuron depolarized from -60 mV using a current pulse of 100-300 pA. Maximal firing frequency (FMAX) and steady firing frequency (FSTEADY) were calculated from the time interval between the first two spikes or the last two spikes respectively in a train of action potentials evoked by a 1 s depolarizing current pulse, and mean firing rate was calculated from the number of spikes evoked by a 1 s depolarizing current pulse (Figure
The voltage-dependent inward rectification (depolarizing sag) was determined by application of a series of 4 s hyperpolarizing current from -60 mV, and its amplitude was measured for the current pulse inducing an initial hyperpolarization to - 95 mV. The hyperpolarization activated current (
Analysis of the excitatory synaptic input current properties
The selected neurons were clamped at -70 mV. The internal solution was the same as that used in the experiment for intrinsic membrane properties study. To study spontaneous excitatory postsynaptic currents (sEPSCs), bicuculline (10 μM) was added to the perfusing solution to block GABAA-mediated synaptic transmission. Tetrodotoxin (TTX, 1 μM) was added to the bicuculline-containing solution to obtain mEPSCs. The output signal was continuously filtered at 3 kHz, digitized at a sampling rate of 20 kHz, and stored in a computer for off-line analysis using Mini Analysis 6.0.3 (Synaptosoft, Leonia, NJ, USA) software. The excitatory postsynaptic currents analysis was based on 180 s gap free recordings. The detection threshold was set at twice the baseline noise. The fact that no false events would be identified was confirmed by visual inspection for each synaptic current. The cells were rejected if the access resistance changed more than 20% during the experiment. The addition of 10 μM 6-cyano-7-nitroquinoxaline-2, 3-dione (CNQX, a non-NMDA glutamate receptor antagonist) abolished all synaptic events, indicating the involvement of non-NMDA glutamate receptors (data not shown).
To compare the synaptic connectivity, we calculated multiplicity index, a common parameter used as a maturation index in synaptic developmental studies, defined as the number of release sites per presynaptic neuron could release neurotransmitter simultaneously onto the recorded cell
Where Vb and Vq are the mean frequency of sEPSCs and mEPSCs; b and q are the mean amplitude of sEPSCs and mEPSCs.
Cytoplasm harvest and reverse transcription
Cytoplasm harvest and reverse transcription (RT) were performed as previously described
Multiplex PCR
The cDNAs for ChAT, glutamic acid decarboxylases 67 (GAD67), vesicular glutamate transporters 1 and 2 (VGLUT1 and VGLUT2), and nestin were amplified with a two-step PCR protocol, slightly modified from that described by Sotty et al. (2003) and Puma et al. (2001). Target cDNAs were amplified simultaneously as a multiplex PCR in the first round and amplified individually in the second round PCR with nested primers. The first round PCR reaction was performed in a Mastercycler (Eppendorf, Germany) with a final volume of 100 μl containing the 21 μl RT reaction, 1× PCR Buffer (50 mM KCl, 10 mM Tris-HCl, 1.5 mM MgCl2, pH 8.3), 50 μM of each of the dNTPs, 10 p mol of each selected primer and 2.5 U TaKaRa Ex TaqTM HS polymerase (Takara Biotechnology, Dalian, China). The PCR amplification protocol was as follows: 2 min at 94°C; 20 cycles: 30 s at 94°C, 30 s at 60°C, 35 s at 72°C; 5 min at 72°C. Second-round PCRs were performed individually in reaction volume of 25 μl, using 2 μl of the first round PCR product and specific primer pair by 35 PCR cycles under similar conditions as first round PCR but the concentration of dNTPs was 200 mM. Five microliters of each last-round PCR product was run on a 1.5% TEA agarose gel stained with ethidium bromide, using a 100 bp DNA ladder as molecular weight marker. Some PCR products had been sent to professional company (Beijing AuGCT biotechnology Co., ltd, Beijing, China) for automatic sequencing to determine their specificity. Primers used in the experiment are shown in Table
Table 2
Primer sequences used in the experiment
Accession no.a
primer sequence
Start position b
Product sizec
(first round)
524 bp
(first round)
401 bp
(first round)
311 bp
(first round)
419 bp
(first round)
434 bp
(second round)
324 bp
(second round)
137 bp
(second round)
211 bp
(second round)
264 bp
(second round)
185 bp
ChAT, choline acetyltransferase; GAD, glutamic acid decarboxylases; VGLUT, vesicular glutamate transporter. a The GenBank accession number for the cDNA sequence used in primer design; b The cDNA position of the 5'' end of primer; c Expected PCR product length.
Immunohistochemistry and laser confocal scanning of recorded neurons
For subsequent immunofluorescence study of the recorded cells, biocytin (0.2%) was added to internal solution. In the cells from which recordings had been made, biocytin, nestin and ChAT were visualized by triple fluorescence: Rhodamine Red was the fluorescent label for biocytin, cy2 for nestin-ir and Alexa 405 for ChAT-ir. After whole-cell recording, the brain slices were fixed by immersion in 4% paraformaldehyde in 0.1 M phosphate buffer, pH 7.2, at 4°C for l-2 h, washed in 0.1 M phosphate buffer, and cryoprotected by immersion in 30% buffered sucrose at 4°C for several hours. The following day, serial sections at 50 μm thickness were cut on a cryostat, and collected in 0.01 M phosphate-buffered saline (PBS). Sections were washed (3 × 10 min) in 0.01 M PBS followed by incubation in 1% bovine serum albumin (containing 0.3% Triton X-100 in PBS) for 30 min to prevent non-specific conjugate binding. Sections were incubated with monoclonal mouse anti-nestin antibody (1:800, Rat-401, Pharmingen) and polyclonal rabbit anti-ChAT antibody (1:1000, Chemicon) for 2 h at 37°C and then for 14 h at 4°C. After a 3 × 10 min rinse in PBS, the sections were incubated in a cocktail containing Rhodamine Red X-conjugated streptavidin (1:5000, Jackson Immuno Research Laboratories), a cy2-conjugated goat anti-mouse antibody (1:200, Jackson Immuno Research Laboratories) and a Alexa 405-conjugated goat anti-rabbit antibody (2 μg/ml, Invitrogen) for 2 h at room temperature
In control experiments, the primary antibodies were omitted and, as expected, this resulted in the absence of any cellular labelling. Additional controls were performed by switching the fluorochromated immunoreagents related to the markers visualized by triple fluorescence labelling procedures which resulted in identical staining patterns.
All sections were then analyzed with the LSM 510 Meta (Zeiss). Nestin-ir was observed with the blue FITC filter; ChAT-ir was viewed with the DAPI filter; biocytin-filled cell was viewed with the Rhodamine Red filter. Confocal images were scanned, edited, reconstructed and measured using LSM 510 Meta software (Zeiss).
Analyses of the projections of nestin+ and nestin- cholinergic neurons
To demonstrate whether the nestin+ and nestin- cholinergic neurons projected to hippocampus, retrograde labelling combined with double immunostaining protocols were used. Four rats were instilled with 0.5 μl of 3% fast blue dye into the CA1 area (AP-3.8, L 1.5, H 3.0) of the hippocampus. After a 5-d transport period, the experimental animals were sacrificed, immunofluorescence of nestin and ChAT were performed with a standard protocols. Rhodamine Red was the fluorescent label for ChAT-ir, cy2 for nestin-ir neurons
Statistical analysis
All statistical analyses were performed with SPSS 11.5 for Windows. Data were analyzed statistically using either the student's
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
JHZ designed and carried out the study, performed the statistical analysis, and drafted the manuscript. HYG participated in the design of the study, performed the statistical analysis, and helped to draft the manuscript. ZBY conceived the study, and participated in its design and coordination, and helped to draft the manuscript. JTZ carried out the molecular genetic studies, participated in the sequence alignment. KHG participated in the immunohistochemistry and neuronal projection study. DPL participated in the design of the study and performed the statistical analysis. TMG participated in design and coordination of the study, and helped to draft the manuscript. All authors read and approved the final manuscript.
Acknowledgements and Funding
We gratefully acknowledge the assistance of Monique David in manuscript revision, Yongjun Chen, Pu Wang, QunFang Yuan and Yao Xie for their help in experimental techniques. This work was supported by National Nature Science Foundation of China (No. 30700436, 31160221), Doctoral Program of Guangdong Natural Science Foundation (No. S2011040004372). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Septal networks: relevance to theta rhythm, epilepsy and Alzheimer's disease
Individual differences in aging: behavioral and neural analyses
Mild cognitive impairment and the cholinergic hypothesis: a very different take on recent data
Distinct electrophysiological properties of glutamatergic, cholinergic and GABAergic rat septohippocampal neurons: novel implications for hippocampal rhythmicity
Characterization of medial septal glutamatergic neurons and their projection to the hippocampus
Evidence for glutamate, in addition to acetylcholine and GABA, neurotransmitter synthesis in basal forebrain neurons projecting to the entorhinal cortex
Parvalbumin, calbindin, or calretinin in cortically projecting and GABAergic, cholinergic, or glutamatergic basal forebrain neurons of the rat
Evidence for a distinct group of nestin-immunoreactive neurons within the basal forebrain of adult rats
Cholinergic and peptidergic projections from the medial septum and the nucleus of the diagonal band of Broca to dorsal hippocampus, cingulate cortex and olfactory bulb: a combined wheatgerm agglutinin-apohorseradish peroxidase-gold immunohistochemical study
Multiple output pathways of the basal forebrain: organization, chemical heterogeneity, and roles in vigilance
The significance of the cholinergic system in the brain during aging and in Alzheimer's disease
Membrane properties of cell types within guinea pig basal forebrain nuclei in vitro
Electrophysiological characteristics of cholinergic and non-cholinergic neurons in the rat medial septum-diagonal band complex
Role of the afterhyperpolarization in control of discharge properties of septal cholinergic neurons in vitro
Muscarinic tone sustains impulse flow in the septohippocampal GABA but not cholinergic pathway: implications for learning and memory
Glutamatergic neurons of the mouse medial septum and diagonal band of Broca synaptically drive hippocampal pyramidal cells: relevance for hippocampal theta rhythm
Nestin in central nervous system cells
Nestin immunoreactivity of Purkinje cells in Creutzfeldt-Jakob disease
Distribution of nestin immunoreactivity in the normal adult human forebrain
A cluster of nestin immunoreactive neurons exist in basal forebrain of adult rats
Age-related alterations of Nestin-immunoreactive neurons in rat basal forebrain with aged memory deficit
Stereological estimates of the basal forebrain cell population in the rat, including neurons containing choline acetyltransferase, glutamic acid decarboxylase or phosphate-activated glutaminase and colocalizing vesicular glutamate transporters
TRPV1 antagonist, SB-366791, inhibits glutamatergic synaptic transmission in rat spinal dorsal horn following peripheral inflammation
Somatostatin presynaptically inhibits both GABA and glutamate release onto rat basal forebrain cholinergic neurons
Hippocampal theta rhythm is reduced by suppression of the H-current in septohippocampal GABAergic neurons
An analysis of the origins of the cholinergic and noncholinergic septal projections to the hippocampal formation of the rat
Cholinergic and non-cholinergic septo-hippocampal projections: a double-label horseradish peroxidase-acetylcholinesterase study in the rabbit
Effects of systemically administered NT-3 on sensory neuron loss and nestin expression following axotomy
Time course of changes in cholinergic and neurotrophin-related markers after infusion of colchicine into the basal forebrain
The Effect of Colchicine on the Nestin - IR Neurons in the Rat Basal Forebrain
Deprivation of endogenous brain-derived neurotrophic factor results in impairment of spatial learning and memory in adult rats
Perineuronal nets ensheath fast spiking, parvalbumin-immunoreactive neurons in the medial septum/diagonal band complex
Electrophysiological and morphological heterogeneity of slow firing neurons in medial septal/diagonal band complex as revealed by cluster analysis
Morphology of local axon collaterals of electrophysiologically characterised neurons in the rat medial septal/ diagonal band complex
Developmental increase of asynchronic glutamate release from hippocampal synapses in mutant taiep rat
Development of excitatory circuitry in the hippocampus
Early establishment of multiple release site connectivity between interneurons and pyramidal neurons in the developing hippocampus
AMPA receptor subunits expressed by single Purkinje cells
Molecular and physiological diversity of cortical nonpyramidal cells
Sodium current density correlates with expression of specific alternatively spliced sodium channel mRNAs in single neurons
Cell-specific regulation of agrin RNA splicing in the chick ciliary ganglion
Triple fluorescence labelling of neuronal, glial and vascular markers revealing pathological alterations in various animal models


text:
1963 Hudson Street, Muskegon 49441
See more homes for sale in Muskegon, MI
This property has been Sold!
08/02/2016 at $53,900
4 bedroom, 2 bath home on nice large lot. Backyard is fenced, storage shed for lawn toys. Kitchen/dining area with sliders to deck. Nice 3 season porch for extra square footage. Buyer to provide approval letter from Caliber Home Loans. With offer contact Aleks Stefanovski 586-909-1235. Proof of funds letter for ALL CASH OFFERS.
Property Information
- Status
- Sold
- Address
- 1963 Hudson Street
- City
- Muskegon
- Zip
- 49441
- County
- Muskegon
- Township
- Muskegon City
- Foreclosure
- Total Finished SqFt
- 1,040
- Above Grade SqFt
- 1,040
- Garage Desc.
- 2, Attached
- Water
- Public
- Sewer
- Public
- Year Built
- 1948
- Taxes
- $1,208
Rooms and Land
- MasterBedroom
- 1st Floor
- Bedroom2
- 1st Floor
- Bedroom3
- 2nd Floor
- Bedroom4
- 2nd Floor
- Dining
- 1st Floor
- Kitchen
- 1st Floor
- Living
- 1st Floor
- Basement
- Full Basement, Walk Out
- Cooling
- Central
- Heating
- Forced Air, Natural Gas
- Acreage
- 0.22
- Lot Dimensions
- 80 x 117
- Exterior Materials
- Wood
- Exterior Features
- Deck(s), Fenced Back
Listing Video for 1963 Hudson Street, Muskegon MI 49441
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Listing Courtesy of Coldwell Banker Woodland Schmidt Muskegon
Office Address 131 Seaway Dr
Listing Agent Anne Valentine
Listings last updated: . Some properties that appear for sale on this web site may subsequently have been sold and may no longer be available.
Our Michigan real estate agents can answer all of your questions about 1963 Hudson Street, Muskegon MI 49441. Real Estate One, Max Broock Realtors, and J&J Realtors are part of the Real Estate One Family of Companies and dominate the Muskegon, Michigan real estate market. To sell or buy a home in Muskegon, Michigan, contact our real estate agents as we know the Muskegon, Michigan real estate market better than anyone with over 100 years of experience in Muskegon, Michigan real estate for sale.
The data relating to real estate for sale on this web site appears in part from the IDX programs of our Multiple Listing Services. Real Estate listings held by brokerage firms other than Real Estate One includes the name and address of the listing broker where available.
IDX information is provided exclusively for consumers personal, non-commercial use and may not be used for any purpose other than to identify prospective properties consumers may be interested in purchasing.
All information deemed materially reliable but not guaranteed. Interested parties are encouraged to verify all information. Copyright© 2017 MichRIC LLC, All rights reserved.

subset: commoncrawl


text:
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With First Tutors: Music you can find a Westhill composition tutor. Composition tuition are an excellent way to start an entertaining new hobby, pick up a new talent and attain confidence with composition. Try our service to search for Westhill composition teachers and choose which best matches your needs so you can enjoy your composition tuition in Westhill straight away!
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text:
Quake's economic effects likely to be short term
The massive earthquake that shook Japan on Friday is expected to at least temporarily set back an economy that has faltered lately and is struggling to recover from years of stagnation.
Ultimately, economists say, efforts to rebuild homes, businesses and highways in Japan will at least offset any dampening effects and possibly even spark economic growth.
The most tangible effect of the disaster on the U.S. was a drop in oil prices. In New York, crude fell $1.54 a barrel to $101.16, reversing a recent run-up fueled by turmoil in the Middle East.
Japan is the world's third-largest oil importer. Reuters reported that six refineries that account for 31% of Japan's output shut down after the quake and a fire broke out at one of the facilities.
Oil prices fell largely because the closures could limit the country's ability to purchase oil. In addition, economic growth could be crimped in the short term, says oil analyst Phil Flynn of PFGBest Research.
The Japanese yen rose against the dollar Friday. Win Thin, an analyst at Brown Bros. Harriman, attributed the rise to insurance companies converting foreign investments to the Japanese currency in advance of spending for the rebuilding.
The quake in northeastern Japan, which includes the city of Sendai, is in a relatively rural area, limiting the economic fallout. IHS Global Insight estimates the main area affected, the Miyagi prefecture, accounts for nearly 2% of the Japanese economy. Still, Toyota and Sony were among the manufacturers that shut down plants, most likely as a precaution.
"It's a shock, but it's a small shock, and it's not going to create a long-term effect," says David Griffith, professor of global marketing at Michigan State University.
IHS analyst Sara Johnson says the quake could cause the Japanese economy to contract slightly in the first quarter instead of growing slightly as her firm has been predicting.
Still, the quake came as the Japanese economy was slipping. It grew 4% last year as businesses replenished depleted inventories, but it slipped 1.3% in the fourth quarter.
The government's reconstruction should boost growth, but it will add to Japan's public debt, the world's largest at 200% of gross domestic product. A quake in Niigata in October 2004 shrank the economy 0.4% that quarter, but then it grew 1% the next two quarters, Capital Economics says.
For the U.S., the rebuilding will likely prompt the Japanese government to sell some of its holdings in U.S. Treasuries to finance the reconstruction, says John Lonski, chief economist of Moody's Capital Markets Group. Japan's 10% share makes it the second-largest holder of Treasuries after China. Its sales could push up Treasury yields and, in turn, borrowing costs for U.S. consumers.
Posted | Updated


text:
Okay, I noticed a large influx of 3DS and Skyward Sword videos (from Comic-con) in my youtube sub box, so I've decided to put them all in one, neat, organized blog.
The Legend of Zelda: Skyward Sword
This is the Comic-con trailer, unfortunately it's pretty much the same as the E3 trailer, except, now with Gorons.
This video shows Rich/Link battling a skeleton enemy that's been seen multiple times in other trailers and gameplay clips.
This shows some gameplay against Ghirahim. Actually, I haven't watched this one yet, because I don't want to spoil anything :p
Super Mario (3DS)
This gameplay vid. shows off show awesome Battleship gameplay, reminiscent of Super Mario Bros. 3
This second video shows show Tanooki suit gameplay along with the signature end-of-the-level pole.
Mario Kart (3DS)
This clip shows show footage of the Donkey Kong Country Returns themed level that Retro Studios worked on.
This clip shows off multiple levels and the voice acting - much of which is by the original voice actors!
Metal Gear Solid: Snake Eater 3D
The title may say Mario Kart, but the gameplay is actually from MGS3 3D, which has gotten the youtube video a ratio of 3 'dislikes' for every one 'like'.
So, are you as excited as I am? or not? Tell me in the comments!
Want more? Check out my second Comic-Con blog for more footage, including Kirby: Return to Dream Land and Kid Icarus Uprising.


text:
First copy text 1 (Here selected visual lines (V) and then yanked (y))
Then select text 2 visually (Here again visual lines (V))
While still in visual mode, press 'gd' to perform diff. Green marks identical text. Red marks differences.
The diff can also be done the the other way. Here we see blue marks removed text:


text:
The Maricopa Community College's Department of Public Safety has established a mission statement, vision, and motto to be consistent with and supportive of the District's vision, its noble educational mission, and its institutional values.
If you have concerns or need more information about security and safety at the Maricopa County Community College District, please contact the Chief of Police at (480) 731-8436.
Campus Public Safety Mission
The mission of the Public Safety Department at the Maricopa Community Colleges is to provide a safe and accessible learning environment, assistance and security for people on campus, protection of District and personal property, traffic control, visitor assistance, operation of a lost and found function, loss prevention/reporting, identification of safety hazards, and training/orientation of employees and students.
Public Safety Departments at Maricopa Community Colleges intend to...
- assist students in learning to resolve conflicts in a reasonable manner
- teaching them that they must take responsibility for their actions
- make students involved in their communities by reporting crimes and any suspicious activity to Public Safety
- provide a safe and secure learning environment


text: I am in the market for a used truck, and was wondering whether or not I should really concentrate on looking for at least a 3/4 or 1 ton truck. 1/2 ton are a lot easier to find, and don't cost as much, but I don't really know what they are capable of. I will probably be looking at either a Ford or Chevy. I would like to use the truck that I buy for mainly hauling my trailer and mowers, but am also considering getting into laying mulch. How many yards could I pull (with a 350 or 351) in say, a 5 X 16 dump trailer. Also, I want to add plowing as a service. I have had people tell me that it would be too much for a half-ton, and then I have had people tell me that it would be fine. Any advice?


text:
Check Out Our New Laser Cut Stencils
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text: Samuel - Unregistered November 08, 2010 12:57 AM Jense is a good guy--im more of a Lewis fan but, you know he showed his garters this year, taking two early wins. They both can learn from each other and they have a good relationship(as of now) so the team is unified and itll be strong going forward. 4 agree 0 disagree


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Customer Testimonials:
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Moms' Night Out - Combo DVD - Blu-ray
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Updated: 8/10/2016 11:27am PDT
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All Allyson and her friends want is a peaceful, grown-up evening of dinner and conversation... a long-needed moms' night out. But in order to enjoy high heels, adult conversation and food not served in a bag, they need their husbands to watch the kids for three hours—what could go wrong? MOMS' NIGHT OUT is an endearing true-to-life family comedy that celebrates the beautiful mess called parenting.
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Sarah P: I saw it at the theater and loved it!! Sep 2, 2014
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Read More»
Condition: Standard
All Product Details
Clear directions and concise descriptions lead to all of Maui's well-known attractions, as well as to hidden discoveries that Trailblazer readers have come to expect.
137 different hikes and strolls to tropical rain forests and remote valleys, coastal bluffs and lava caves, Haleakala crater and the Hana Highway, cascading waterfalls, beaches, ridgetops, towns, whale-watching perches, historic sites, and archeological ruins.
Among the 44 snorkeling spots are hike-to coves and the secret places that tour boats go.
Kayakers can pick from about 20 put-ins.
Surfers can select from 38 beaches and decide whether to boogie, board, or body surf. Onlookers will find the best places to watch the surfers, windsurfers and kite-boarders ride the big ones.
The text is complimented by 10 maps and 240 photographs. Driving tours nine of them take readers to all the attractions, natural wonders, and historic sites.
A Resource Links section provides numbers for free visitor information and recreational outfitters, as well as hand-picked accommodations and local restaurants to suit every budget and taste bud.
A Best Of section lets you pick the right activity to suit your mood and the day. Appendices include free hula shows, farmer's markets, what to pack, climate, history, fauna, and a Hawaiian glossary.
This new and completely revised third edition includes a Trailblazer Kids section for adventuring families and 4 pages of color photos.


text:
Transformers Dark Of The Moon
July 4, 2011 by Post Team
Transformers Dark Of The Moon, Transformers: Dark Side of the Moon (3-D) 3 stars Shia LeBeouf, Frances McDormand, John Malkovich, Patrick Dempsey Director Michael Bay you like it if you like this: Transformers 1 and 2, Iron Man 1 & 2, Armageddon, Pearl Harbor, Wild Wild West could Dark of the Moon is the best movie Transformers yet? Could it be possible to reverse the dramatic full of Michael Bay-edge metal that was Revenge of the Fallen, with its vision of transforming gold testicles? Could win back the hearts have 20 and 30-something loved toys that Hasbro in 1980 to meet today's rogues raised on a diet of superhero blockbusters?
I thought that after the spectacular opening sequence, if nothing else, are the best part of the Bay of mega-franchise.
Lights back on the space race by John F. Kennedy promised a man on the moon. Learn – in a twist nice conspiracy theory – which is due to a transformer called Sentinel Prime has landed on the moon, and during the famous 1969 moon landing, the astronauts will carry out a top secret mission, while the side dark moon.
These sequences are Transformers: Dark of the Moon's sense of history, humanity-into-play and – dare I say – such a lack of seriousness of the first two films. But like the Pink Floyd album of almost the same name, which is where things get a little hazy.
And our place in the war between the Autobots (candy) and Decepticons (bad) takes shape, uber-director Bay (Pearl Harbor, Armageddon), changes in the hyper-computer fast enough to overlook argument giant leaps sized robot and the script more incomprehensible, since, in addition, the latest Transformers movie (Ed Wood would be proud). Something about an "ark" that holds the key to the salvation of the Autobots.
So, before you can blink, being dazzled by one of the best models in one of the worst blockbusters.
Look! Frances McDormand is a government drone that does not like being called "Maam". Duck! John Malkovich is a control freak and corporate giving Sam Witwicky (Shia LaBeouf), his first job. Hey! It's Patrick "McDreamy" Dempsey as the head play boy Slinky Sam's new girlfriend (model Rosie Huntington-Whiteley).
Wow, this new film is actually trying to impress – there's even the real Buzz Aldrin. Ken Jeong and Alan Tudyk comic add nice touches – and they all offer to act deliberately ham is a cry of welcome. I guess I know who are in a Transformers movie.
Then come the regular stars Josh Duhamel, Tyrese Gibson, John Turturro and LaBeouf, who can be charming when he's not ranting about the ark Megatron to Cybertron close enough to Earth to enslave the planet.
And that is where Transformers 3 is transformed back into the same kind of big, loud, brash show, giddy nonsense that made the second movie so horrible.
The U.S. military are more involved than ever and, although his answer to everything seems to be "blown up"; there is an impressive scene, where the wing-suit in the training gliders weave through a maze of skyscrapers city?? (For no apparent reason, only looks good).
There is another "wow" moment that Sam flies through the air like Bumblebee transforms – in super slow motion – the car at full speed the robot to new car.
But above all, of the moon is all giant robots at war, and that's more metal crunching Bay CG-edge you can throw a plastic action figure of the cases.
There is nothing too offensive about it – no gold balls, not Megan Fox – and it looks and sounds impressive 3-D. But at 154 minutes, four minutes longer than the second half and still 30 minutes longer.
Non-stop action that leads to the point that the final action-packed chaos feels like metal. By then, I do not care who lives or dies, or if the human race is enslaved.
However, the austere opening sequences and the honor roll of the actors hamming it up of the moon makes the second best of the trilogy.
That lack of control, wit and charm of the first, but avoid the gag reflex that followed the second film abhorrent.
It's fun in an empty classroom in shape; with its effervescent energy drink will be as flat.
Please feel free to send if you have any questions regarding this post , you can contact on
Disclaimer: The views expressed on this site are that of the authors and not necessarily that of U.S.S.POST.


text:
Photo by DavidRosario.com
MIAMI (CBS4) ― Miami Heat star Dwyane Wade took a pay-cut to make it happen and late Monday afternoon; veteran power forward Udonis Haslem announced he's coming back to the Heat. The 7-year veteran confirmed his decision in a text message to the South Florida Sun-Sentinel.
"Turned down full mid-level from Dallas and Denver. See U next season," Miami Heat's #40 Haslem texted.
Haslem was rumored to be a prime target of the Mavericks, Nuggets, Jazz, and Knicks along with other teams. He could have signed for $34 million over five years based on his years in the league.
Haslem will sign a four-year deal with the Heat, though the Sun-Sentinel said contract details were still being worked out late Monday afternoon.
Haslem repeatedly said he wanted to remain with the Heat. His presence, along with the new core of Wade, Chris Bosh, and LeBron James, give the Heat a very powerful starting core to work with.
Haslem can knock down jumpers and is a solid rebounder; both things the Heat need, especially coming off the bench. The Heat are also expected to re-sign Joel Anthony, a center who is currently a restricted free agent.
The Heat also are linked to Cleveland Cavaliers free-agent center Zydrunas Ilgauskas.
Source: CBS4


text:
All is fair in politics. But to rain on a guy's golf vacation?
Mitt Romney may have broken a man-law on this one.
The 2012 presidential hopeful took a shot at President Obama's planned 17-day Hawaii vacation.
"I just think it's time to have a president whose idea of being 'hands on' doesn't mean getting a better grip on the golf club," said Romney who resents that the president is asking Congress to stay in session while he goes off for
"17 days for golf and the sun."
Couldn't this all be solved in a 72-hole match over four days with handicaps included and no pressing on the back nine?
Reid Cherner has been with USA TODAY since 1982 and written Game On! since March 2008.
He has covered everything from high schools to horse racing to the college and the pros. The only thing he likes more than his own voice is the sound of readers telling him when he's right and wrong.
Michael Hiestand has covered sports media and marketing for USA TODAY, tackling the sports biz ranging from what's behind mega-events such as the Olympics and Super Bowl to the sometimes-hidden numbers behind the sports world's bottom line.


text:
Butterick 9100; ca. 1959; Misses' Afternoon or After-Five Dress. A huge collar highlights a chic afternoon dress with side fullness in skirt. (A) Luncheon or afternoon version. (B) Dressy or bridesmaid's version with double collar, contrasting midriff.
Featured in Butterick Fashion News, November 1959
Links to reviews/blog posts Edit
Sources/Vendors Edit
Gallery Edit
[insert your photos of this pattern made up]
Wishlist Edit
[insert your username, and make sure your preferences allow for people to contact you via e-mail. Do not list your email address here!]


text:
Serpent of the People
Images of Alicia & Hannibal
by James Stanley Daugherty
About the Book
Features & Details
- Category Fine Art Photography
Standard Portrait, 8×10 in, 20×25 cm
28 Pages
- Publish Date Jan 28, 2007
- Tags BBW, Nudes, Snakes
See More
About the Creator
James Stanley Daugherty
Maui, San Francisco, Las Vegas
A Fine Art Photographer devoted to following his muse. His muse sometimes involves taking pictures of people without their clothes on.
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Overview Edit
The New York State's Committee on Open Government oversees and advises the government, public, and news media on Freedom of Information, Open Meetings, and Personal Privacy Protection Laws. The Committee offers guidance in response to phone inquiries, prepares written legal advisory opinions, and provides training to government and other interested groups. Recommendations to improve open government laws are offered in an annual report to Governor and the State Legislature.


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Obviously, you bought your RL because you understood you would be getting long-lasting quality and dependability both rolled up into one car, truck, or SUV. Acura automobile enthusiasts have come to expect a certain level of a reputation for safety when driving their RL around town. You bought your RL because its sporty handling and Japanese engineering struck a chord inside you; so why would you be okay with second rate when it comes to great quality auto parts? The vehicle in your driveway symbolizes the best in class when it comes to all the cars, trucks, and SUVs in today's marketplace.
We may take this for granted, but it wasn't until the 1950s that vehicles were equipped with reliable air conditioning. The air conditioning uses a refrigerant called R134a to fuel a thermal reaction that emits cold air, and the A/C condenser on your Acura RL acts like a radiator (it also looks like one) by cooling the refrigerant after it leaves the compressor, and then sends it to the drier and expansion valve. You can locate your Acura RL A/C condenser sitting right in front of your radiator, as that is the easiest place to soak up air while your car is driving. If your A/C condenser has sprung a leak, then you will likely notice your air conditioning blowing lukewarm air as a result of the lack of refrigerant.
Make all your future car parts purchases at carpartsdiscount.com and realize incredible savings on all your upcoming repairs. Bring your RL back to life with the best repair parts from the experts at Car Parts Discount. Don't let a A/C Condenser repair bring about the demise of your car, truck, or SUV, and don't gamble with its condition by using anything but the best replacement parts. Just choose your year from the list below to find the right Acura RL A/C Condenser for your 2012, 2011, 2010, 2009, 2008, 2007, 2006, 2005, 2004, 2003, 2002, 2001, 2000, 1999, 1998, 1997, 1996 model.


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Book your table at Bills - Cambridge
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There's a special feel to Bill's Cambridge restaurant. Tucked away on a small back street, the restaurant opens up like a Tardis once you step inside, with a bar, a library a big open dining area and a private dining space. Bill's is open for breakfast, lunch, dinner and everything in between.


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About to purchase a 4" lift (tommorrow)
Discussion in '1973-1991 K5 Blazer | Truck | Suburban' started by BowtieBlazer, Jul 19, 2002.


text: One of the biggest headline-grabbers last week was the launch of the heavily-rumoured Sony Xperia S (formerly the Nozomi), the first phone without the Sony Ericsson moniker and the company's first NXT handset. So when will it finally arrive? If Clove is right, it will be here on 5 March.That's the date when the company is said to be shipping the dual-core Android smartphone. Or at least, the week following that date. Obviously, you'll be talking SIM-free, which means laying out the money upfront. That money is quoted as £467.99.Don't worry though. If Clove has stock of the phone on 5 March, there's every likelihood that the networks will be offering contract deals around that time too. O2, Three and Orange confirmed they will be offering the phone not long after launch, with T-Mobile following and Vodafone almost certain to join the merry throng. Pricing for the networks still to be confirmed across the board.If you missed the launch, this is the first of the next generation NXT Sony Android phones, although it will only come with Android Gingerbread (2.3) at launch. An upgrade to Ice Cream Sandwich will follow in the second quarter of 2012.Other details are more impressive, with the Xperia S packing a 4.3-inch Reality Display with Mobile BRAVIA Engine and HD resolution screen (1280x720), front (720p) and back (1080p) cameras for HD recording, 1.5GHz dual-core processor, 32GB of internal flash storage, a 12-megapixel Sony Exmor R rear camera, NFC and HD sharing through DLNA and HDMI. In fact, if you hook it up to your TV, you can use BRAVIA Sync to control your phone.As soon as the pricing is confirmed, we'll let you know. Orange is promising those details 'soon', so keep an eye on the site.


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Darlene Love brings holiday show back to the Nourse
Updated 12:02 pm, Thursday, December 1, 2016
Darlene Love bristled when she first entered the studio with producer Phil Spector to record "Christmas (Baby Please Come Home)."
"I remember saying, 'This is never going to work,'" she says, calling from her home in New York. "You're going to put a song nobody knows next to 'White Christmas' and 'Frosty the Snowman'?"
For a while, it seemed as if Love, 75, was right. Released as a single from the album "A Christmas Gift for You," it was a flop upon its release on Nov. 22, 1963, the day President John F. Kennedy was assassinated. But in the years since, the song has become a seasonal classic, earning Love regular television appearances (she was one of David Letterman's favorite performers) and covers by such artists as U2, Michael Bublé and Mariah Carey.
The tune also serves as the centerpiece for Love's annual holiday show, which returns to City Arts & Lectures at the Nourse Theater on Sunday, Dec. 4.
"Isn't that a wonderful thing, to have a great, great song that you get to sing once a year?" says Love, who gained new fans from her appearance in the Oscar-winning 2013 documentary on background singers, "20 Feet From Stardom." She's also a Rock and Roll Hall of Fame inductee.
"I used to get onstage and spent 10 to 15 minutes explaining who I am," she says. "I don't have to explain anymore."
A Darlene Love Christmas: 7 p.m. Sunday, Dec. 4. $45-$75. Nourse Theater, 275 Hayes St., S.F. http://www.cityarts.net/event/a-darlene-love-christmas/


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We were given some cute little wooden chairs and I've been dying to get to work on a little makeover project, and here are the results! Its not massively ambitious, but remember it is my first go!! Let me know what you think!
and after!
here it is next to its original matching buddy! I think I want to do this one differently, maybe covering it with some french grain sack material and painting the wood red perhaps?!?
any other ideas anyone?!


text: Thank you so much for your comment, I love to hear from you. Your comment will appear in a little while when I moderate it.You might be seeing a verification thingy here, but I have verification turned off and I believe that you will be able to ignore the verification and just leave a comment anyway! Go on - try it.


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Wow, it was really hard to get all these male celebs in the same jpg together. A lot of complex scheduling, not to mention meeting all their diva-like demands. Specifically:
- Jon Hamm demanded Pantene Pro-V Conditioner for his chest hair and that his white jock perfectly match his white socks. No off white for Mr. Hamm!
- Zachary Quinto demanded a toilet to puke in. Anything for that lean and logical figure.
- Chris Hemsworth demanded to be brutally spanked. It's his thing.
- Chris Pine demanded we watch a marathon of motorcycle cop show CHiPS. His father played Sergeant Getraer. What a weird request and what a formulaic show. It's no King Lear.
- Darren Criss demanded we make sure he has a career after Glee. We couldn't guarantee that but he stuck a toy up his butt anyhow. It's a living.
- Joe Manganiello demanded tanning salon coupons and near constant head. Plus Peanut M&Ms. What a freak! Why not Charleston Chews, Joe? Why not Charleston Chews!?
- John Stamos demanded the adoration of millions of 47 year old women. And he got it.
- Ryan Reynolds demanded to be the director of Fleshjack: The Motion Picture. It will be an edgy romantic comedy starring Sandra Bullock as Pam, a lowly CEO, Ryan Reynolds as Fleshjack, a man with silicone hands, and Al Pacino as Al Pacino. Plus a special guest appearance by Shelley Long as Fleshjack's mom. You actually get to see her give birth to Fleshjack.
And that is exactly how ridiculous celebrity fakes are. But ridiculous things can still be hot, if you're in on the joke and desperately horny. And like most creative work, these say more about the creator than the subject.
Don't see the celeb you want represented? Then DIY. All you need is Photoshop (or some cheaper counterpart), a male celeb who just refused to get really naked, 37 minutes, and a dream. Just please match the skin tone of the face to the rest. Or if you can't do that so well, choose a clothed pic and Photoshop a dick sticking out of the fly. That way, people will totally believe Chris Pratt is posing on the red carpet with an exposed boner. Oh wait, it's Chris Pratt. He'd probably actually do that.
Want some actually real naked male celebs? Check out a review of Mr. Man, our highest scoring male celeb site. Star Male is right up there too. Want more fakes? Then try meeting guys on OkCupid.


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What is JotForm?
JotForm is a free online form builder which helps you create online forms without writing a single line of code. No sign-up required.
At JotForm, we want to make sure that you're getting the online form builder help that you need. Our friendly customer support team is available 24/7.
We believe that if one user has a question, there could be more users who may have the same question. This is why many of our support forum threads are public and available to be searched and viewed. If you'd like help immediately, feel free to search for a similar question, or submit your question or concern.
How to require answer to each question in a matrix without the font of the questions disappearing?Asked by nmhgmarketing on November 04, 2016 at 09:14 AM
I have an evaluation form that I am trying to implement and am using a matrix with radio buttons for survey-takers to answer.
When I require an answer to each question, once you select the first radio button the font seems to disappear. The only way I've been able to fix this is to select "no" to requiring an answer to each question. But our team does want to require this. How can it be fixed?
JotForm Support
Adding the following code to your form will change the color of highlighted text when the error is present:
.form-line-error {
color: black;
Here is how the form would look with the code added:


text:
Lightly scented promotional hand sanitizer spray that fits conveniently in your pocket or purse.
Stay in style with the Double Wall Custom Tumbler - it's the perfect item to carry your hold and cold beverages and keep them to the perfect temperature.
Made of BPA-free PETG plastic this custom promotional water bottle is both durable and a healthy choice for your logo.
A clear flexible double walled econo promotional tumbler with a straw and lid features your custom logo.
A handy little logo flashlight that you can clip anywhere.
Carry hot or cold beverages in style with these large custom tumblers that are double wall insulated to keep drinks at the perfect temperature.
8-ounce non-spill logo drinkware for babies with 2 handles is perfect for new and expecting parents.
A successful promotional campaign is right around the corner with education promotional items from ePromos. We have plenty of items for multiple areas of education such as school, art, dance, online training and automotive. Whether you're looking for specialized products for each field or looking for promotional pens, or promotional stress balls that can be used for any type of field, ePromos has what you are looking for.


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Belvedere Locksmiths in Belvedere
We are a Belvedere Locksmiths based company, providing fast, professional & reliable locksmith services. We also provide a 24 Hour emergency Locksmith services at competitive prices. Call us on Tel: 020 8819 8618.
Emergency Locksmith Belvedere
When you are locked out, lost your keys, or simply your lock is not working, please contact us anytime and we will be with you in less than 30 minutes of your call.
Locks Change
Our locksmiths are available to change your locks in your home and office anytime. We also install British Standard Locks for extra security. Locks include Yale, Chubb, 5 Lever Deadlocks, Euro Locks, Window Locks and bolts and many more.
Please contact us for more information.
British Standard Locks
Our locksmith carries all locks, and always gives our customers the best prices for British Standard High Security Locks, which conforms to Insurance Companies requirements.
Daniel D
020 8819 8618020 8819 8618
Shortlands Close,DA17 5QU


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Richard Branson Is Launching an iPad Publication, Too, You Know
"We definitely haven't got a Rupert Murdoch–size advertising budget ... "By Mike Vilensky
"We definitely haven't got a Rupert Murdoch–size advertising budget ... "By Mike Vilensky
The Producers of HBO's Confederate Respond to the Backlash and Explain Why They Wanted to Tell This Story
Dunkirk Is a Great War Movie Marred by Christopher Nolan's Usual Tricks
Trump Is Mobilizing for War Against the Rule of Law
The Triumph of Obama's Long Game
A Conversation About Naomi Watts's Instagram, the Summer's Best Feed
All 10 Christopher Nolan Movies, Ranked
When Will The Planet Be Too Hot For Humans? Much, Much Sooner Than You Imagine.
Donald Trump Could Really Use a Sober, Senior, Reassuring Communications Director. Instead He Chose the Mooch.
You Can Tell If Someone's Rich or Poor by Looking at Their Face
Gigi and Bella Hadid's Dad Was Just Sentenced to Probation for the Most Rich-Person Crime


text:
Pet Sitter in Pittsburgh
Hello, My name is Moniffa(Moe-knee-fa), but I go by Moe. I currently do not have experience walking dogs or caring for them and I think this is the perfect way to learn. I absolutely love animals. I prefer a dog over a cat, even though I have 2 cats :) I think this would be a great way for me to make some money while building relationships with dogs and their owners as well.
Pet Sitter Details
|Email Address
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Dog Walking
Pet Sitting
Notes made here will only be seen by you. If you want to message the owner of this listing
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text:
Update at 1/7/11, 10:47 a.m. ET: Apple says the app store has already topped one million downloads in its first day.
"We're amazed at the incredible response the Mac App Store is getting," said Apple CEO Steve Jobs in a statement.
Our original post
Apple made some news at the same time as the Consumer Electronics Show, but it's not a Verizon iPhone (at least not yet) and this news was already quite expected. The company has just opened business on its Mac App Store.
The store launches with more than 1,000 free and paid apps and is available for Snow Leopard users who fetch version 10.6.6 (use Software Update for the download.) "With more than 1,000 apps, the Mac App Store is off to a great start," says Apple CEO Steve Jobs in a statement. "We think users are going to love this innovative new way to discover and buy their favorite apps."
The Store is broken up into categories (Education, Games, Graphics & Design, Lifestyle, Productivity and so on.) As with Apple's similar App Store for the iPhone and iPad, you can get a read on what's hot, staff favorites and look up charts for top paid and top free apps.
The Autodesk SketchBook Pro drawing program, Ancestry.com's Family Tree Maker and Boinx Software's animation and video production software are among the apps available at launch. And Apple is now selling iLife '11 apps iPhoto, iMovie and GarageBand individually for $14.99 each, and Pages, Keynote and Numbers (from Apple's iWorks suite) for $19.99 each. Apple's Aperture photo editing program can be had for $79.99.
Of note: when you buy an app, you can download it to any of your personal Macs, so long as they're authorized with your AppleID/iTunes account. There is no limit. I'll say more when I've had a chance to check it out the new store firsthand.
In the meantime don't hold your breath for that Verizon iPhone being announced at CES, though Verizon will be heard from later in the day.
By Ed Baig
Technology Live covers computers, electronics, and the Internet, with a focus on cutting-edge technology that has a real impact on your life. Our reporters and editors -- based in San Francisco, Los Angeles, Seattle, New York, and Washington – offer news and tips to help you make the most of your digital domain. More about the team
Sign up for Technology Live e-mail alerts


text:
If there's one thing Williams might want to think about, it's scaling back the defensive schemes into something more manageable. Ted Cottrell had his standard 3-4 defense, then on third downs he went to a 4-1-6, 4-2-5 or 3-3-5. Toward the end of his Bills career, it was a 3-3-5 with Sam Cowart, Sam Rogers and Keith Newman staying in the game to rush the passer, John Holecek and Pat Williams coming out, with an extra DB coming in – usually Donovan Greer. Occasionally, he'd put together something special, like "the Package," with Pat Williams and Ted Washington up front, which worked on Williams' Titans pretty well during last season's opener.
But Gregg Williams has all kinds of schemes and groupings. During the off-season, he even bragged about the fact that the Bills have something like 13 defensive packages. They may have all those, but they're certainly not playing them well. It might take them a season or two to do the things Williams is asking them to do. How about doing a couple formations and schemes well for the time being?
Now you hear a lot about Williams' 46-defense, but let's explain things according to positions. For most media people, referring to the "complexities of the 46 defense" without explaining what's happening on a play is just an easy way for them to say, "I don't know what the hell is happening myself."
During the four big plays that produced or helped produce Indy's first four touchdowns, the Bills were in four different modes of defense. This is not all that unusual, except for that different personnel are on the field. Suffice it to say, this causes limited continuity among players, and blended with the fact that these players haven't played that long together, it's a recipe for the problems that were seen in the RCA Dome.
On the 45-yard long-gainer featuring Peyton Manning to tight end Ken Dilger, which set up Colts' touchdown No. 1, the Bills were playing a 5-3. Aaron Schobel was right end, Shawn Price weakside tackle, Pat Williams nose tackle, Phil Hansen strongside tackle, and Erik Flowers left end. Brandon Spoon was middle linebacker, with Keith Newman and Jay Foreman both lining up on the strong side, over Dilger. Newman was inside and Foreman outside.
A 5-3? What the heck's that? Well, it's a short yardage defense. It was third and one and the Bills wanted to make sure that Edgerrin James wasn't going to run for the first down. He didn't. Instead, Manning faked the handoff to him, sucking Keith Newman in, while Dilger initially blocked Phil Hansen, then ran free like an antelope on the open prairie. Foreman covered tight end Marcus Pollard, who ran a short crossing pattern as the flanker, away from where Dilger was running, effectively taking Foreman out of the play. Manning dumped off to Dilger in the right flat and it turned into a 45-yard gain.
Interestingly, Erik Flowers was playing left end – just to the left of the two tight ends in that Colts offensive formation – and Bryce Fisher right end, which doesn't make a whole lot of sense because Flowers is not a great in-traffic type of player. That's why he was ineffective in the 3-4, playing over the right tackle when he subbed for Marcellus Wiley last season. But that's what the Bills did this time.
On the flea flicker 60-yard touchdown to Jerome Pathon, it was a first and 10 and Buffalo was in a nickel package. A nickel package! On first down! The Colts had a slot receiver and wide receiver to the right. They had their two-tight ends flanking the line. They certainly looked like they were going to run it. Of course, to make things ironic, a nickel package is designed to look for the pass first. But no one was looking for the pass in this case. Antoine Winfield bit on the fake and released Pathon, the slot receiver. He ran by Keion Carpenter and caught the ball for the score. Everybody was converging on James upon the handoff.
On the 39-yard sideline touchdown to Harrison, the Bills were playing a standard 4-3. Again, the line was jumbled though. Flowers played right end, Leif Larsen was strongside tackle, Pat Williams weakside nose tackle and Aaron Schobel was the left end. This time Manning stutter-stepped and squared his shoulders toward the sideline like he was going to throw, which caused Winfield to hesitate and Harrison to get free as the quarterback actually released the ball – another easy score.
On the 39-yard slant touchdown to Harrison in the middle, the Bills were in a 4-4-3, which is a formation, again, designed to stop the run. Schobel was the right end, Pat Williams the weakside nose tackle, Shawn Price the strongside tackle and Phil Hansen the end. Jay Foreman was weakside outside linebacker, Brandon Spoon weakside inside linebacker, Kenyatta Wright strongside inside linebacker and Newman strongside outside linebacker. Another run fake caused Winfield to stay in, which released Harrison, who ran past Carpenter, and caught the pass for another 39-yard touchdown. Strong safety Raion Hill was taken out in favor of the extra linebacker – in this case Kenyatta Wright.
So what did we learn from Sunday's game? Williams likes to use varied defensive personnel because it proves how great a coach he is. But the varied personnel means nothing if it can't play together and/or it can't identify run fakes, pump fakes and draw plays.


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May Chang (Litsea Tree)
Essential oil is extracted from fruits of the tree and have very refreshing and stimulating effects. The fragrance of may chang is both uplifting and relaxing and creates feelings of optimism and confidence. Due to its antifungal and antibacterial properties, may chang can be used to help treat acne, eczema and other skin conditions, and is a popular additive to natural insect repellants.
May Chang & Lemon Herbal Bath Oil - "Enjoy Life"
Herbal Bath Oil3.38 fl.oz.$20.00
May Chang & Lemon Body Wash - "Enjoy Life"
Body Wash6.76 fl.oz.$14.00
May Chang & Mint Aromatherapy Bubble Bath - "Enjoy Life"
Bubble Bath13.52 fl.oz.$15.00
Mini May Chang & Lemon Bath Oil - "Enjoy Life"
Herbal Bath Oil.67 fl.oz.$4.00
Mini May Chang & Lemon Body Wash - "Enjoy Life"
Body Wash2.53 fl.oz.$6.00


text:
Daventry was a member of the Coalition of Ordered Governments Air Corps King Raven maintenance crews.
Lambent InvasionEdit
- "Colonel, we can't stay here now. Are you going to evacuate? I know it's going to be tough on the mainland, but at least we can run away from these things there."
- — Daventry to Hoffman, after the Lambent attacked Vectes Naval Base
Shortly after the Third Battle of New Jacinto, Daventry joined several others outside of the infirmary building of Vectes Naval Base, waiting for Col. Victor Hoffman to come out. When he did, Daventry said that they couldn't stay on Vectes anymore, and asked if they were going to evacuate. He told Hoffman that he knew things would be tough on the mainland, but that they could at least run away from the Lambent there. Hoffman told him that they would be making a decision soon, and to just hang in there until they did.[1]
- ↑ Gears of War: Coalition's End pg 387


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Drop N' Dash
December 22-23 & 27-28, 2016
Kids stay active and out of the way, while parents shop for the holiday! Join Dudziak's this winter for two fun filled days with gymnastics, games & activities, arts & crafts, and a movie! Need more time at camp? Dudziak's offers early drop off (8:00am) and late pick up (4:00pm)! Call the Main Office at 207-286-3685 for more information!


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America's first seaside resort doesn't readily embrace change, and that's a good thing. Visitors explore the Victorian gingerbread buildings of the historic district on bicycle, carriage, and trolley bus. They sleep amid floral-print wallpaper, on four-poster beds, in homes formerly owned by bluebloods of the 19th century. They dine and drink where U.S. presidents once did. And when flocks of migrating birds arrive at the same time every spring and autumn, they put the binoculars to their eyes and see yet another thing that has enticed beach pilgrims for over two hundred years.
Do you like being alive? Don't go to these places.More


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The game will be aired live on ABC.
The Hurricanes will take on the Seminoles for the 61st time in program history. Miami owns a 31-29 series advantage against Florida State.
For the latest information on the Miami Hurricanes, follow us on Twitter and Instagram, @CanesFootball or visit our website at HurricaneSports.com.


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|Message view
|« Date » · « Thread »
|« Date » · « Thread »
|Doug MacEachern <do...@covalent.net>
|Re: .libs/httpd
|Fri, 24 Aug 2001 04:47:36 GMT
whoops, file sizes are the same, i made a change before make install again. but i still have the same problem, they are still different somehow. i had done this: make distclean cvs up buildconf configure ... && make depend && make yet somehow (note warning): (gdb) b apr_filepath_root Breakpoint 1 at 0x4003655f: file filepath.c, line 102. (gdb) c Continuing. Breakpoint 1, apr_filepath_root (rootpath=0xbffff774, inpath=0xbffff784, flags=0, p=0x825570c) at filepath.c:102 warning: Source file is more recent than executable. 102 if (**inpath == '/') (gdb) info source Current source file is filepath.c Compilation directory is /home/dougm/ap/httpd-2.0-prefork/srclib/apr/file_io/unix/ Located in /home/dougm/ap/httpd-2.0-prefork/srclib/apr/file_io/unix/filepath.c Contains 324 lines. Source language is c. Compiled with stabs debugging format. (gdb) q % ls -l /home/dougm/ap/httpd-2.0-prefork/srclib/apr/file_io/unix/filepath.c -rw-r--r-- 1 dougm dougm 11237 Aug 23 20:33 % wc -l /home/dougm/ap/httpd-2.0-prefork/srclib/apr/file_io/unix/filepath.c 324 % ls -l ~/ap/httpd-2.0-prefork/.libs/httpd -rwxr-xr-x 1 dougm dougm 4318551 Aug 23 20:51 if i edit/save filepath.c and make, everything relinks and i still have the same problem. after 'make install', the installed binary works as expected, as does .libs/httpd. but, change a file, run 'make', appears to relink .libs/httpd, yet it behaves like the old version. very weird.
|View raw message


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''Being surrounded by underpaid, overworked and dissatisfied advertising people, and being one of them myself at that time, this song was composed years back. Released it with 4 Tune Kookies now, after meeting a new generation of more underpaid, overworked, and dissatisfied advertising people. In our time, we just cribbed. Nowadays people are actually falling sick. It's that serious. It's no coincedence that the director of the video, Naren Multani, and the actor, Manoj Shetty, are people with years of advertising work behind them.''
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text:
Get Loud, Anywhere
The Legendary Portable Pignose 7-100 amplifier is perfect for any traveling musician, student learning an instrument, or on the road rock star. The pignose is considered the ultimate portable bluse guitar amp… and being that it's battery powered, lightweight and completely portable, the Pignose can be used on the street or in a practice studio giving you a great sound any time, any where.
This 5 lbs of anytime jamming portable blue guitar amp is still used by some of the biggest names in music as a pre-amp for studio recordings. It's only $75!


text:
Economic achievement in the golden age of Islam
Discovery of Chinese papermaking process.
It was faster and easier than making parchment or papyrus, so writings were more accessible. It spread quick through the empire, from government offices to society in general. Shops started selling paper and books and business was done using credit equivalent to modern day checks.
Intellectual achievement in the golden age of Islam
House of Wisdom
Founded by Al-Ma'mun in Baghdad. It attracted scholars from surrounding continents where they translated a great amount of literature and knowledge to Arabic. They translated Aristotle, Euclid, and important thinkers of ancient Persia, Greece, and Rome. They sought to understand these and organize into Islamic teachings
Political achievement in the golden age of Islam
Islamic law
More groups came under Islamic control as the Abbasid armies spread borders and Islamic law was universal through the empire. The knowledge of different cultures blended in the empire, trade increased, and goods and information exchanged over long distance.
Social achievement in the golden age of Islam
Spread of Arabic
Arabic was the official language of the Muslim Empire. More people learned it as the empire expanded and it made it easier to communicate
Land of Great Achievement
The Islamic Empire in the Golden Age
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Economic achievement in the golden age of Islam
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text:
But the former Australian Test fast bowler is saddened at the crisis engulfing the sport in the troubled African nation.
As debate rages over whether Australia should go ahead with their tour, the moral question of playing in Zimbabwe, under the regime of Robert Mugabe, has been complicated by the inept performances of the country's drastically-weakened team against Sri Lanka.
With most of Zimbabwe's best players not available for selection as they continue to support recently-deposed captain Heath Streak, the team has been left in tatters and was bowled out for a world record 35 in the third One-day match against Sri Lanka on the weekend.
Rackemann, who coached Zimbabwe from December 1999 to October 2001, said a visit by the world's top team would provide a shot in the arm and help stave off fears cricket could be doomed in Zimbabwe.
"International tours are vitally important for Zimbabwe, obviously to fund the game through TV rights, but also for the juniors and the badly-needed development of the game for the sake of the kids," Rackemann said.
"It's disappointing to see there's players out and they're having those difficulties, so obviously I'm hoping to read some good news."
Australian cricket officials are yet to receive any new concerns from players preparing to depart on May 13 following the decision of leg-spinner Stuart MacGill not to make himself available on moral grounds.
Cricket Australia and the Australian Cricketers Association (ACA) hold grave concerns over the situation in Zimbabwe and ACA chief executive Tim May said it had the potential to cheapen Test cricket.
"There's no doubt about it, it devalues international cricket," May said on Tuesday. "We won't be surprised at what may happen with a substandard side."
Australia can only pull out of the tour through Government intervention or because of security fears.
Former Zimbabwe all-rounder Eddo Brandes on Tuesday joined the chorus urging Australia not to tour.
Brandes, now coaching on Queensland's Sunshine Coast after playing 10 Tests for Zimbabwe in the 1990s, said unless his former teammates returned to the side the coming matches would serve no benefit to either side.
"The players won't be enjoying it, that's for sure," Brandes said.
"If I was an Australian player I wouldn't want to be playing against substandard opposition and I'd want to have the trip postponed.
"If it can't be sorted out it's the history of the game that is being affected and even Sri Lanka would not have enjoyed bowling a side out for 35 and you can bet the side being annihilated wouldn't have enjoyed it."
Brandes and his family settled in Australia partly because of the troubles in their former home under the Mugabe regime.
"I have three daughters aged nine, seven and five and the future for them looked increasing difficult," Brandes said.