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St. Margaret's Centre definitely has something for everyone. Check out our Program Guide, website, and Fitness Brochure to find that special program just for you.
Leblanc TaeKwon-Do
LeBlanc Taekwon-Do is Tantallon's largest Family Martial Art Club. Come in and try a free week and get started on your journey to Black Belt. Taekwon-Do builds strength, flexibility and confidence. We are open all year and anyone can join in at anytime. Children under 10 can be a part of our Lil' Ninja Program.
Visit our website at www.leblanctkd.com or call 789-0032 for more details.
Indoor Cycling (Spin) Classes
Do you want:
- A great cardio workout in a supportive environment?
- An opportunity to increase your strength and endurance?
- Cross-training for running and other sports?
- Cycling-specific workouts and training?
You will find all that and more here. These aren't your traditional 'spin' classes. The instructor, Chris Nielsen, is an internationally recognized educator of cycling instructors. She works hard to keep things fresh with lots of different types of music and a variety of workouts throughout the week and year. She is committed to ensuring that you will feel safe and can work at your own level. Our small class sizes ensure that you will receive a lot of individualized coaching. Bring water and towel. Please arrive ten minutes early for your first class, in order to be set up on a bike. Be prepared to get hooked on the feeling.
Classes are now finished for 2016-17 season. Watch our website for schedule and start up in the Fall.
Tuesdays & Thursdays, 6:15 - 7:15 AM
Tuesdays & Thursdays, 8:45 - 9:45 AM
Tuesdays, 10:00 - 10:30 AM
Tuesdays, 7:00 - 7:30 PM
Tuesdays & Thursdays, 7:45 - 8:45 PM
Saturdays, 9:00 - 10:00 AM
Saturdays, 10:15 - 10:45 AM
1/2 Hour class, $8.00
1 Hour class, $12.00
Discounted Punch Passes for 10 one hour classes are available for $100.00
SUMMER SCHEDULE: July 5th - August 31st.
Mondays & Wednesdays, 6:00 - 9:00 PM Novice/Recreational
Thursdays, 6:00 - 9:00 PM Intermediate/Advanced
Sundays, 5:00 - 7:00 PM Novice/Recreational
Sundays, 7:00 - 9:00 PM Intermediate/Advanced
If you haven't tried it, you don't know what you're missing. September - June. Join the gang for Pickle Ball. Drop in fee of $3.50 and all ages are welcome. 10 visit punch pass available for $30.00. No matter if you're a seasoned pro at it or a beginner, come along and see for yourself how much fun it is. Try it, you'll like it.
September - June schedule:
Beginner -- Tuesdays and Thursdays, 3:30 - 6:00 PM
Competitive -- Fridays, 1:00 - 4:00 PM
Intermediate -- Tuesdays and Thursdays, 9:00 AM - Noon
Open -- Mondays, 9:00 AM - Noon
Recreational -- Tuesdays and Thursdays, 12:30 - 3:30 PM, and Fridays, 9:00 AM - Noon
Monday afternoons, 12:00 - 3:00 PM. Wednesday mornings, 9:00 AM - noon. Meet your friends at the gym, and catch up on all the news while getting some exercise. $2.00 fee applies.


text: Toy Biz made a line of Muppet Babies plush in 1997. Baby KermitBaby GonzoBaby PiggyAdd a photo to this gallerySee also Muppet Treasure Island plush Retrieved from "http://muppet.wikia.com/wiki/Muppet_Babies_plush_(Toy_Biz)?oldid=930796" Ad blocker interference detected! Wikia is a free-to-use site that makes money from advertising. We have a modified experience for viewers using ad blockers Wikia is not accessible if you've made further modifications. Remove the custom ad blocker rule(s) and the page will load as expected. Categories: Muppet Babies Plush

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text:
Mexico Reinstates Teacher Evaluations After Elections
The Mexican government has reinstated its plan to implement evaluations for public school teachers in the country, after suspending it last month amid protests by teachers.
The Mexican government has reinstated its plan to implement evaluations for public school teachers in the country, after suspending it last month amid protests by teachers.
Members of radical wing of Mexico's national teachers' union went on an indefinite strike this week in an attempt block President Enrique Peña Nieto's education-overhaul package, according to news reports.
Uncertainty over a salary proposal, a state supreme court decision on school funding, and a class-size referendum all propelled the walkout.
A judge's ruling last summer to overturn teacher-protection statutes was thinly argued and misread state constitutional law, they contend.
Fearing its members will be penalized, the union wants to eliminate the tax on high-cost health-care plans
After nearly a year of negotiations and the threat of a strike, the Los Angeles school district has reached a tentative agreement with the teachers' union, settling on a deal that would increase teacher salaries by 10 percent.
A first look at the 1.6 million-member union's take on the Senate's draft bill to rewrite the No Child Left Behind Act.
The 3 million-member union wants a revised law to push harder on exposing and rectifying inequities in funding and teacher quality.
The tentative framework would tighten tenure and dismissal processes and require the state education department to develop a new teacher-evaluation system.
About 290 teachers have been removed from a costly pool of teachers who are on the city payroll but don't have permanent jobs.


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Vascular smooth muscle cell (SMC) migration is a hallmark of intimal hyperplasia (IH), the progression of which is affected by hemodynamic conditions at the diseased site. The realization that SMCs are exposed to blood flow in both denuded vessels (direct blood flow) and intact vessels (interstitial blood flow) motivated this study of the effects of fluid flow shear stress (SS) on SMC migration. Rat aortic SMCs were seeded onto Matrigel-coated cell culture inserts, and their migratory activity toward PDGF-BB when exposed to SS in a rotating disk apparatus was quantified. Four hours of either 10 or 20 dyn/cm2 SS significantly inhibited SMC migration to the bottom side of the insert. This inhibition was associated with downregulation of SMC matrix metalloproteinase (MMP)-2 activation. Four hours of 10 dyn/cm2 SS also drastically increased SMC production of NO. A NO synthase inhibitor (NG-nitro-l-arginine methyl ester; 100 μM) abolished the shear-induced increase in SMC NO production as well as the inhibition of migration and MMP-2 activity. A NO donor (S-nitroso-N-acetyl-penicillamine; 500 μM) suppressed SMC migration via the reduction of both total and active MMP-2 levels. Addition of 10 μM MMP-2 inhibitor I to inserts significantly reduced SMC migration. Western blots showed no effect of 4 h of 20 dyn/cm2 SS on SMC production of PDGF-AA, another chemical known to suppress SMC migration. Thus it appears that SS acts to suppress SMC migration by upregulating the cellular production of NO, which in turn inhibits MMP-2 activity.
- vascular smooth muscle
- intimal hyperplasia
- interstitial flow
the development of vascular pathologies such as intimal hyperplasia (IH) and atherosclerosis has been attributed to, among other things, hemodynamic conditions at the diseased site. In particular, it has been well established that the progression of these diseases is inhibited in areas exposed to elevated levels of blood flow shear stress (SS) and exacerbated in low-SS areas (2, 11, 19, 20, 24). Abnormal vascular smooth muscle cell (SMC) migration and proliferation are recognized as hallmarks of IH development. It has been shown in balloon catheter injury animal models (19) that intimal thickening in the absence of the inner endothelial cell (EC) lining is accelerated in low-flow (SS) environments, suggesting that SMCs respond directly to changes in blood flow. In that study (19), the intimal thickening response to decreased flow levels was a result of increased SMC migration compared with high-flow conditions.
In vitro studies of the effects of SS on vascular cells have focused primarily on ECs (see, e.g., Refs. 1, 6, 8, 16, 36) because, under normal physiological conditions, they are directly exposed to the SS of flowing blood, having a magnitude on the order of 10 dyn/cm2 (26). It had long been thought that the underlying SMCs were shielded from blood flow and were only subjected to SS when the EC layer was denuded, as in balloon angioplasty, directly exposing SMCs to blood flow SS. Recent modeling studies have shown, however, that even in the case of an intact endothelium, SMCs are subjected to SS through exposure to interstitial flow driven by the transmural pressure gradient (39, 43). Although the magnitude of the superficial velocity of this flow is low (order of 10−5-10−6 cm/s, roughly 4–8 orders of magnitude lower than average velocities throughout the circulation), the interstitial spaces are very small (on the order of nanometers), resulting in SS levels estimated to be the same order as those experienced by ECs due to direct blood flow.
Although the effects of SS on SMCs are not as well characterized in vitro as those on ECs, there have been several studies suggesting that elevated SS levels inhibit SMC proliferation rates (30, 38, 42), agreeing well with in vivo work (2, 11, 19, 20, 24). One study hypothesized that SS inhibition of SMC proliferation is due to altered SMC production of TGF-β1 in response to SS, an autocrine control mechanism (42). The direct effects of SS on SMC migration are not as well studied. In the only investigation we are aware of that examined the effects of SS on SMC migration, Palumbo et al. (29) used a cone-plate apparatus to subject bovine aortic SMCs to SS in culture. After SS exposure, the cells were trypsinized and incorporated in a variation of the Boyden chamber assay to determine their migratory activity. Fifteen hours of 12 dyn/cm2 SS inhibited SMC migration by decreasing matrix metalloproteinase (MMP)-2 activator, membrane-type MMP (MT-MMP; an activator of latent MMP-2 on the cell surface), and PDGF receptor-β levels. Although this study provided valuable insight into the role of SS on SMC migration, the cells were not sheared directly while migrating, and they were subjected to harsh, nonphysiological treatment (trypsinization) during the experiment. Therefore, it is important to conduct quantifiable experiments under conditions in which SMCs are subjected to SS directly while migrating through a representative extracellular matrix.
The MMPs and PDGF, examined by Palumbo and colleagues (29), are generally considered to play a major role in SMC migration. MMPs are a family of endopeptidases with the ability to degrade extracellular matrix molecules (12). Zempo et al. (45) highlighted the importance of MMPs in SMC migration by using a chemical MMP inhibitor that effectively suppressed SMC migration through a representative extracellular matrix (Matrigel) in vitro. In particular, it has been shown that MMP-2, a gelatinase that is secreted in a proenzyme form and activated on the cell surface, is necessary for rat aortic SMC migration through Matrigel in vitro (33), and Kenagy et al. (18) demonstrated that MMP-2 plays a role in primate SMC migration through native matrix.
PDGF has long been recognized as a chemoattractant for SMCs, but the various PDGF isoforms appear to have different mitogenic properties. Whereas PDGF-BB is accepted to be chemoattractive to SMCs, PDGF-AA has been shown (22) to have very little, if any, chemoattractive activity for SMCs by itself and, in fact, appears to inhibit SMC migration when used in combination with PDGF-BB, compared with PDGF-BB used alone (21). The production of PDGF by SMCs is affected by alterations in flow. Both elevated (41) and reduced (23) flow (SS) in vivo have been associated with increased SMC PDGF-AA mRNA expression. Sterpetti et al. (37) reported increased SMC synthesis of PDGF in response to increased flow levels in vitro but did not specify which isoform was being produced. However, Barrett and Benditt (3) reported that SMCs produce PDGF-AA primarily, and other work (25) has shown that only a small percentage (1.6%) of SMCs cultured from normal medial tissue express PDGF-BB mRNA. So it is likely that the PDGF production reported by Sterpetti and colleagues was PDGF-AA. It is plausible, therefore, that SS acts to increase SMC PDGF-AA production, which acts to inhibit their migration in an autocrine fashion.
Another diffusible substance known to affect SMC function is NO. SMCs produce both NO (31) and inducible NO synthase (NOS) (14) in response to elevated SS levels, and other studies have demonstrated that NO inhibits SMC migration (13, 34). The intracellular concentration of NO in a vessel has also been linked to the activity of MMPs in that vessel (40). Therefore, it is plausible that, in response to SS, SMCs produce NO, which in turn affects their migratory response.
In the present study, we plated rat aortic SMCs on Matrigel-coated cell culture inserts and observed their direct migratory response to SS applied to their surface in a rotating disk apparatus. We quantified migration levels and used biological activators and inhibitors to evaluate our working hypotheses that 1) SS acts to inhibit SMC migration, 2) this inhibition is due to increased SMC production of NO that acts to suppress MMP-2 activity, and 3) this inhibition is due to the increased SMC production of PDGF-AA, which reduces SMC migratory activity.
SMC isolation and culture.
Rat aortic SMCs, positively characterized by both their "hill and valley" growth pattern and staining for smooth muscle-specific α-actin (monoclonal anti-α-smooth muscle actin clone 1A4; Sigma), were obtained from male Sprague-Dawley rats weighing ∼150 g via a primary culture procedure described elsewhere (44). This procedure was approved by the Pennsylvania State University Institutional Animal Care and Use Committee. Cells were then expanded in 75-cm2 culture flasks housed in a 37°C, 5% CO2-95% air incubator with DMEM-F-12 (Sigma) supplemented with 10% FBS (HyClone) and 100 U/ml penicillin and 100 μg/ml streptomycin (1% P/S; Sigma) serving as the culture medium. Medium was changed every 2 days, and cells were split on reaching confluence (4–5 days) with a 0.07% trypsin-EDTA solution (Sigma). Passages 3–9 were used in experiments.
Migration insert preparation.
Cell culture inserts (8.0-μm pore size, Becton Dickinson) housed in six-well companion plates (Becton Dickinson) were coated with 750 μl of growth factor-reduced Matrigel matrix (GFR Matrigel; BD Biosciences) diluted in Dulbecco's PBS (Mediatech) to a concentration of 476 μg/ml and then placed in a 37°C, 5% CO2-95% air incubator for 3 h to provide sufficient time for the Matrigel proteins to gel. Plates containing inserts were then placed into a hood previously sterilized by exposure to ultraviolet light and allowed to air dry for 48 h. Once dry, plates were stored at 4–8°C until needed for experimentation. This protocol reproducibly yielded a 5- to 7-μm GFR Matrigel coating thickness near the center of the insert as determined by scanning electron microscopy. Uniformity of the Matrigel coating was assessed via the use of a Coomassie Brilliant Blue R250 (Sigma) staining procedure.
Migration experiments.
A modification of the traditional Boyden chamber assay, used by others to investigate SMC migration in vitro (see, e.g., Refs. 21, 33), was used in these experiments. Inserts that had been coated as described above were first rehydrated in PBS for 1 h in a 37°C, 5% CO2-95% air incubator. PDGF-BB (Sigma) was then diluted to a final concentration of 100 ng/ml (32) in DMEM-F-12 + 1% P/S, and 2.5 ml of this solution was placed in the well of the companion plate to serve as the chemoattractant. SMCs were then trypsinized from culture flasks, centrifuged, and resuspended in DMEM-F-12 + 10% FBS and 1% P/S at a concentration of 250,000 cells/ml. This concentration provided a confluent SMC monolayer on the GFR Matrigel coating. The PBS added previously was removed, and 2 ml of this cell solution was added to the insert. Depending on the experiment, the inserts were then incubated for 6–9 h. It is of note that cells seeded on GFR Matrigel-coated inserts in serum-free medium in the presence of 100 ng/ml PDGF-BB in the well of the companion plate did not migrate on timescales up to 24 h. In experiments conducted to determine the role of NO produced by SMCs on their migration and MMP-2 activation, a NOS inhibitor [NG-nitro-l-arginine methyl ester (l-NAME), 100 μM; Sigma] was added to inserts 1 h after initial cell seeding. Inserts were then returned to the incubator and later exposed to SS as described below.
After incubation, inserts to be used in SS experiments were moved to a hood containing a rotating disk apparatus (see Ref. 36 for a thorough description) and were subjected to 1–20 dyn/cm2 SS for 1–4 h. This SS was calculated with the equation τ = μωr/h where τ is the SS, μ is the fluid viscosity (1.2 cP), ω is the rotational velocity of the disk (23.9, 239, or 478 rpm for 1, 10, or 20 dyn/cm2 SS, respectively), r is the disk radius (11.12 mm), and h is the gap distance between the disk and the SMC surface (251 μm). All SS magnitudes presented here are maximum values, and magnitudes ranged from 0 dyn/cm2 (center of insert) to a maximum at the edge of the insert, with the average value being two-thirds of the maximum. Inserts were maintained at 37°C in 5% CO2-95% air during the entire SS exposure. There was no significant change in cell morphology due to exposure to SS. Total experimental time from cell seeding on the insert to completion of SS exposure was 10–11 h. For example, if an insert was incubated first for 6 h, it was then subjected to SS for 4 h. This time course was determined to be optimal through an initial set of experiments in which it was observed that after 6–7 h cells had attached to the GFR Matrigel coating firmly enough to withstand shear exposure but none had yet migrated to the underside of the insert (data not shown). Inserts used as controls were also housed in the experimental hood with a rotating disk in place, but the disk remained stationary throughout the duration of the experiment. Under control conditions, 2.1–3.4% of cells initially seeded migrated through the Matrigel to the underside of the insert in 10–11 h.
In some experiments, in which SS was not applied, either an exogenous NO donor [S-nitroso-N-acetyl-penicillamine (SNAP); Sigma] or an exogenous MMP-2 inhibitor (MMP-2 inhibitor I; Calbiochem) was added. Cells were seeded onto inserts housed in companion plates as described above and incubated for 6–7 h at 37°C, 5% CO2-95% air followed by the addition of 500 μM SNAP or 10 μM MMP-2 inhibitor I for 4 h. Experimental inserts remained in the incubator throughout the duration of the experiment along with their companion controls. SNAP was diluted in cell culture medium (DMEM-F-12 + 10% FBS and 1% P/S), and MMP-2 inhibitor I was diluted in DMSO [0.5% (vol/vol) of the final culture medium; Sigma].
After SS or chemical exposure, inserts were first examined under ×100 light microscopy to ensure that the SMC monolayer was still intact and had not been compromised by SS or chemical application. The conditioned medium from the top of the insert was then removed and stored at −20°C for future assays. In some cases, samples of this medium were used to determine the number of cells that became dislodged from the GFR Matrigel as a result of SS or chemical exposure. This number was found to be minimal (<4% of total cells seeded).
Cells that had migrated through the Matrigel to the underside of the insert were fixed and stained with DiffQuik staining solution (Dade Behring). Cells on the top that had not migrated and the GFR Matrigel coating were mechanically removed with a cotton swab. The fixed cells on the underside of the insert were then counted in five ×100 fields, one in the center of the insert and four around the edges. The average number of cells migrated in these fields was used to quantify SMC migration. In some experiments, after application of SS, conditioned medium was removed and cells remaining on the GFR Matrigel layer were lysed. This lysate was then stored at −80°C and used in subsequent Western blot analyses to determine the influence of SS on PDGF-AA production (described below).
Determination of both total and active MMP-2 levels in SMC conditioned medium.
Total (latent and active) and active MMP-2 levels in SMC conditioned medium obtained from inserts containing cells exposed to either SS or SNAP were quantified with the use of a commercially available ELISA (Amersham Pharmacia Biotech) according to the manufacturer's specifications.
Determination of NO levels in SMC conditioned medium.
Transwell filter supports were attached to glass slides via the use of a silicone elastomer kit (Sylgard), and the slides were sterilized by exposure to ultraviolet light overnight. SMCs were then seeded onto the slides and grown in DMEM-F-12 + 10% FBS and 1% P/S for 4–5 days to reach confluence. Two hours before an experiment, the medium in three slides was changed to 2 ml of DMEM-F-12 without phenol red + 1% BSA (Sigma) and 1% P/S. In one slide, 100 μM l-NAME was added to determine its effect on shear-induced SMC NO production. Control, shear, and shear + l-NAME slides were moved to the hood containing the rotating disk apparatus described above, and 10 dyn/cm2 SS was applied to SMC monolayers for 4 h. For controls, the rotating disk was in place but remained stationary. After SS application, conditioned medium samples (500 μl) were collected from all three slides and stored at −20°C. Nitrite levels in samples were later determined through the use of a fluorometric assay described elsewhere (28).
Western blot analysis of PDGF-AA.
After exposure to SS as described above, SMCs remaining on top of the Matrigel layer were washed twice in PBS with Ca2+ and then lysed with the SDS extraction buffer as outlined by DeMaio et al. (8). Lysate was purified from insoluble material by centrifugation at 14,000 g for 10 min, and protein concentrations were determined through the use of a commercially available protein assay (Bio-Rad). Equal protein levels were then loaded onto 10% SDS-polyacrylamide gels under nonreducing conditions (10). After electrophoresis, proteins were transferred to Immun-Blot polyvinylidene difluoride membranes (Bio-Rad) and were blocked with 5% milk in Tris-buffered saline containing 0.1% Tween 20 for 1 h at room temperature. Membranes were then incubated with a polyclonal human anti-rabbit PDGF-A antibody (Ref. 10; Santa Cruz Biotechnology) for 2 h at room temperature at a dilution of 1:200. This was followed by a 1-h room temperature incubation in a horseradish peroxidase-labeled anti-rabbit secondary antibody (Santa Cruz Biotechnology) at a dilution of 1:2,000. PDGF-AA content on the membrane was then detected through the use of a chemiluminescence ECL Western blotting analysis system (Amersham Pharmacia Biotech), a 2-min exposure to Kodak Biomax Light film, and Scion Image analysis software (Scion).
Statistical analysis.
PDGF-AA content quantifications are normalized with respect to control. All data are presented as means ± SE. Data sets were analyzed for statistical significance by a Student's t-test with a two-tailed distribution. P < 0.05 was considered statistically significant. The Bonferroni correction, which gives a conservative significance level of P/m, where m is the number of comparisons to be performed, was used where appropriate. For example, when three groups were compared, 0.05 was replaced by 0.05/3 = 0.017 to determine significance.
Effect of SS on SMC migration.
The SMC migratory responses to various SS magnitudes and durations are shown in Fig. 1. In Fig. 1A, SMCs that had been seeded onto Matrigel-coated cell culture inserts were incubated for 6–7 h and then exposed to 1, 10, or 20 dyn/cm2 SS for 4 h. On completion of SS exposure, inserts were examined microscopically to ensure that a significant number of cells did not detach in response to SS, and then migration levels were quantified as described above. Because control conditions were the same in each experimental case (6- to 7-h incubation followed by 4-h exposure to stationary shear rod), all control values are presented together. In Fig. 1A, 83.0 ± 13.01 cells migrated per field examined under control conditions (n = 19). SMC exposure to both 10 dyn/cm2 (30.07 ± 11.99 cells migrated per field; n = 7; P < 0.05) and 20 dyn/cm2 (9.69 ± 6.22 cells migrated per field; n = 6; P < 0.001) SS for 4 h resulted in a significant inhibition of SMC migration. There was also significantly less migration in inserts exposed to 4 h of 20 dyn/cm2 SS compared with those subjected to 1 dyn/cm2 SS (56.5 ± 10.71 cells migrated per field; n = 6; P < 0.05) for 4 h.
In Fig. 1B, SMCs grown on Matrigel-coated inserts were subjected to 20 dyn/cm2 SS for 1, 2, 3, or 4 h to determine the duration of SS exposure required for significant inhibition of migration. Total running time for each group of experiments was 10–11 h. For example, if SS was to be applied for 4 h, the cells were first incubated for 6–7 h, a 7- to 8-h incubation preceded a 3-h shear exposure, etc. All control cases are again included in one data set because the same overall experimental conditions applied for each group (10- to 11-h total experiment time at 37°C, 5% CO2-95% air with no SS applied). Here, 52.68 ± 8.99 cells/field migrated under control conditions (n = 23), 41.91 ± 14.19 cells/field in response to 1 h of 20 dyn/cm2 SS (n = 7), 27.53 ± 8.06 cells/field in response to 2 h of 20 dyn/cm2 SS (n = 5), 24.33 ± 13.83 cells/field in response to 3 h of 20 dyn/cm2 SS (n = 5), and 9.69 ± 6.22 cells/field in response to 4 h of 20 dyn/cm2 SS (n = 6). Although there appears to be a consistent decreasing trend in cells migrated per field as SS duration increases, only the 4-h data set is significantly different from control (P < 0.001).
SS inhibited SMC migration both in the center of the insert, where there is relatively little shear, and the perimeter of the insert, where shear is maximum. Examining the central area of inserts, 88.32 ± 14.29 cells/central field migrated in control inserts (n = 19) and 17.5 ± 11.95 cells/central field migrated in sheared (4 h of 20 dyn/cm2 SS) inserts (n = 6). The difference between the two groups is statistically significant (P < 0.01). Also, examining the perimeter fields in the same inserts, 12.36 ± 3.75 cells/perimeter field migrated under control conditions and 1.88 ± 0.61 cells/perimeter field in response to SS. The difference is again statistically significant (P < 0.05).
Effect of SS and l-NAME on SMC NO production.
SMCs were grown to confluence on glass slides before being exposed to 4 h of 10 dyn/cm2 SS, both in the presence and absence of a NOS inhibitor (100 μM l-NAME). SMC production of NO under these conditions is displayed in Fig. 2. NO levels measured immediately before shear exposure were subtracted from values measured after shear exposure, so that only NO produced during experimentation is reported here. SMCs produced 0.517 ± 0.115 nmol NO/106 cells (n = 5) in 4 h under control conditions (no l-NAME or SS exposure), 1.853 ± 0.371 nmol NO/106 cells (n = 5) when exposed to SS, and 0.673 ± 0.215 nmol NO/106 cells (n = 5) when subjected to SS after exposure to l-NAME. SMCs produced significantly more NO when subjected to SS compared with control conditions (P < 0.05), but SMCs incubated with l-NAME before shear exposure did not produce significantly greater levels of NO compared with control, indicating that l-NAME significantly reduced the shear-mediated SMC production of NO.
Effect of NO levels on SMC migration.
To determine whether increases in NO had an effect on SMC migration, 500 μM SNAP, an NO donor, was added to inserts in the absence of SS. To replicate the conditions used in SS experiments, inserts were incubated for 6–7 h after cell seeding and SNAP was then added for 4 h. In control experiments, the same volume of medium was removed as in SNAP experiments, but it was replaced with an equal volume of fresh medium not containing SNAP. SMC migration levels were quantified after experimental completion, and the results are shown in Fig. 3 : 41.11 ± 5.43 cells per ×100 field migrated under control conditions (n = 9), and 23.64 ± 5.08 cells/field migrated in the presence of SNAP (n = 10). SNAP significantly inhibited SMC migration in these experiments (P < 0.05).
Conversely, experiments were conducted in which 100 μM l-NAME was added to inserts before exposure to 4 h of 20 dyn/cm2 SS. In control inserts, the same volume of medium was removed as in l-NAME inserts, but it was replaced with an equal volume of fresh medium with no l-NAME. During shear exposure, control inserts were treated as described in Effect of SS on SMC migration. Addition of l-NAME abolished the shear-mediated inhibition of SMC migration seen earlier, as 135.06 ± 16.61 cells/field migrated under control conditions (n = 4) and 135.97 ± 23.49 cells/field migrated when exposed to 20 dyn/cm2 SS and 100 μM l-NAME (n = 4). The difference between the two groups was not statistically significant (Fig. 4).
Experiments were also conducted to determine whether l-NAME had an effect on SMC migration in the absence of SS. l-NAME (100 μM) was added to selected inserts as described above, but in place of SS, both control and l-NAME inserts were then placed in a 37°C, 5% CO2-95% air incubator for the remainder of experimentation (9–10 h). SMC migratory activity was then quantified as described above. l-NAME in the absence of SS had no significant effect on SMC migration, as 83.5 ± 4.58 cells/field migrated under control conditions (no l-NAME; n = 5) and 85.73 ± 3.16 cells/field migrated in the presence of l-NAME (n = 5).
Effect of SS on MMP-2 production and activity.
The cumulative results of ELISA conducted on conditioned medium to determine the effects of 4 h of 10 or 20 dyn/cm2 SS on MMP-2 production and activation are shown in Fig. 5. Medium from cells exposed to 4 h of 10 dyn/cm2 SS had a total MMP-2 concentration of 13.55 ± 2.93 ng/ml (n = 4; Fig. 5A), whereas cells from companion control (no shear) experiments had a total MMP-2 concentration of 14.28 ± 2.84 ng/ml (n = 4). Medium collected from cells subjected to 4 h of 20 dyn/cm2 SS contained 11.50 ± 1.95 ng/ml total MMP-2 (n = 6), and medium from companion control experiments contained 13.76 ± 0.97 ng/ml total MMP-2 (n = 6). These data sets showed no statistically significant effects of SS on total MMP-2 production.
The effect of 4 h of 10 or 20 dyn/cm2 SS on active MMP-2 levels is shown in Fig. 5B. Four hours of 10 dyn/cm2 SS significantly (P < 0.05) reduced the active MMP-2 concentration (4.04 ± 0.19 ng/ml active MMP-2 in conditioned medium; n = 4) compared with control (4.88 ± 0.26 ng/ml active MMP-2; n = 4). Four hours of 20 dyn/cm2 SS also significantly (P < 0.01) inhibited active MMP-2 concentration (2.56 ± 0.34 ng/ml active MMP-2 in conditioned medium; n = 9) compared with control (3.74 ± 0.26 ng/ml; n = 9).
Effect of NO levels on MMP-2 production and activity.
After determining the role of NO levels on SMC migration (Figs. 3 and 4), we next quantified the effect of changes in NO concentration on MMP-2 production and activity levels through the use of ELISA. In conditioned medium collected from cells exposed to SNAP, the total MMP-2 concentration was 6.32 ± 0.31 ng/ml (n = 5), whereas the total MMP-2 concentration in control conditioned medium was 11.51 ± 0.77 ng/ml (n = 5). The difference was statistically significant (P < 0.001; Fig. 6). Conditioned medium from control cells had an active MMP-2 concentration of 2.07 ± 0.09 ng/ml (n = 5), and medium collected from SNAP-exposed cells had an active MMP-2 concentration of 1.44 ± 0.16 ng/ml (n = 5). The difference between the two data sets was also statistically significant (P < 0.01; Fig. 6).
Conditioned medium collected from inserts subjected to 100 μM l-NAME before exposure to 20 dyn/cm2 SS had a total (proenzyme + active) MMP-2 concentration of 28.82 ± 1.72 ng/ml (n = 4), and conditioned medium from companion control inserts had a total MMP-2 concentration of 27.77 ± 1.50 ng/ml (n = 4). The difference between the two groups was not statistically significant (Fig. 7). Conditioned medium from inserts subjected to l-NAME before 20 dyn/cm2 shear had an active MMP-2 concentration of 7.40 ± 0.71 ng/ml (n = 4), whereas control conditioned medium contained 7.43 ± 0.34 ng/ml active MMP-2 (n = 4). The difference was again not statistically significant (Fig. 7).
Effect of MMP-2 inhibition on SMC migration.
To further ensure that SMCs were responding to changes in MMP-2 activity, experiments were conducted in which 10 μM MMP-2 inhibitor (soluble in DMSO) was added to inserts in the same manner as described in the SNAP experiments above. The addition of MMP-2 inhibitor significantly decreased SMC migration levels as shown in Fig. 8 : 76.95 ± 7.49 (n = 7) cells/field migrated in control inserts, and 31.38 ± 11.86 (n = 6) cells/field migrated in experiments in which MMP-2 inhibitor was added (P < 0.05). We also conducted experiments to determine whether DMSO alone had a significant effect on SMC migration: 59.85 ± 19.32 (n = 6) cells/field migrated when DMSO alone was added to inserts, which was not significantly different from control (no DMSO or MMP-2 inhibitor).
Effect of SS on SMC PDGF-AA production.
Finally, we determined whether SMCs increased their production of PDGF-AA after exposure to SS. Cells were exposed to 4 h of 20 dyn/cm2 SS or to control conditions as described above. However, after completion of SS exposure, SMCs were lysed and samples were used in Western blotting procedures. A representative blot is shown in Fig. 9A. There was no significant effect of 4 h of 20 dyn/cm2 SS on the SMC production of PDGF-AA, as the PDGF-AA content in sheared cells was 95% of that in control cells (n = 3; Fig. 9B). To ensure that the presence of Matrigel had no effect on Western blots, SMCs were also grown on glass slides without Matrigel and exposed to 4 h of 20 dyn/cm2 SS, and lysates from both control and experimental conditions were run in Western blots. Again, there was no effect of SS on SMC PDGF-AA production, as sheared cells contained 97% of the PDGF-AA content of control cells (n = 3; Fig. 9B).
The primary findings of this study were that 1) fluid SS applied directly to SMCs grown on Matrigel-coated cell culture inserts inhibits their migration toward a known chemoattractant (PDGF-BB) in a dose-dependent manner (Fig. 1); 2) this inhibition is due to the NO-mediated reduction in MMP-2 levels (Figs. 2–8); and 3) 4 h of 20 dyn/cm2 SS does not affect SMC PDGF-AA production (Fig. 9). One other study examined the effect of SS on SMC migration indirectly by exposing cells to shear in culture and then transferring them to a migration assay (29), and another study applied pulsatile shear to SMCs and examined their spreading activity, which takes into account both migration and proliferation (35). The present study is the first to apply SS directly to SMCs while they were migrating in a quantifiable migration assay.
Abnormally high SMC proliferation in the media and subsequent migration to the intima are recognized as hallmark processes in IH. It has been well documented that IH is attenuated in areas of increased blood flow (SS; Refs. 5, 19, 20, 24), and the data presented here are consistent with those observations (Fig. 1). Increased flow has been shown to inhibit IH both in common carotid arteries (CCAs) denuded of their endothelium by balloon catheter injury (SMCs exposed to direct blood flow; Ref. 19) and endothelialized vascular grafts (5, 20, 24). A role for interstitial flow SS in the control of IH was suggested by experiments in which a silicone collar was placed around rabbit CCAs (9). Placement of the collar acted to heighten IH, and it was hypothesized that this was caused by the inhibition of metabolite transport due to the reduction in interstitial flow caused by the collar. Another interpretation of these data is that the reduction in interstitial flow decreased interstitial SS levels experienced by underlying SMCs, leading to increased levels of both their migration and proliferation.
The observation that SS inhibits SMC migration both in the central field of inserts where there is relatively little shear and in the perimeter of inserts where shear is maximum indicates that SS induces release of a soluble factor that is communicated between cells. Our working hypothesis was that SS elevates SMC production of NO, which in turn inhibits migration by suppressing MMP-2 levels. Support for this hypothesis was provided through the use of a NOS inhibitor, 100 μM l-NAME, which reversed the shear-mediated inhibition of SMC migration (Fig. 4), suppressed the production of NO (Fig. 2), and restored MMP-2 activity (Fig. 7). Administration of a chemical NO donor, SNAP, suppressed SMC migration significantly (Fig. 3). It is of note that SNAP has been used by others in SMC migration experiments and a drastic decrease in SMC migration levels was also observed (13).
Other work supports a role for NO in the suppression of SMC migration. Sarkar et al. (34) wounded a confluent culture of SMCs with a razor blade and observed their subsequent migration in the presence of three different NO donors. Each of the NO donors inhibited the number of cells migrating and their maximal distance from the wound edge. Marano et al. (27) used a silicone collar to induce intimal thickening in rabbit CCAs. Increased flow conditions diminished the IH caused by the collar, and this reduction was attributed to increased SMC NO production in response to elevated flow (and subsequent interstitial SS). It is also plausible that, in studies in which IH was suppressed under elevated flow conditions in endothelialized vascular grafts (5, 20, 24), the increased flow elevated EC production of NO, which in turn diffused to the underlying SMCs and suppressed their migratory activity. Although it is possible that these elevated NO levels acted to suppress SMC proliferation, it is also plausible that NO inhibited SMC migration, leading to the reduction in IH.
The effect of increased NO on MMP activity is controversial. In one study (40), MMP-2 levels were examined 3 days after placing elevated flow through left CCAs of rabbits. Active MMP-2 levels were significantly increased on exposure to elevated flow (and elevated NO produced by EC), and when l-NAME was used to inhibit NO synthesis, MMP levels fell. In another study, the endothelial NOS (eNOS) gene was transferred to SMCs and its effects on SMC migration and MMP-2 levels were examined (15). eNOS gene transfer inhibited SMC migration but, in contrast to the work discussed above, it also inhibited MMP-2 activity in conditioned medium. Addition of a NO donor (DETA NONOate) to medium also reduced MMP-2 activity. Thus it seems that NO has multiple effects on SMC MMP-2 production and/or activation, the balance of which is likely determined by the physiological and/or pathological state of the vessel.
Because MMP-2 is secreted in a proenzyme form and activated on the cell surface, we examined the effect of both SS and NO levels on the SMC production (secretion) and activation of MMP-2. It should be noted that we decided to look specifically at MMP-2 because it has been shown in other work to be necessary for rat aortic SMC migration (33) and its expression is elevated under altered flow conditions (4, 17). Palumbo et al. (29) also reported that 15 h of 12 dyn/cm2 SS downregulated MMP-2 but did not affect MMP-9 in bovine aortic SMCs. Addition of SNAP to the upper well of inserts for 4 h served to inhibit significantly both total (proenzyme + active) and active MMP-2 levels in conditioned medium (Fig. 6). Four hours of 10 or 20 dyn/cm2 shear did not affect SMC production of MMP-2 but did, however, reduce its activation (Fig. 5). It is of note that significant inhibition of both pro- and active MMP-2 levels in SMC conditioned medium was reported in response to 15 h of 12 dyn/cm2 SS (29), and therefore it seems that inhibition of pro-MMP-2 by SS (>4 h) occurs on a longer timescale than inhibition of active-MMP-2 (≤4 h). Reductions in mRNA and protein for MT-MMP, which activates pro-MMP-2 on the cell surface, were also reported after 15 h of SS but not after 3 h (29). It seems plausible that this inhibition of MT-MMP happens on a timescale of roughly 4 h (the time point measured in our work) and that it is responsible for the reduction in active MMP-2 reported here. Considering these data together, it appears that decreases in active MMP-2 levels are involved in the SS/NO-mediated inhibition of SMC migration. The reduction in total MMP-2 concentration after SNAP administration appears to have no effect on SMC migration by itself and is likely due to the larger dose used in the present work (500 μM) compared with other studies of SMC migration (10 μM; Ref. 13). We also report here decreased SMC migration levels in the presence of 10 μM MMP-2 inhibitor I (Fig. 8), providing further evidence for a role of MMP-2 in SMC migration.
Finally, we conducted experiments to determine the effect of SS on the production of PDGF-AA by SMCs. PDGF-AA has been shown to have virtually no chemotactic activity on its own (7) and to actually inhibit SMC migration induced by PDGF-BB when used in a modification of the Boyden chamber assay (21). SMC PDGF-AA mRNA expression has also been shown to be increased under both high (41)- and low (23)-flow conditions. We thus hypothesized that SMCs might produce more PDGF-AA when exposed to elevated SS levels, and this would suppress their migration in an autocrine fashion. Our results, however, showed no influence of 4 h of 20 dyn/cm2 SS on SMC PDGF-AA protein expression (Fig. 9). This is likely due to the fact that SS was applied within a shorter time frame in our experiments (4 h) than in other studies (41) that reported SMC PDGF-AA expression in response to increased flow (a minimum of 24-h exposure to modified flow conditions). Thus it is possible that SMCs do produce PDGF-AA when exposed to elevated SS, which effectively inhibits their migration, but this mechanism may operate within a longer time frame than was tested in our experiments.
In conclusion, this study provides the first direct evidence that fluid SS acts to inhibit SMC migration. The mechanism of SS action involves the NO-mediated downregulation of MMP-2 levels. Although this study broadened our fundamental understanding of the role of SS in SMC migration, the use of a more physiological three-dimensional interstitial flow model is required for improved understanding.
This work was supported by National Heart, Lung, and Blood Institute Grant HL-35549.
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
- Copyright © 2005 by the American Physiological Society


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Buffy the Vampire Slayer: Wrath of the Darkhul King
Invincibility, Infinate lifes and 9 of everything
At the main Press Start screen press
Code Effect Up, Right, Up, Left, Down, Right, Down, Left Enemies have health meters UP, DOWN, UP, DOWN, B, A You should hear a noice. Buffy starts with nine objects. You can refill them by dying and coming back. L, L , L, R, R, R, RIGHT, RIGHT You should hear a noice. Buffy will now have infinate lives. B, B, A, A, L, R, DOWN, UP You should hear a noice. Buffy will nowbe invincible
Contributed By: buffyonPS2 and Arguro.
Know Something We Don't?
You can submit new cheats for this game and help our users gain an edge.
Have We Been Fooled?
You can submit a problem report for any non-working or fake code in the lists above.


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You can now book and pay on-line securely with Sage Pay, Europe's leading payment processor.
For details of the payment security systems in use, please go to the: sage Pay Security Policy page.
For short breaks and last-minute bookings, please contact Louise directly.
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e-mail email@example.com
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Chicken, Vegetable & Pasta Soup
Be the first to review
READY IN: 1hr
Recipe by ozzygirl
Came across this from AWW website and had to try it. I am pleased to say its a keeper! The only addition I made was to add a small onion and some herbs.
See Discussion:
Ingredients Nutrition
- 40 g butter
- 1 large carrot, halved, sliced thinly
- 1 large leek, halved, sliced thinly
- 1 small onion, finely chopped
- 2 garlic cloves, crushed
- 2 slices bacon, chopped coarsely
- 500 g chicken thigh fillets, sliced thinly
- 4 cups chicken stock
- 2 cups water
- 1 cup vermicelli or 1 cup risoni pasta
- 1 teaspoon oregano
- 1 teaspoon basil
- 1 pinch chili flakes
- 1 teaspoon mixed Italian herbs
- 50 g baby spinach leaves
- salt & pepper
- parmesan cheese (optional)
Directions See How It's Made
- Heat the butter in a large saucepan and cook the onion, leek, garlic & carrot until softened (about 10 minutes).
- Add the bacon and cook for further 2 minutes.
- Add chicken, cook stirring, until the chicken is browned lightly.
- Add stock, water and herbs and simmer gently for up to 20 minutes.
- Add pasta and simmer gently for about 5-10 minutes or until pasta is tender.
- Remove from heat, stir in the spinach. Season to taste with salt & pepper.
- Serve topped with parmesan cheese.


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If you own or operate a restaurant, customer comment cards can help you discover how your patrons feel about the experience you offer. Distributing and reviewing the cards can be cost-effective; the key is knowing what to ask. If you want to improve operations based on customer feedback, be prepared for an added expense.
Things You'll Need
- Computer
- Microsoft Word
- Professional printing services
- Pens
Decide what you hope to achieve by distributing comment cards. Do you want to find out why there has been a consistent drop in revenue? Do you want to change the menu but need help deciding what items to keep? Have customers been complaining about the service and are you looking for specifics as to why? Knowing answers to these questions in advance can help you ask pointed questions and save time.
Use Microsoft Word or another word processor to type out the questions you'll ask. The title of your card can be "We welcome your feedback." Ask general questions about your customers to start such as their names, unless they want to be anonymous, and their age group. As for the survey, ask questions like: What is your favorite menu item? How often do dine at our restaurant—or other restaurants—per month? How can we improve our service?
Visit Kinko's or your trusty printer and ask them to design and print your cards. Three inches by five inches is a convenient size. If you want to spend the extra money, you can ask your graphic designer to incorporate your logo and other color images, but this is not a necessity. The more cards you print, the less you will pay per card. You can also order online at VistaPrint.com or OvernightPrints.com.
Distribute your cards. Ask your servers to deliver one card per table along with the check. Keep a stack with the maitre d or hostess, so your customers can fill them out if there is a wait.
When reviewing the comments, remember to have thick skin if you're asking questions that might invite criticism, especially if you're asking for feedback to improve your service.
Now that you have an idea of what your customers like and dislike, create a plan to implement change. This can mean offering better customer service training to your staff, or maybe you need to add a gluten-free section to your menu. Whatever the change might be, develop an organized plan to so the change process happens as smoothly as possible.


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Download Files directly to your web host server faster and easier
December 11, 2010 No comments Article
Needless to say that it's a hectic job to download and upload a file from some website to your hard drive and again upload and extract it to your server. Imagine how much time are you loosing in this process. But imagine how you can save your time by directly downloading it to your own web server.


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Has Iran stopped funding Hezbollah?
Hezbollah is allegedly facing an economic crisis over its continued role in the Syrian civil war. (AFP/File)
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Hezbollah is facing an economic crisis stemming from its involvement in the Syrian civil war, and budget cuts in Iran as a result of new president Hassan Rouhani's austerity policies, Lebanese sources say.
According to a report from the London paper Asharq Al-Awsat, Iran's Supreme Leader Ayatollah Ali Khamenei continues to fund Hezbollah through his separate budget, though the Iranian president stopped the government's flow of money to Hezbollah five months ago during the reorganization of Tehran's Foreign Ministry.
Adding to Hezbollah's problems is close monitoring of funding sources by American and European countries, the report stated. For example, on Tuesday, Germany outlawed the Berlin-based fundraising group Orphaned Children Project-Lebanon because it was found to be transfering money to Hezbollah. The sources in Lebanon said that "there are continuing American-European efforts to cut off the organization's funding."
Lebanese media also reported that European intelligence agencies have recently been cooperating in efforts to prevent the transfer of funds to Hezbollah, especially from South American and African countries. This European effort comes after several charity organizations in Europe were found to be attempting to transfer funds to the Shi'ite organization. Money transfer freezes have been placed on individual Lebanese citizens who live in Europe as well, since they have been marked as "potential routes" for transferring money to Hezbollah.
Top Headlines
- Has Lebanon lost its final chance for peace? Political implications behind Chatah's Iran missive and Saudi's military funding
- Friends of Syria call on Hezbollah to 'stop meddling' in Syria
- Iran says crisis is result of Israel's "wrong policies"
- US urges Iran to stop Hezbollah funding
- Parents scold Hezbollah, pleading to stop sending sons to Syria


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Laptop Questions? Ask a Laptop Technician for Answers ASAP!
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PC, PS Vita, PS4, Reviews, Xbox One
May 21st, 2015
by Matthew Pollesel
Get ready to die a lot — and not in a good or fun way.
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Liz's Easy Garlic Herb Butter
Recipe by Elizabeth
I enjoy going to the grocery store, buying a product that I enjoy and finding a way to make it homemade and less expensive. Sometimes being on a tight budget does not mean you have to sacrifice the quality of the food you cook. I don't cook gourmet or better all the time but sometimes simple things can really turn a simple meal into something better. You can use fresh herbs if you prefer but I just use the Herbs I have handy in my herb rack. This garlic herb recipe is something I just tried one day for kicks and now I love it. I'm never buying it in a store again and hopefully you will love it too. This garlic herb butter goes wonderfully with ANYTHING. Use it with rice, meats, potatoes, and vegetables. Enjoy!
Top Review by Karen Elizabeth
Very easily prepared and full of flavour, this was a super recipe to make up and store in the frdge to be used in all manner of dishes! My daughter actually made this, and was very excited to discover how easy it is to make a great-tasting butter! Thank you Elizabeth, this great recipe was made for PAC Fall 2011
See Discussion:2 Reviews
Ingredients Nutrition
- 2 pinches dried marjoram
- 2 pinches dried thyme
- 2 pinches dried rosemary
- 2 pinches dried basil
- 1 pinch dried bay leaf (chopped not whole leaves)
- 1 cup unsalted butter
- 2 garlic cloves, grated
- lemon juice (very very little amount)
Directions See How It's Made
- Make sure that your butter is softened. NOT MELTED.
- Put all of your ingredients in a mixing bowl and beat it until everything is mixed well and incorporated. Your butter will probably be a little creamy.
- Scrape the bowl and put your garlic butter in a container that has a lid and close the lid completely to make sure that no outside air will come inches
- Stick your dish in either the fridge or your freezer.
- I like to make this butter a day in advance so all of the ingredients can hang out. You don't have to let it sit if you need to use it immediately.
- NOTE : Keep in mind that a little will go a long way.


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Marcus Johansson is unlikely to play in the team's back-to-back games this weekend, but Capitals Coach Barry Trotz said he's "hopeful" the third-line center will travel with the team on its three-game road trip to Nashville, Minnesota and Dallas next week.
Trotz said he'd be surprised if Johansson practiced on Friday, and with the team playing matinee games on Saturday and Sunday, there won't be any opportunities for Johansson to skate with them then. Trotz said Johansson could practice with the team on Tuesday or Wednesday of next week, when Washington will be on the road.
Johansson suffered an upper-body injury about 12 minutes into the Capitals' last game before the all-star break, a 4-3 overtime loss to the Philadelphia Flyers. He didn't return to the game, and Trotz has said Johansson is "day-to-day." He hasn't been practicing with the team, but Johansson has been skating before practices, seen doing some light shooting and passing.
Trotz moved Johansson from second-line left wing to third-line center on Jan. 2, and in the eight games he played at center, he scored eight points. Through 44 games, he had 13 goals and 17 assists. With Johansson out, Mike Richards has been centering the third line of Jason Chimera and Tom Wilson.
Trotz said his expectations for the re-jiggered third line with Richards have been tempered by that trio only being together for two games.
"I would like it to have the standard or get back to the standard of when JoJo was there," Trotz said. "But he's not. … I talked to Chimmer today about the line, and there's an adjustment period. They're going to have to adjust to playing with Richie versus JoJo. JoJo will probably make a few more plays through the middle of the ice using his speed, and I think Richie is more of a grinder.
"They've just got to adjust their game, but Richie's has pretty good skills, and he's got really good hand skills. He can make plays, and you're going to start with the puck a lot with him on the ice. You've got to think about different things that you can do that make that line successful. I think they'll get that going."


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Dematic, MTEC SmartZone tenant, grows services and jobs in the U.S. with software product
HOUGHTON, Michigan (March 26, 2014) – Dematic, a global supplier of material-handling systems for factory and distribution centers, has maintained a satellite office at MTEC SmartZone's Lakeshore Center since May 2012. The Houghton location employs five Michigan Technological University student interns and 10 full-time staff who support Dematic's research and development functions at its North America headquarters in Grand Rapids, Michigan. Among major corporations that utilize Dematic's distribution systems are Amazon, Walmart, Walgreens, Abercrombie & Fitch and Toys "R" Us.
MTEC SmartZone's SmartStart program assists 35 start-ups, creates 20 new local jobs, earning Keweenaw Spark Plug award for 'Growth'
HOUGHTON, Michigan (March 14, 2014) – SmartStart, MTEC SmartZone's entrepreneurial coaching program, received the Keweenaw Spark Plug award for "Growth," during the second annual local business awards ceremony. The Keweenaw Spark Plug Awards are presented by the Keweenaw Chamber of Commerce and Keweenaw Economic Development Alliance (KEDA), along with co-sponsors MTEC SmartZone and Keweenaw Young Professionals. The event is designed to recognize local business and individuals for their efforts in 2013.
Lasalletech co-founder, Jim Northey, among leading financial trading industry experts to co-author book
HANCOCK, Michigan (February 19, 2014) – "Handbook of Financial Data and Risk Information" is a comprehensive reference for the technical, operational, regulatory and political issues in collecting, measuring, and managing financial data. Its audience ranges from financial industry practitioners and regulators responsible for implementing risk management systems, to system integrators and software firms.
Leinonen recently earned the Certified Global Business Professional credential
Jonathan Leinonen recently earned the Certified Global Business Professional credential from NASBITE International. With this achievement, NASBITE certifies that a CGBP is competent in global business management, global marketing, supply chain management and trade finance. In order to receive the designation, candidates must pass a comprehensive CGBP exam and have worked in global commerce for at least two years.
KEDA, GS Manufacturing garner awards for growing local business opportunities in the Upper Peninsula
MTEC SmartZone recognizes founding board chairman Phil Musser for 14 years of steady economic development and leadership
L'Anse Manufacturing President Mark Massicotte receives Operation Action U.P. Award of Excellence for 2013
L'ANSE, Michigan (January 22, 2014) – L'Anse Manufacturing, Inc. is one of four Upper Peninsula businesses to receive an Award of Excellence for 2013 during the annual meeting of Operation Action U.P. (OAUP) in Marquette on Tuesday, January 21.
The OAUP Award of Excellence recognizes outstanding businesses or major institutional employers who have contributed to the economic well-being of the Upper Peninsula. The awards were presented during the group's 50th anniversary meeting at Northern Michigan University.
Gov. Rick Snyder applauds MTEC SmartZone for creating 'synergy and power' across the U.P. in State of the State address
HOUGHTON, Michigan (January 17, 2014) – Exciting new developments at MTEC SmartZone were among key issues Michigan Gov. Rick Snyder discussed in his fourth State of the State address in Lansing Thursday evening.
Snyder talked about MTEC SmartZone's collaboration with Michigan Technological University, entrepreneurs and local governments to foster economic success in the Upper Peninsula. That collaboration has continued to thrive for more than a decade, leading to new developments and opportunities for job creation across the Upper Peninsula.


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At Upchurch Cooking Firewood you can see our home page http://CookingFirewood.com to see our most popular smoking wood for sale and they are often referred to as cooking wood for your grill, BBQ smoker and wood for pizza ovens. The Upchurch Cooking Firewood site gives you the options to have the wood shipped straight to your doorstep.
In this smoking wood video we give a overall glimpse into a few smoking wood for sale options. Some of our favorites are: Pecan smoking wood for sale. Pecan is a lighter wood but provides wonderful flavor. If you are new to a BBQ smoker and by the time you invest money in meat, you just want to have your meats turn out great so that is why I highly recommend the pecan wood for sale that we have. On our online firewood store, we have 1 cubic foot and 1.5 cubic foot options. These will arrive on your doorstep shortly by UPS worldwide shipping services.
Another smoking wood for sale we have is Persimmon wood for sale. It is a very underrated smoking wood because it provides a really sweet smoke, much better than apple wood. If you read on the internet it says that apple wood for sale is one of the best smoking woods but this is not the case. You can't believe everything you read on the internet. Mixing pecan wood and Walnut wood can give you a much better outcome on smoking your meats than using apple smoking wood.
Peach wood is another smoking wood for sale that is often overpriced for regular length split firewood logs. That is why we offer it in smoking wood pellets because not only is it more affordable for everyone, but peach smoking wood pellets will burn longer than regular length peach wood. The peach tree is a very slow grower in the orchards and this is why it can drive the price up higher. We have better smoking wood for sale options than peach wood. Try our peach wood smoking pellets and you will find a better outcome.
We smoke here at the Upchurch Cooking Firewood shop every Saturday and we have been getting excellent results with a mixture of Pecan wood and Walnut wood. We have used a 50% mixture of each wood and have passed this smoking wood recipe on to our customers. We have been getting excellent feedback with this smoking wood for sale recipe.
Another smoking wood for sale recipe has been to mix Persimmon wood with Walnut wood. Wow, this has been a very unique mixture that helps the meat fall off the bone and also provide some really great flavors from the walnut wood. Our persimmon wood comes from the SE Oklahoma region of the Cross Timbers. Our Walnut also comes from the Cross Timbers region where we have a plentiful array of fruit and nut trees.
GrowOKC is a Professsional Organic Horticulture Company. We have pinion wood for sale year round. #growokc
November 10, 2014
Hi Gerald in Wisconsin -
For a all around smoking wood for sale that can work in most smoking situations, I would say pecan smoking wood. I can't say it's the best because it depends on what you are cooking. For example, if you were smoking salmon the best smoking wood would be alder smoking wood. But, to fit an answer to your question, pecan smoking wood has quite a bit of versatility.
November 10, 2014
Is there a smoking wood for sale you have that you recommend the most? I'm looking for a smoking wood that can be used in smoking everything. thanks in advance
July 05, 2014
When it comes to smoking wood there are special trees that you look for. Smoking wood comes from the fruit orchards and nut groves of America. Using fruit wood is really the only option because it is sweet and has been bearing fruit for some time. If a particular tree has not grown fruit or food in general, it may not be what you are looking for. Frank, elm trees are awful for smoking meat. Actually Elm smells awful even for your fireplace. Elm is quite a shaggy and unattractive tree and although many were planted decades ago in America, they aren't the most useful trees after they die. Another type of firewood for sale we have is Pinion wood and it is not a cooking and smoking wood. Pinion is a pine gymnosperm and can really ruin a good piece of meat. So when you consider a good smoking wood, look to hickory, pecan, oak, apricot, walnut and these types of fruit and nut trees as your prime smoking wood. Thanks for your question Frank, it was a good one!
July 05, 2014
Can you use Elm or American Elm trees for smoking meat? There are a lot of trees around my house and I was curious. I wanted to ask before I ruined some meat
July 03, 2014
There are some smoking wood techniques that encourage you to soak your smoking wood chunks in water. This is really up to you. The plus side of doing this is your smoking wood will burn slower. The downside is if you are using your smoking wood chunks for your main heat source then you may not reach the temperature in your BBQ smoker that you are looking for. If you do soak your wood chunks in water, make sure you place them on a hot bed of natural charcoal. We sell 100% Mexican charcoal that will stay hot and burn long. Do not use the cheap American briquettes because they are packed full of chemicals and they will fizzle out in a few hours. So, this is a fine technique of soaking your smoking wood chunks in water first. Just make sure you have some reliable charcoal like our Mexican lump charcoal to help you keep your heat up in your BBQ smoker. Great question!
June 20, 2014
you have a great selection of smoking wood for sale. In my state we don't have a wide selection. Before you light your smoking wood do you soak it in water? There are so many smoking wood techniques, So i've heard people smoking their wood in fruit juices to make it sweeter. thanks and glad I stopped by!
© 2017 GrowOKC Pinion wood | Chiminea firewood | Cooking smoking wood chips


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We aren't particularly a musical family, yet, music plays a big part in our daily lives. My oldest daughter was a choral singer for several years and now she is in her second year of guitar lessons. My son is also one year into guitar lessons. My youngest daughter can be heard singing herself to sleep most nights and just this week she signed up to be part of the primary choir at her school.
I feel that music plays a huge part in our lives. Music can be a language all of its own and allows people to communicate even if there is a language barrier. Music can evoke emotion and memories which we may not otherwise be aware of.
When I was pregnant with all three of my children, we played classical music to my belly. I am a firm believer that those early days set in place a love and desire for music.
Coming up in February both of my guitar playing children are involved in a themed recital. Matthew is learning to play a song and his instructor is going to sing. Emily has her song ready and she is both playing and singing.
I would love to see them develop their skills and passion further, and maybe even one day attend a school of music, such as a music institute in Paris, or other destinations abroad.
I once read that music is what makes humans human, and with music we can all communicate - how very true!


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|America is Entrepreneurship
There's a couple I see every so often on my walking routes.
Together, they pick up recyclable items.
They each have tricycles and I've noticed they also have a pickup truck.
There are quite a few people I see picking up what can be recycled for cash.
This particular couple are older and they work quite hard, having to load their cycles and the materials as they move about.
When I first noticed them some time ago, they were on foot using baskets on wheels.
Not sure if they had the pickup truck then.
The tricycles are a recent addition, I think.
I guess they park the truck at a central location and make the rounds on the cycles.
It isn't a glamorous job (most jobs aren't), but I guess it makes ends meet.
It is noble and honest work, quite laborious due to the sifting, accumulating and transporting.
But I see them together, and I'm sure they also spend together what they earn together.
Many couples spend most of their waking hours apart, with one or both working somewhere away from the other.
Time apart is healthy, but a life apart when considering all things, I'm not so sure of.
Years ago when I lived with my grandmother and mother, I would gather up the recyclable items and would realize between $20 – $30 dollars for a month's worth of aluminum cans and clear plastic bottles.
I saw it as tightening a leak (not throwing money away / making the most of every opportunity).
The back seat and trunk of the car would be packed full with these items for the short drive to a recycling center.
Most cities in the Los Angeles area pick up the recyclables along with the trash (two different trucks).
When considering the small amount paid for recyclable materials, the city's effort must deal in large volume to make money and pay its employees.
There are many more individuals picking up recyclable material than city employees competing in the same effort.
On an individual level, as with this couple, the gain is very little.
The individual needs to also deal with large volumes to make it worthwhile.
I don't recycle my items anymore, but leave them the day before trash day for whoever is earning an honorable yet difficult living.
I understand the $20 a month I can get back is more appreciated and useful in the hands of such as these.
I wonder if it would be better for all parties if the city wouldn't pick up the recyclable items and instead they were available for those many who wouldn't or couldn't qualify for the few city jobs.


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Today there is a large set of coloring books. You can find coloring-cursive books, books outliners, for boys (all sorts of appliances, machines, robots, etc.), for girls (fairy-tale characters, dolls, funny funny little animals). There's even a wallpaper coloring books that opens a wide field to have enough space for young artist.
Coloring has always been children's favorite hobby. Many psychologists believe that the coloring can develop knowledge, improve motor skills and creativity at an early age.
Spider Drawing For Kids
Subject selection of images - Spider Drawing For Kids. You can use these free coloring pages for your documents, web sites, art projects or presentations. Don't forget to link to this page for attribution!
All the Images,Graphics, Arts are Copyrighted to The Respective Creators, Designers and Authors. We Don't Intend to Showcase Copyright Images, Pictures, Graphics, Arts. The Arts, Graphics available on our website are for Personal Use Only (For Desktops, Laptops, Mobiles, Tablets Screen Backgrounds). If you have any questions, please contact us.


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France has been greatly influenced due to this country's location, size, and power. France is located in Western Europe, bordering the Bay of Biscay and the English Channel. To find France on a map, the exact coordinates are 46 00 N and 2 00 E. This reasonably large country in roughly 547,030 square miles, and to better understand its size, France is slightly less then twice the size of Colorado. The terrain includes mostly flat plains, which dominates northern France, and gently rolling hills in the west. The Central Uplands lie mainly in the central and eastern area. …
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Home > Childrens Beds > Single Beds
Brand: Kidz Beds
Kidz Beds Alfemo Rosa Single Bed
Product Description:
A beautiful single bed from the gorgeous Rosa collection, the perfect bed for your little princess. Made with a high grade MDF, this bed features a French inspired design as seen by the ornamentation on the headboard and beautiful art deco legs. Complete with a pink painted finish, your little one will fall in love right away with their new bed.
Key Features:
• The materials used in the Rosa Single Bed are as follows: Melamine, Glass mirror, Matte Velvet fabric and Resin feet.
• Made from a high grade MDF and finished in a pink painted finish.
• Made to an excellent standard including beautiful floral detailing along the headboard.
• Features a French inspired design on the headboard with beautiful art deco legs.
• Matching and co-ordinating furniture available in the Rosa Collection.
About the Brand:
Kidz Beds are a European based company specializing in high quality children's furniture which will grow with your child into their teenage years.
The range features a variety of bed finishes in Natural & Whitewash, choose from Mid Sleeper Beds, High Sleeper Beds, Bunk Beds, Triple Bunks & L-Shaped Bunk Beds. This range is finished to a very high standard in terms of wood processing and manufacturing. All of the beds are made from solid wood to test the rigors of growing children. Most of the range is a modular range allowing you to convert any of the elevated beds into 2 single beds.
Be the first to Write a Review for this item!
Kidz Beds - Alfemo Rosa Bedside Locker€260.00 €209.00Buy Now
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Kidz Beds - Alfemo Rosa Mirror€250.00 €199.00Buy Now


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This Weeks Top Treats
This week at Outlawz Challenges on Twisted Thursday the twist is SCALLOPED EDGES! The challenges are always ANYTHING GOES, but with the f...
Today, I have been working on another pair of crochet Sneakers! Finished this pair in gray!
I love making these adorable diaper cakes, also called Nappy Cakes. This particular one is a custom order. My customer, who also happen...
Sunday, January 3, 2016
Embroidered Monogrammed Handkerchief
After a couple trial and errors making this design I finally had one turn out perfect! My Dad has always been the guy to have a hanky in his pocket. So I thought what would be more fitting than a custom monogrammed hanky?
First off I ran out of bobbin thread during my first attempt so after I changed the bobbin I had issues with the loopity loo's and how thick the thread was coming out onto the hanky versus the way it went so smoothly on the first try. I found out that on my SE-400 Embroidery machine I had put my bobbin in upside down! I even looked at the picture drawings and thought I was doing it right. The great thing about Facebook there are a lot of groups to ask questions so I posted this picture and asked what am I doing wrong. That is when I found out that my bobbin may be in wrong. Flipped that baby over changed out the thread just to have the same color spool as the bobbin. I was so pleased to see that it was not the machine.
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It really is simple to find people's belief regarding how to put money into the investing in gold sector as absolutely everyone includes a diverse angle on what to expect during the inventory market place at each level in time, but almost all of the time people's belief may be incredibly puzzling. Quite possibly the most typical dilemma that new buyers do have is ways to decide fantastic investments with the undesirable types, what to speculate on, what time to make investments among the other people. A lot of the thoughts you need to respond to to be able to create a fantastic determination whenever you desire to commit are highlighted beneath.
Is that this an excellent Time to Put money into Shares?
Around the off probability that you are getting a gander at dollars markets amid a lofty reduce, chances are you'll assume it is a terrible time to commence investing. Around the off likelihood that you are having a gander at it when shares are reviving, you could consider it can be a decent time.
Neither one of the situations is basically good or awful in the event that you might be investing with the long haul (ten years or more). Nobody can anticipate with any level of assurance which way the share trading method will move at any provided time; nonetheless more than the extensive haul, stock markets has continually moved better. Every bear advertises is trailed by a buyer market (when inventory charges increase). Verifiably, positively trending marketplaces have endured any longer than bear markets, plus the additions of buyer markets have more than counterbalance the misfortunes in bear marketplaces
Exactly how much Danger Really should I Choose?
A standout among quite possibly the most important fundamentals of investing will be the cozy connection among chance and returns. Devoid of hazard, there is often no income. You ought to will to simply accept a lot more risk over the off possibility that you're seeking for more noteworthy returns. In that regard, chance is often some thing to be grateful for, however just in case you take into consideration suitable the perfect time to enable the inescapable market cycles occur. By and enormous, in case you might have a far more drawn out enterprise time skyline, you ought to will to hope a far more noteworthy evaluate of danger, to the grounds that there'll be additional prospect for the market to work by the in this article and there cycles. Typically, comprehending economical professionals have been compensated with optimistic long haul returns.
New investors are often inspired to put essentially in frequent money, which could give minute enhancement, supplying one of the most excellent approach to lessen possibility. By putting resources into a couple of different shared assets chatting with various useful resource classes, (for example, expansive growth stocks, world shares or bonds), you'll be able to reduce unpredictability substantially promote without having yielding lengthy haul returns.


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Do yourself a favour don't eat at the restaurant. There was cats walking around the place, ie; jumping on chairs and trying to get on tables. We should have left then. We waited for 10 minutes for a menu after I went and asked for...More
Getting you more information on this room
Getting you more information on this room
"Room 57 with the outside bathroom, air cond and comfy bed."
"Do the upgrade and get the rooms with outdoor bathrooms they are soooo nice :)"


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Now PlayingLexus F SPORT Line of High Performance Luxury Cars
Lexus is known for its reliable luxury cars, but the brand is also capable of putting out some pretty high class performance. The Lexus F SPORT lineup is where most of the...Read More
PlayLexus LFA Supercar Overview & Ride-Along Track Test with Scott Pruett
Autobytel had the opportunity to attend the Lexus #WhatstheF media event at the Las Vegas Motor Speedway, which Lexus organized to bring attention to the...Read More
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Play2014 Lexus ES 350 Test Drive & Luxury Car Video Review
Lexus is somewhat alone in the mid-size luxury sedan marketplace in that it offers a choice of two 3.5-liter V6 sedans. For sporting oriented drivers, the marque presents...Read More
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When Autobytel gets a chance to test drive the 2013 Lexus LS 460, a new and interesting feature gets most of the attention. This Autobytel video focuses on the LS...Read More


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Events: First Annual Minds Behind the Music Summit
- At July 5, 2009
- By NESHA
- In events
Imperial Marketing Group will host the First Annual Minds Behind the Music Summit on Saturday, July 11 at PNC Studios, 10 Jay Street, Suite 206, Brooklyn, NYC at 1:00 p.m.
Minds Behind the Music will provide exclusive insider tips and advice on how to pursue a career in the music and entertainment industries within various fields such as marketing, publicity, music production, journalism and more. The program will feature a candid panel discussion, Q&A session, and conclude with a networking reception. The goal of the Summit is to reach high school and college students, and introduce them to a side of the business not often represneted by the media.
Featured panelists include:
Amy "Aimstar" Andrieux
Senior Editor/ General Manager, The Source Media Group
Amanda "Precise" McIntyre-Chavis
Executive of Mechanical Licensing/ Artist Development, EMI/ Capitol Music Group
Shukmei Wong
Co-Founder/ Publicist, WeRoqq Publicity & Promotion
Anslem Samuel
Music/ Entertainment Journalist
Popular Culture & Entertainment Journalist/ Souleo Enterprises, Inc.
Justin Henry
Studio Engineer/ Producer, Chung King Studios
A King
Program Director/ On-Air Personality, PNC Radio
Jake Paine
Editor and Chief of HipHopDX.com
Artist/ MC
One comment
Sup everyone,
Thanks for the support! You're all definitely welcome to come out and network. Hope to see you all Saturday!July 7, 2009


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BASEBALL; Fade to Black: Pinstripes in Peril on WPIX
By RICHARD SANDOMIR
Published: September 2, 1993
Free telecasts of Yankee games, a tradition for 43 years, seem headed toward extinction because WPIX and the MSG Network are at an impasse.
"This is what the consumer feared all along, the loss of all free games to cable," said Michael Eigner, the president of WPIX/Channel 11. "I've been hopeful of making a deal, but they've made an offer and said, 'Take it or leave it.' "
Control of all Yankees telecasts has resided with MSG since 1991 as part of its $486 million cable contract with the team. Over the last three years, MSG has aired 75 games, paid about $5 million a season for the time on WPIX to telecast 50 games and then sold the advertising for both outlets. MSG said it was now time for WPIX to pay for the right to show Yankee games and has proposed a figure of $15 million annually. Mets Have Even Split
When MSG and WPIX struck their deal before the 1991 season, the reason was the importance of keeping a significant portion of games on over-the-air, or free television. MSG had 4.9 million subscribers, including 3.5 million in the New York market.
The cable network is available in 49 percent of the TV homes in which WPIX is seen in in the New York market area, so appearing on WPIX expanded the geographic reach of the games. Appearing on free TV enables viewers who can't afford cable or can't get it to watch games. The Mets have an even split of telecasts, with 75 games on SportsChannel and 75 on WWOR/Channel 9.
MSG officials say that they have lost $22 million on the WPIX arrangement and that continuing it is economically unfeasible. WPIX says MSG is making a profit, but MSG refuses to include in its WPIX arithmetic the advertising revenues and subscriber fees it gets for its cable games.
"It's a tough situation," said Douglas Moss, president of the MSG Network. "All we're trying to do is make a reasonable deal. But we can't continue to suffer losses."
During the spring, MSG proposed that WPIX continue to air 50 games a season through the year 2000 for a $15 million annual fee. At $300,000 a game, said Moss, "that's exactly what we're paying now, not one cent more." He was referring to MSG's contract with the Yankees, which costs about $5 million a year.
WPIX rejected the offer, and made three counterproposals, said Eigner: continue the current arrangement; a revenue-sharing deal in which WPIX would sell the advertising time and split revenues with MSG; pay the $15 million rights fee if MSG shared one-third of its subscriber fees.
Last week, MSG rejected the counteroffers. Moss called them all "fancy ways for us to keep losing money."
"They have not offered anything else," said Eigner. "There have been no other proposals from them. They said they'd accept any reasonable offer and we made three of them." Additional Talks Planned
Moss said they would sit down for talks again later this month.
"We have months and months to work on this and come up with something new," he said.
"We don't have to offer WPIX anything, but we don't want to hoard games," said Bob Gutkowski, president of Madison Square Garden, the parent company of the MSG Network. Both are part of Paramount Communications.
Eigner said: "The Bronx and Brooklyn have very low cable penetration. Over 80 percent of the homes don't subscribe to cable or can't get it. So we want to keep the franchise on the station."