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Resistance exercise (RE) training, designed to induce hypertrophy, strives for optimal activation of anabolic and myogenic mechanisms to increase myofiber size. Clearly, activation of these mechanisms must precede skeletal muscle growth. Most mechanistic studies of RE have involved analysis of outcome variables after many training sessions. This study measured molecular level responses to RE on a scale of hours to establish a time course for the activation of myogenic mechanisms. Muscle biopsy samples were collected from nine subjects before and after acute bouts of RE. The response to a single bout was assessed at 12 and 24 h postexercise. Further samples were obtained 24 and 72 h after a second exercise bout. RE was induced by neuromuscular electrical stimulation to generate maximal isometric contractions in the muscle of interest. A single RE bout resulted in increased levels of mRNA for IGF binding protein-4 (84%), MyoD (83%), myogenin (∼3-fold), cyclin D1 (50%), and p21-Waf1 (16-fold), and a transient decrease in IGF-I mRNA (46%). A temporally conserved, significant correlation between myogenin and p21 mRNA was observed (r = 0.70, P ≤ 0.02). The mRNAs for mechano-growth factor, IGF binding protein-5, and the IGF-I receptor were unchanged by RE. Total skeletal muscle RNA was increased 72 h after the second serial bout of RE. These results indicate that molecular adaptations of skeletal muscle to loading respond in a very short time. This approach should provide insights on the mechanisms that modulate adaptation to RE and may be useful in evaluating RE training protocol variables with high temporal resolution.
- neuromuscular electrical stimulation
- myogenin
- p21
- MyoD
it is generally accepted that resistance exercise (RE) elicits adaptations that result in increased skeletal muscle size and changes in contractile characteristics. The majority of these adaptations take the form of increases in myofiber size (e.g., Ref. 20) and alterations in the mix of contractile and metabolic proteins present in these cells [e.g., isoform shifts, etc. (14)]. Alterations in the types and amounts of cellular proteins in myofibers can involve changes in gene transcription and translation. In vitro and in vivo studies have identified a number of specific mRNAs in skeletal muscles that change in response to increased loading (15, 18, 21, 24, 37, 40).
The concept underlying RE training, designed to induce skeletal muscle hypertrophy, is that the correct selection of program variables, e.g., frequency, intensity, and duration of individual sessions, results in cellular and molecular responses that promote the growth process. We have shown that individual sessions of high-force actions common during an RE session elicit cellular and molecular responses in rodents that have characteristic temporal patterns of increase and decline (24). In those studies, we also found that subsequent sessions of high-force actions result in the summation of these responses. The magnitude of summation is sensitive to the time interval allowed between RE sessions (24). In a subsequent study, we found that molecular responses to high-force RE sometimes differ between skeletal muscle of spinal cord-injured and ambulatory individuals (13) but are generally similar.
In vitro and in vivo studies indicate that two primary processes are involved with the compensatory hypertrophy response in mammalian skeletal muscles. The first are anabolic processes necessary for the accretion of protein to support myofiber enlargement. The second involves proliferation of satellite cells that appear to be necessary to provide myonuclei to the enlarging myofibers (7, 41, 42, 45–47, 49). The cellular and molecular mechanisms underlying these processes have been extensively characterized in a number of cell types, particularly in cell culture settings. In particular, the steps associated with the initiation of cellular proliferation are generally well established (e.g., can be found in cell biology textbooks). Based on these well-characterized processes, we have previously used representative mRNAs as markers of potential myogenic responses to high-force actions of skeletal muscle in rodents and humans (2–4, 13, 24). For example, increased expression of myogenin is often associated with committed stages of myoblast differentiation and fusion (22, 27, 44, 48, 53). Similarly, the cyclin-dependent kinase inhibitor p21 is associated with withdrawal from the cell cycle and, often, subsequent cell differentiation (22, 33). Together, these markers are often used as an indication of myogenic cell differentiation in settings associated with muscle formation or muscle hypertrophy (2, 4, 22, 24, 33). Potential alterations in cellular proliferation can be inferred from changes in key cell cycle regulatory agents such as cyclin D1 (38). In the present study, we used this myogenic marker approach to characterize the molecular level responses of human skeletal muscles to acute bouts of RE (maximal isometric contractions).
Following a protocol similar to that used in our previous rodent studies, we found that RE induced similar molecular level changes in human skeletal muscles in response to a single exercise bout (24).
The in vivo validation of loading-sensitive cellular and molecular responses will help to elucidate the processes that have been assumed to take place as a result of RE training but, until now, have not been systematically investigated. This understanding will, in turn, allow for the further elucidation of the regulatory mechanisms that control and coordinate the suite of adaptations that comprise the compensatory hypertrophy response and, at least in a laboratory setting, provide a mechanism by which different RE training protocols and modalities (e.g., static vs. dynamic, concentric vs. eccentric, etc.) could be objectively evaluated.
Nine (age 25 ± 4 yr, height 178 ± 8 cm, weight 70 ± 3 kg, means ± SE; 6 men, 3 women) able-bodied subjects participated in this study. The subjects had no history of lower extremity pathology and signed informed consent before testing. Subjects were recreationally active and not currently involved in lower extremity RE. The methods were approved by the Institutional Review Boards of The University of Georgia, Shepherd Center, and the University of California-Irvine.
RE protocol.
Neuromuscular electrical stimulation (NMES) was chosen to ensure maximal activation of the vastus lateralis (VL) muscle and to avoid the complication associated with voluntary muscle activation (e.g., recurrent inhibition, motor unit cycling, etc.). It has previously been demonstrated that this method of muscle activation compares favorably with voluntary muscle contraction for the development of muscle hypertrophy and increased strength (e.g., Ref. 43). The VL muscle was stimulated, as described previously (5, 16, 29, 43). Subjects were seated in a custom-built chair with the hip and knee secured at ∼70° of flexion. The leg was firmly secured to a rigid lever arm with an inelastic strap to ensure that the knee extensors could only perform isometric contractions. As with our previous animal studies, isometric mode contractions were used to minimize the potential for skeletal muscle injury (24). The moment arm was established by placing a load cell (model 2000A, Rice Lake Weighing Systems, Rice Lake, WI) parallel to the line of pull and perpendicular to the lever arm. Torque was recorded from the load cell by using a MacLab analog-to-digital converter (model ML 400, ADInstruments, Milford, MA) sampling at 100 Hz and interfaced with a portable Macintosh computer (Apple Computer, Cupertino, CA). Two 8 × 10-cm surface electrodes (Uni-Patch, Wabasha, MN) were placed on the skin over the proximal and distal portions of the VL muscle. Using magnetic resonance imaging, our laboratory has previously demonstrated that this electrode placement will maximally recruitment primarily the VL muscle (6, 29). A commercial stimulator (TheraTouch model 4.7, Rich-Mar, Inola, OK) was used for NMES. The initial torque was determined in the following manner. Subjects performed a maximum voluntary contraction (MVC) for isometric knee extension before NMES. The subjects were highly motivated, and all had prior experience with performing knee extension MVC. We have previously established that the VL muscle constitutes roughly 30% of the total quadriceps group cross-sectional area (28). Accordingly, NMES current sufficient to elicit ∼30% of the observed isometric knee extension MVC (generated by the entire quadriceps group) was determined and used for the subsequent NMES protocol. This 30% index value provides a means to avoid the unnecessary delivery of excess current that may activate synergistic muscles that were not the focus of this study. The NMES protocol consisted of 5-s contractions separated by 15 s for 30 min at these previously determined current levels. Contractions were evoked with 50-Hz trains of 450-μs biphasic pulses.
Biopsy technique.
Biopsies were taken from the VL by using the percutaneous biopsy technique, as done previously (16, 28). To reduce the number of serial biopsies, the preexercise sample was collected from the nonexercised muscle. To minimize the potential for interference, any serial biopsy samples were collected at least 2 cm from previous biopsy sites. Samples were immediately cooled with liquid nitrogen and then stored at −70°C until analyzed.
Experimental timeline.
See Fig. 1. At time 0, a biopsy sample was collected from the VL of the nontreatment leg. This approach was used to ensure that any changes seen in the 12-h biopsy samples could not be attributable to a previous biopsy procedure. The use of the contralateral limb as a control has been previously validated (e.g., Ref. 35). The subjects then completed the initial 30-min bout of NMES-induced RE. At 12 h postexercise, a biopsy sample was collected from the VL of the exercised leg. At 24 h, a second biopsy was collected from the previously exercised VL, and the NMES-induced RE exercise was then repeated. Biopsy samples were then collected from the exercised VL 48 and 96 h post-NMES-induced RE, relative to time 0 (24 and 72 h after the second bout of RE).
Total RNA isolation.
Measurements of total RNA content provide insights on the translational capacity of tissue. Total RNA was extracted from preweighed frozen muscle samples by using the TRI Reagent (Molecular Research Center, Cincinnati, OH), according to the company's protocol, which is based on the method described by Chomczynski and Sacchi (17). Extracted RNA was precipitated from the aqueous phase with isopropanol and, after washing with ethanol, dried and suspended in a known volume of nuclease-free water. The RNA concentration was determined by optical density at 260 nm (using a 260-nm optical density unit equivalent to 40 μg/ml). The muscle total RNA concentration is calculated based on total RNA yield and the weight of the analyzed sample. The RNA samples were stored frozen at −80°C to be used subsequently in determining specific mRNA expression with the use of relative RT-PCR procedures.
One microgram of total RNA was reverse transcribed for each muscle sample using the SuperScript II RT from GIBCO BRL and a mix of oligo(dT) (100 ng per reaction) and random primers (200 ng per reaction) in a 20-μl total reaction volume at 45°C for 50 min, according to the provided protocol. At the end of the RT reaction, the tubes were heated at 90°C for 5 min to stop the reaction and then were stored at −80°C until they were used in the PCR reactions for specific mRNA analyses (see below).
A relative RT-PCR method using 18S as an internal standard (Ambion, Austin, TX) was applied to study the expression of mRNAs for IGF-I, mechano-growth factor (MGF), IGF-I receptor, IGF binding proteins (BP)-4 and -5, myogenin, cyclin D1, and p21. The sequence for the various primers used for the specific target mRNAs has been published previously (13). Primers were designed using Primer Select computer program (DNA Star), purchased from Life Technology, GIBCO, and were tested for their compatibility with the alternate 18S primers.
In each PCR reaction, 18S ribosomal RNA was coamplified with the target cDNA (mRNA) to serve as an internal standard and to allow correction for differences in starting amounts of total RNA.
For the 18S amplification, we used the Alternate 18S Internal Standards (Ambion), which yields a 324-bp product. The 18S primers were mixed with competimers at an optimized ratio that could range from 1:4 to 1:10, depending on the abundance of the target mRNA. Inclusion of 18S competimers was necessary to bring down the 18S signal, which allows its linear amplification to the same range as the coamplified target mRNA (Ambion, Relative RT-PCR kit protocol).
For each specific target mRNA, the RT and PCR reactions were carried under identical conditions by using the same reagent premix for all of the samples to be compared in the study. To validate the consistency of the analysis procedures, at least one representative from each group was included in each RT-PCR run.
One microliter of each RT reaction (0- to 10-fold dilution, depending on target mRNA abundance) was used for the PCR amplification. The PCR reactions were carried out in the presence of 2 mM MgCl2 by using standard PCR buffer (GIBCO), 0.2 mM dNTP, 1 μM specific primer set, 0.5 μM 18S primer-competimer mix, and 0.75 unit of DNA Taq polymerase (GIBCO) in 25-μl total volume. Amplifications were carried out in a Stratagene Robocycler with an initial denaturing step of 3 min at 96°C, followed by 25 cycles of 1 min at 96°C, 1 min at 55°C (55–60°C depending on primers), 1 min at 72°C, and a final step of 3 min at 72°C. PCR products were separated on a 2–2.5% agarose gel by electrophoresis and stained with ethidium bromide, and signal quantification was conducted by laser scanning densitometry, as reported previously (52).
The RT and PCR methods used in this work have been extensively validated in previous publications (13, 52). Each specific mRNA signal is normalized to its corresponding 18S. For each primer set, PCR conditions (cDNA dilutions, 18S competimer-primer mix, MgCl2 concentration, and annealing temperature) were set to optimal conditions, normalized so that the target mRNA product yields were in the linear range of the semilog plot when the yield is expressed as a function of the number of PCR cycles, and, for a given condition, 18S and target mRNA PCR yields were tightly correlated to input cDNA.
Statistical analysis.
All values are reported as means ± SE. Treatment effects were determined by ANOVA with repeated measures by using the Prism software package (Graphpad). Pearson's correlation analysis was used to assess the relationship between p21 and myogenin by using the Prism software package. For all statistical tests, the 0.05 level of confidence was accepted for statistical significance.
Analysis of the torque output records indicated that, over the two exercise sessions, the subjects produced 29 ± 3% of their previously measured isometric MVC during the NMES-induced high-force actions. The NMES current used to induce these isometric actions was 151 ± 15 mA.
Components of the IGF-I axis.
Expression and/or accumulation of mRNA for IGF-I was decreased by ∼46% 12 h after a single session of NMES-induced high-force isometric actions (Fig. 2A). The level of MGF mRNA, the loading-sensitive isoform of IGF-I (25), was unchanged at all time points (not shown). The mRNA for IGFBP-4 was increased by 84% by a single session of NMES-induced high-force isometric actions, but this response decayed such that it was no longer elevated 24 h postexercise (Fig. 2B). The imposition of a second session of NMES-induced high-force isometric actions 24 h after the first resulted in an additional increase in IGFBP-4 mRNA to approximately twofold above the preexercise value that was sustained through 96-h postexercise (Fig. 2B). There were no changes in the levels of IGFBP-5 or the IGF-I receptor mRNAs at any time point (not shown).
Myogenic regulatory factors.
The expression and/or accumulation of MyoD mRNA was increased by 83% 12 h after a single session of NMES-induced high-force isometric actions (Fig. 3A). The level of mRNA for myogenin was increased 2.8- to 3.1-fold at 12, 24, and 48 h postexercise (Fig. 3B).
Markers of cellular proliferation and differentiation.
The mRNA for cyclin D1, a non-muscle-specific marker of cellular proliferation, was increased by 50% at 12 h after the first session of NMES-induced isometric actions (Fig. 4A). The mRNA for p21, a general marker of cellular differentiation, was increased 16- to 20-fold in the postexercise samples at 12, 24, and 48 h postexercise (Fig. 4B). There was a positive relation between myogenin and p21 mRNA when analyzed by individual data point (Fig. 5A) or by time point (Fig. 5B).
Total RNA concentration.
Total muscle RNA increased 96 h postexercise (Fig. 6).
On a cellular level, the implicit aim of RE training programs, designed to induce skeletal muscle hypertrophy, is the optimal activation of the anabolic and myogenic mechanisms that result in the accretion of the proteins necessary to support an increase in myofiber size. Because these cellular-level mechanisms must be activated to achieve meaningful adaptations, their activation must precede these adaptations. As a matter of course, studies that involve the development of measurable adaptations, such as increased skeletal muscle or myofiber size, will not provide information on the dynamic state that exists as the adaptation processes are initiated.
In this study, we have used previously established molecular markers, representative of several cellular-level myogenic processes, to elucidate the time course of the response of human skeletal muscle to acute sessions of high-force isometric actions with a temporal resolution of hours rather than weeks. In contrast to voluntary muscle actions, NMES results in consistent, maximal activation of all effected myofibers with each stimulation. The design of the present study was closely patterned on our previous work conducted in rodents in an effort to translate those findings from the rat model to humans (3, 24).
Potential weaknesses.
Some compromises were made during the translation of the experimental design from rodents to humans. Specifically, a second session of high-force isometric actions was performed in the same subjects after just two single-session time point samples were obtained. This was done to avoid having to use a second cohort of subjects to complete this study. However, this approach sacrificed the potential to observe the decline in responses over time. In addition, fewer time points were used to reduce the number of biopsy samples that each subject was required to provide. Due to the size of the biopsy samples, the number and type of analysis targets were also reduced. Due to this limitation, we focused on mRNA-level analyses, which provided the greatest number of data points for a given tissue sample.
The high-force muscle contractions are common in RE and, as used in this study, were very similar to that used in our laboratory's previous work with rat and humans (3, 13, 24). As with our rodent and human studies, NMES was used to avoid the complications associated with voluntary contractile activity, including the problem of motor unit cycling, which would increase variability when using biopsy sampling (3, 13, 24). These foundational studies used isometric as opposed to dynamic muscle actions to reduce the possibility of skeletal muscle injury, which might tend to complicate the interpretation of results. While isometric-mode resistance training is currently less common, it has been shown to be effective in producing increases in muscle size and strength (e.g., Refs. 30, 36, 43).
IGF-I axis.
IGF-I has been shown to stimulate anabolic and myogenic processes associated with the development of skeletal muscle hypertrophy (1). In skeletal muscle, the IGF-I system has also been shown to be sensitive to increased loading (1, 2, 4, 24). In this tissue, the IGF-I axis consists of locally expressed IGF-I and MGF, the type I IGF-I receptor, and a number of IGF-I binding proteins. Modulation of the various components of this system appears to be important for a compensatory hypertrophic response (1, 8, 9, 12, 19, 23, 25, 32, 51). In humans, RE has been variously reported to increase (Ref. 11; lengthening muscle actions), decrease (39), or not change (13, 26) IGF-I mRNA levels. We had previously reported that our acute RE protocol resulted in increased IGF-I mRNA in rodents (24). In contrast to that result, in the present study, IGF-I mRNA was found to decrease by almost 50% at the earliest time point (Fig. 2A). Using a different exercise program (e.g., 250–350 dynamic contractions), Psilander et al. (39) also found that IGF-I mRNA was modestly decreased 1–6 h postexercise. We had previously found that IGF-I mRNA was unchanged 24 h after two bouts of RE in humans, which would agree with our current findings (13). In animal studies, increased skeletal muscle loading has been shown to result in an upregulation of the expression of IGFBP-4 and downregulation of IGFBP-5 (8, 24). In the present study, we found that the mRNA for IGFBP-4 was increased 12 h after a single bout of RE and then started to return toward control by 24 h. The second bout of RE conducted after the 24-h time point resulted in a further increase in the mRNA for IGFBP-4. In a possible contrast, Bamman et al. (11) found that RE involving lengthening skeletal muscle actions resulted in a decrease in IGFBP-4 mRNA. Unlike the results from rodent studies, we did not find that the mRNA for IGFBP-5 was depressed following RE (8, 24). Also, in contrast to our previous rodent studies, we did not see an increase in MGF mRNA following RE (24). In agreement with this result, we had previously found that two bouts of RE resulted in an increase in MGF mRNA in spinal cord injury but not control subjects (13).
Myogenic regulatory factors.
The myogenic regulatory factor (MRF) family is made up of a set of transcription factors that determine the myogenic fate of cells. Although various members of this family may have overlapping functions, specific roles for the various MRFs have been proposed. MyoD is most often associated with cell determination and is often found to be highly expressed in actively proliferating myoblasts and may be critical for proliferation in response to stimuli such as skeletal muscle injury (34, 44, 48, 50). Increased expression of myogenin is often associated with committed stages of myoblast differentiation and fusion (22, 27, 44, 48, 53). Alternatively, myogenin expression may occur within myofibers and may be associated with alterations in metabolic properties (31).
In the present study, the expression and/or accumulation of MyoD mRNA was modestly increased 12 h after the initial bout of RE (Fig. 3A). However, the subsequent bout of RE did not stimulate an increase in MyoD. Psilander et al. (39) also reported a transient increase in MyoD mRNA immediately following their RE bouts.
The mRNA for myogenin experienced a fairly robust increase in response to RE that was maintained through the 24-h time point (Fig. 3B). The second bout of RE resulted in a renewed or continued elevation of myogenin mRNA that decayed by the final 96-h time point (i.e., 72 h later).
Cell proliferation and differentiation.
In contrast to the MRFs, markers such as cyclin D1 and cyclin-dependent kinase inhibitor p21 are generalized or nonskeletal muscle cell-specific markers of potential proliferation (cyclin D1) or differentiation (p21; p21 is a cell cycle inhibitor). In rats, we found that cyclin D1 mRNA was relatively insensitive to a single bout of RE but responded with an approximately twofold increase following a second RE bout (24). In the present study, we found that cyclin D1 was increased only at the earliest post-RE time point and only by ∼50% (Fig. 4A). An elevation of cyclin D1 expression would suggest an increase in the number of cells within the previously exercised muscles that are activated to enter the cell cycle in response to this stimulus. Visual comparisons between the cyclin D1 and MyoD data appear to suggest that these two message pools were responding in a similar manner (Figs. 3A and 4A). However, there was no significant correlative relationship between these two data sets (data not shown).
The mRNA for p21 demonstrated the most dramatic magnitude of change of any of the markers analyzed for this study (Fig. 4B). The cyclin-dependent kinase inhibitor p21 is associated with withdrawal from the cell cycle and, often, subsequent cell differentiation (22). Because there did not appear to be a high signal indicating proliferative activity, the increased expression of p21 mRNA suggests that a number of cells within the exercised skeletal muscles were most likely going through the process differentiation. As our laboratory has reported in a number of previous studies involving increased skeletal muscle loading (4, 13, 24), there was a strong correlation between the mRNA for myogenin and that of p21 (Fig. 5A). Further analysis indicated that this relationship retained temporal coherence (Fig. 5B). These results strongly suggest that at least some of the cells going through the process of differentiation were myogenic precursor cells.
Total RNA.
The majority of the RNA present in skeletal muscle cells consists of ribosomal RNA. Therefore, the concentration of RNA in skeletal muscle provides an indication of the synthetic potential of the cells that make up that tissue. However, it should be noted that potential differences in translational efficiency would not necessarily be reflected in this measurement. In a previous study, we found that the concentration of RNA in skeletal muscle of spinal cord injury subjects before the RE bouts was lower than that in skeletal muscle of able-bodied control subjects but that, after exercise, the RNA concentration was not different between these two groups (13). In skeletal muscle from the able-bodied subjects, two sessions of RE did not result in any changes in RNA when measured 24 h postexercise, a result that is similar to that seen in the present study (13). However, herein we report that two sessions of exercise did induce an increase in RNA at the 96-h time point (72 h after the second RE bout). This suggests that the anabolic potential of these skeletal muscles was increasing after just two sessions of RE but with a substantial delay relative to the molecular markers.
Exercise mode.
The question arises whether the exercise mode used in this study may account for differences with published results using dynamic muscle actions. For example, in one such study, Bamman et al. (11) had subjects complete eight sets of eight squat exercises on two different occasions. In one session, shortening muscle actions were performed; in the other session, lengthening actions were performed. Muscle biopsies were collected 48 h postexercise in each case. These authors reported that the mRNA for IGFBP-4 was decreased and that of IGF-I increased in response to lengthening contractions conducted at 110% of one repetition maximum (11). Compared with the present results using isometric RE (Fig. 2), this indicates that lengthening muscle contractions may lead to a difference in adaptation, either in magnitude or time course. However, it should be noted that, at the 48-h time point, following the lengthening but not shortening contractions, these subjects reported high levels of soreness, had reduced MVC, and had serum creatine kinase levels that were approximately threefold higher than baseline measurements. Therefore, it is not clear whether the changes seen were related to nascent adaptive responses or to muscle injury.
In a more recent study, Bamman et al. (10) measured the expression of MRF and cell cycle-related proteins in the muscles of young and old subjects of both genders, before and after dynamic-mode RE. In that study, the RE consisted of three sets of three different dynamic REs designed to recruit the quadriceps muscle group for shortening or shortening plus lengthening muscle contractions (10). In biopsy samples collected 24 h postexercise, no changes were detected in protein levels of the MRFs or the cyclins measured. These authors did observe a decrease in the levels of cyclin-dependent kinase inhibitor p27 in female subjects postexercise. In the context of the present study, it is not clear whether these results represent RE mode-specific differences or a latency in the translation of MRF and cyclin mRNAs.
Our laboratory recently reported that, following resistance training programs that resulted in muscle hypertrophy in rats, significantly elevated levels of MGF and IGF-I mRNA could be detected in the muscles of animals that had been trained using either isometric or shortening but not lengthening muscle contractions (3). However, due to the experimental design, it cannot be determined whether these results represent mode-specific responses to the last bout of RE or a different steady state induced by 10 serial bouts of RE training.
Perhaps the most compelling argument for a mode-specific difference in molecular level responses to RE comes from the work published by Hameed et al. (26). In that study, subjects completed 10 sets of knee extensor exercise at 80% of one repetition maximum with muscle biopsies of the VL collected at 2.5 h postexercise. These authors report that RE induced an increase (range, 2–864%) in the mRNA for MGF in the muscles of young but not old subjects, whereas no RE-induced change in IGF-I mRNA was detected. Whereas the results presented herein do not include a postexercise time point before 12 h, the findings published by Hameed et al. may suggest that exercise mode plays a role in determining the pattern of molecular level responses to RE.
In summary, the results of this study indicate that a single session of RE is sufficient to stimulate molecular-level responses, which is indicative of the initiation of myogenic processes in skeletal muscle. It is important to note that the RE session used in this study consisted of a total of 90 electrically stimulated contractions with a total duty cycle of 7.5 min of contractile activity. Just two bouts of this activity, a total of 14 min of contraction spread over 2 days, was sufficient to induce an increase in the concentration of total RNA, most likely representing an increase in ribosomal RNA, and, therefore, an increase in the anabolic potential of this tissue. These results, along with a number of other published observations, indicate that the mechanisms that regulate the adaptation of skeletal muscle to increased loading respond on a relatively short time scale and that these responses may be useful in evaluating RE training variables with high temporal resolution.
Funding was provided, in part, by the Foundation for Physical Therapy (C. S. Bickel) and National Institutes of Health Grants HD-37439-S, HD-39676, and HD-39676-S2 (to G. A. Dudley) and AR-45594 (to G. R. Adams).
The authors thank the subjects for their participation, particularly their willingness to provide multiple biopsy samples. We also acknowledge Anqi Qin for technical assistance with the molecular analysis.
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 the American Physiological Society


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Making a Quilt Sandwich
Quilters call the process of assembling the three layers--quilt top, batting, and backing--as making a quilt "sandwich."
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Reviews // 2 Articles
Summer Movie Preview: June 2011
2 May 2011 // 5:00 PM
The return of everyone's favorite mutants, the arrival of an intergalactic peacekeeper, and some guy with a bunch of birds marks June's idea of sunny Summer fun.
'The Art of Getting By' Suffers from Artists' Block
8 Jan 2012 // 4:03 PM
Little of this movie's inertia can be blamed on the actors when their characters are summarized, not dramatized – and the screenplay forces them to do most of the summarizing themselves.
'The Art of Getting By': Find Something
17 Jun 2011 // 12:39 AM
Everyone has heard this story before. And that's the trouble with The Art of Getting By.


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text: I've been observing my hair and on many strands, at different points, there's a crinkle. The crinkle is generally about a centimeter long and feels rough and looks discernibly different from the rest of the strand. These crinkles are at various points of the strands; most, however, are near the ends. They really mess with any sort of curl pattern and I'm not really sure where they came from. I'll try to post a pic because I did a search and it seemed like someone with relaxed hair had a similar problem but people said it was their hair type. This is not about my hair feeling wiry or dry or hard. It's about these magical crinkles so unless hair type changes sporadically throughout a single strand, I highly doubt that these crinkles are related to my hair type. Has anyone else dealt with these before? Does anyone know where they come from? It seems like I might have to cut to get rid of them (which I am not ready to do) but are there any temporary solutions?


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VirnetX Holding Corp. (VHC) CTO & CSO Robert D Iii Short sells 20,000 Shares
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CTO & CSO of VirnetX Holding Corp. (VHC) Robert D Iii Short sells 20,000 shares of VHC on 01/18/2012 at an average price of $25.94 a share.
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FAQ Translations
What do you mean when you say Macalester is a "liberal arts" college?
A liberal arts education provides students with a broad foundation of knowledge in the natural sciences, mathematics, the social sciences, the humanities and the fine arts. Students also focus on a field of study (a "major") and demonstrate mastery of the intellectual building blocks and methodologies of that academic discipline. In contrast to an education that emphasizes narrow technical or vocation training, a liberal arts education will develop the critical thinking and communication skills that will equip graduates to adapt to a rapidly changing world.
What degrees does Macalester offer?
Macalester College offers programs leading to the Bachelor of Arts degree with majors in 38 areas of study as well as 37 minors and 10 concentrations. Students are not required to officially declare a major until the end of their second year. For detailed information on areas of study go to Academics at Macalester.
Is it possible to receive credit from Macalester for International Baccalaureate, A Level and/or Advanced Placement exams?
For students admitted to Macalester through the first-year admissions program, Macalester will accept credit for no more than the equivalent of twenty Macalester credits from courses earned prior to matriculation at Macalester. (The only exception to this limitation applies to students who have achieved the equivalent of twenty-four Macalester credits solely through College Board Advanced Placement Examinations.) Credits earned through Advanced Placement Exams, International Baccalaureate or GCE A-Level examinations cannot be used to meet the college's general distribution requirement.
Students whose scores on Higher Level Examinations of the International Baccalaureate Program are 5, 6, or 7 may receive the equivalent of four or eight semester hours of credit for each such examination. Credit is determined by the appropriate academic department on an individual basis. No credit is awarded for the Standard Level Examinations.
Students who have completed A-Level Examinations in order to earn a General Certificate of Education (GCE) receive the equivalent of eight semester credits for each A-Level Examination with a score of A, B, C, or D. No more than the equivalent of twenty semester credits can be earned in this manner.
Students who have taken the College Board Advanced Placement Examinations may be eligible for advanced placement and appropriate credit. Scores of 3, 4 or 5 may result in credit. Academic departments have determined which scores result in credit. More information on Transfer of Credit. Some departments require the student to discuss their scores with the department chair before a credit determination is made. First year students who wish to have their AP scores considered should have them sent directly to Macalester College by the College Board. You may visit the AP Exams website at http://www.collegeboard.org/?excmpid=VT-00026/ or call 888-225-5427 to request your scores be sent directly to Macalester College – use 6390 for the college code.
Students who have studied at international secondary schools that use similar exams (CAPE, French Baccalaureate etc.) may also receive credit based on exam results. The final determination on eligibility for advanced standing based on such exams is made by the Registrar.
How does Macalester provide academic advising to its students?
Each student has one faculty member designated as their primary advisor. The advisor is responsible for monitoring the progress of the student through their career at Macalester. In a student's first year, they will register for a First-Year Course as one of four first-semester classes. First-year courses are for first-year students only, are limited to sixteen students, and the instructor of each course is the academic advisor for the students in that course. Students are encouraged to change advisors to someone in their primary academic area of interest at the end of the second year.
Application and Selection
What does Macalester look for in deciding which international students will be offered admission?
The competition for admission to Macalester is very keen, as we receive over 1,200 international applicants each year for the approximately 60 spaces reserved for international students in each class. We are seeking students who have the potential to thrive in our rigorous academic program and will also be engaged and contributing members of the Macalester community.
Our review of your application will begin with a thorough analysis of your academic preparation and secondary school performance. School systems vary a great deal and we always look at an applicant's high school record in the context of the system in which they were educated. Whatever the system, we look for evidence that the applicant has performed at a very high level academically and has taken a rigorous curriculum. The standard requirement for transcripts submitted to Macalester is that they include grades (marks) for years 9 through 12. Because there are multiple educational systems around the world with unique grading structures, however, we waive the transcript requirement on a case-by-case basis. Many other things are considered as we evaluate an applicant's potential to be a successful and engaged member of the Macalester community, including personal qualities, experiences and background, talents, etc. We will look for that information in your application, essays and recommendations, among other things.
Does Macalester require that applicants submit SAT exam scores?
Applicants to Macalester must submit results from either the SAT or ACT exams. These standardized exams are often very different from the exams that international students have taken previously, so Macalester does not have a minimum score nor do we rank our applicants on the basis of their SAT or ACT scores. Register for the SAT on the College Board website. Macalester's code for the SAT is 6390.
Are SAT Subject Exam (SAT2's) required?
Macalester does not require that students submit results from SAT Subject exams (sometimes referred to as SAT2 exams). However, many international students choose to submit the results of SAT Subject exams because these results may demonstrate their knowledge and ability in those academic areas.
Are all international applicants required to submit TOEFL exam results?
International applicants are required to submit either TOEFL or IELTS results in order to demonstrate proficiency in English. International applicants whose first language is English and/or who attend secondary schools where the primary language of instruction is English may request that the TOEFL requirement be waived. To register for the TOEFL, visit http://www.ets.org/toefl. Macalester's code for the TOEFL is 6390.
What is the minimum TOEFL score required for admission?
Macalester does not have a minimum TOEFL or IELTS score requirement for admission. However, because Macalester's classes are taught in English and we do not offer any English-as-a-second-language courses, only students who have a strong command of English language (reading, writing and speaking) will be offered admission. Most international students who are offered admission to Macalester have scored 100 or higher on the TOEFL iBT (600 or higher on the paper-based TOEFL) or 7.0 or higher on the IELTS.
What is Macalester's minimum SAT score?
Macalester does not have a minimum score requirement for the SAT, nor does it rank applicants on the basis of SAT scores. The value of the SAT results varies significantly from case to case and it is always only one piece of information among many that we use in evaluating an applicant's potential to thrive academically at Macalester.
If I take the SAT and/or TOEFL in December, will those results be considered as part of my Regular Decision application?
Any SAT and/or TOEFL exams results received by the third week in January will automatically be considered as a part of your Regular Decision application. We recommend that you request a "rush report" of the results if you take the exams in December.
Must I submit official SAT and TOEFL reports directly from the College Board or ETS?
We understand that it can be very expensive to have official score reports sent to all of the colleges to which you are applying. We will accept a photocopy of your exam results as long as it comes from an appropriate school official who verifies that the results are correct. If we have any questions about the legitimacy of the scores, we will contact the school official.
If I have been taking O Level / A Level type curriculum, are A Level results required?
Students who have only completed the O Levels will not be considered for admission to Macalester. A Level students must submit final O, AS and A Level results for evaluation. If final A Level results are not available by the application deadline, predicted scores must be submitted instead. This requirement also applies to students who have been educated in countries with similar educational systems.
Should my high school transcripts and letters of recommendation be submitted in the original language or translated into English?
Every applicant must submit official transcripts with the school seal stamp and official signature sent directly by a school official. If the transcript is not in English, the applicant must also submit English translation of the transcript with the name and position of the person who translated the document. If we have any concerns about any school documents, we will contact the school in order to verify the authenticity of the credentials.
My school does not have a "guidance counselor" so who should complete the Secondary School Report?
Your high school principal, head of school or other designated school official may complete this recommendation form. This individual should identify his or her official role at the school. The form should not be completed by an agent or anyone other than a school employee.
If I apply online, can I submit my academic credentials by mail? Must all application materials be submitted at the same time?
You have the option of applying online or downloading the application forms and mailing them in. If you apply online you may still submit other materials through the mail. You may submit all of your documents at once or as they become available. Make sure that each document includes your name as it appears on your application.
My mid-year grades will not be available by the Regular Decision application deadline – will that be an obstacle to my admission?
Secondary schools around the world run on a wide range of academic calendars and in many cases the mid-year grades may not be available by the application deadline. Students should submit the mid-year grades as soon as they become available, preferably by the end of February. If that is not possible, email us at firstname.lastname@example.com and explain when they will be available.
May I submit supplementary materials with my Macalester application?
In most case the required application materials are sufficient for an applicant to convey the information we need in order to evaluate their candidacy. However, we welcome additional writing samples, recommendations and other items if you feel they will supplement the information in your application in any important way.
Is it possible to pay the Macalester application fee by credit card?
Yes. When you submit your application to Macalester online, The Common Application will prompt you to pay the $40 fee, which you can do with a credit card. If you are unable to pay the fee, click "Request a Fee Waiver." You will need to ask your school counselor to submit a waiver request. For more information, see next question.
Will Macalester waive the application fee?
Macalester automatically waives the $40 application fee for students who submit Part 1 of the application online by November 15. In all other cases a student who wishes to have the application fee waived should write a letter or send an email to firstname.lastname@example.com explaining why the application fee would present a financial hardship for you and your family. Waiver requests that are endorsed by an appropriate school official are most likely to be approved.
Are admissions interviews required? What is the purpose of an interview?
Admissions interviews are not required. Many of our international applicants do not have an opportunity to have an interview, so we do not penalize applicants who have not interviewed. At Macalester the interview is a brief and relatively casual conversation which is intended to help us begin to get to know you as a person. It is not an exam or audition, and you will not be given a grade or score at the end of the interview.
How do I decide whether to apply Early Decision or Regular Decision?
Applying Early Decision (E.D.) to Macalester is appropriate if, after extensive research about your college options, you decide that Mac is the college that will best meet your academic and personal goals. As an E.D. applicant you are committing to withdraw applications to other colleges and enroll at Macalester if offered admission and (if applying for financial aid) your family finds it affordable. In order to be considered for admission under an E.D. process you must submit the Early Decision Agreement, signed by you, a parent and a high school official by the application deadline.
There are three possible outcomes to an E.D. application: you may be offered admission, you may be denied admission or a decision on your application might be "postponed" to the Regular Decision round. We sometimes postpone E.D. applicants to the Regular Decision round because we want to have the opportunity to compare the applicant to the students who apply in the Regular Decision pool, or because the student requires a substantial financial aid award and we do not yet have a clear picture of how many large scholarships we will be able to offer. We often end up offering admission in the Regular Decision round to students who were postponed from E.D., so a student who is postponed should not give up hope. However, the student should keep applications to other colleges active in case they are not offered admission to Macalester. We use the same methodology to determine eligibility for financial aid in both decision periods, so you would receive the same award no matter which process you used.
Financial Aid
I will need financial assistance in order to be able to attend Macalester – does that hurt my chances of admission?
Macalester's broad international representation is made possible in great measure by a significant institutional commitment to providing financial assistance to international students. Approximately 80% of the international students who enroll at Macalester received financial aid awards. We are committed to meeting the full demonstrated financial need of every student we admit. Unfortunately, our financial aid budget is not unlimited, so the amount of financial aid an applicant requires is a factor as we decide which students will be offered admission, and students requiring large scholarships will face greater competition in the selection process.
How much financial aid does Macalester offer?
Macalester is committed to meeting the full demonstrated need of every enrolling student. In 2012-2013 Macalester provided $9.1 million in financial aid for international students.
Does Macalester offer "merit scholarships" to international students?
The Charles J. Turck Presidential Honor Scholarship is awarded to a limited number of international students who demonstrate special potential to excel academically and contribute significantly to the Macalester community. The scholarships range from $10,000 to $15,000 per year. Every international applicant is automatically considered for the Turck Scholarship – no special application is required.
When will I hear if I will receive financial aid from Macalester?
Students who are offered admission to Macalester will receive information about any financial aid they have been awarded at the same time they are notified of the admissions decision.
What are the components of a financial aid package?
It is difficult for us to give you a specific number for the amount of aid you would receive. We review the financial aid paperwork that you submit to us and if you are admitted you receive an award along with your admittance letter. Macalester is committed to meeting the full demonstrated need of every enrolling student. In 2012-2013 Macalester provided $9.1 million in financial aid for international students. Below is further information about the financial aid package.
A financial aid package from Macalester College will usually include:
Grants (gift aid): Grants made up more than 80% of the financial aid distributed by Macalester in 2011-2012. The vast majority of these grants and scholarships come from Macalester, primarily funded by gifts from alumni and friends of the college. Grants do not need to be repaid.
Student Employment: Students typically have the option to work approximately 9 hours per week on campus and earn about $2000 over the course of the year.
Student Loans: Small loans are usually a component of an international student's financial aid package. Repayment of the loan does not begin until six months after you cease being a Macalester student, and students may request that the payments be deferred if they go on to graduate school. In some cases, students who return to their home country after graduation may request that the loan debt be forgiven. Our experience is that most of our international students are able to get jobs upon graduation and pay off the loan without much difficulty since they have 10 years in which to complete payment.
If I have questions that are not answered by the Macalester website, who should I contact?
We strongly encourage prospective international students to complete our online Information Form. This will make it easier for us to share information with you quickly and efficiently. If, however, you have a question that has not been answered on our website or by the communications we send you once you have filled out the online inquiry form, please email us at firstname.lastname@example.com. Include your full name, country of citizenship and your high school in your email.
If I don't have any additional questions, why should I get on the Macalester mailing list?
Getting on our mailing list not only makes it easier for us to provide you with important information about Macalester, it will make it possible for us to notify you if an admissions officer or other member of the Macalester community will be in your area.
If I do get on the Macalester mailing list, how will you communicate with me?
Our experience is that the most efficient and reliable means of communicating with international students is through email. If you prefer to receive hard copies of some of materials, you may request them by emailing firstname.lastname@example.com.
Is it possible for me to contact a current Macalester student?
On the Admissions & Financial Aid page of the Macalester website there is a section called "ASK A STUDENT" which you may use to communicate directly with a current Macalester student. If you would like to communicate with a current student from your country, please email firstname.lastname@example.com.
How "international" is Macalester?
Macalester is one of the most culturally diverse communities you will find at any undergraduate college in the United States. In each entering class approximately 12% of the students are international and the student body includes citizens of more than 90 countries. Macalester's American students are often bi-cultural or have spent a significant amount of time living in a country other than the U.S. The academic program also is infused with many opportunities to study other languages, literatures, histories, religions and political systems.
How do international students adapt to being part of the Macalester community?
Macalester's international students come from a wide variety of backgrounds and each student's transition is unique. The College has a strong support system in place to help students as they adjust to life in the U.S. and at college. Key among these support systems is the department of International Student Programs. It is worth noting that 95% of the international students who enroll at Macalester complete their degree.
Do all Macalester students live on campus?
All first- and second-year students are required to live on campus. Students have the option of moving off-campus for junior and/or senior year.


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CREATIVE SCRAPPERS #81This dear little baby bird sat on my lawn and didnt move! I ran inside and grabbed my camera and some baubles, placed them carefully behind it and it STILL didnt move!!! How lucky lucky lucky!!! I got some beautiful photos! Anyway it was perfect for the sketch over at Creative Scrappers, you can find it here
STUCK? SKETCHESYou'll probably get sick of 'Alice' photos...still.... they need scrapping!! Over at Stuck? Sketches, there was a triple photo sketch for this week and good for what was needed here!!! I had fun with this! The title reminds me of the 'lessons' and era from where Alice was trying to be taught to help her 'grow up'.


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Pontiac Problems? Ask a Mechanic for Answers ASAP
Hello and thank you for using Just AnswerI'm Paul and I will help you with your Question
when the security light blinks this is because the computer is have a problem reading the ignition key through the passlock sensor.
you can attempt a relearn of the passlock by turning the key to the crank position for one second. then release to the run (on) position
leave the key in the on position for 12 minutes
the theft light should go out.
once the theft light goes out try and start the vehicle. if it does not start on first try turn key to off and then retry.
you may need to repeat the 12 minute procedure up to 3 times to get it to relearn
if it will not relearn you will need to replace the passlock sensor
let me know how you make out and if you need more help


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Speak softly, carry a big book: County Recorder Darlene Maneval to retire at end of year
She may come off as mild-mannered and soft-spoken, but Becker County Recorder Darlene Maneval is passionate about keeping good local service at the courthouse.
And Maneval, who is retiring at the end of this year, also feels strongly about keeping elective county positions like the recorder, and the auditor-treasurer, elective, and not appointed by the county board.
Good local service is important to Maneval. She has been known to drive into Detroit Lakes in a blizzard, on a Sunday, to help a stranded travelor in Florida who needed a birth certificate.
But that good service provided by the Recorder's Office to farmers and smaller banks (in the form of lien searches on personal property via the Uniform Commercial Code) is threatened by Minnesota Secretary of State Mark Ritchie, who Maneval says has arbitrarily decided to cut county recorders out of the system and require small banks and farmers to go online for state services.
That will cost Becker County about $9,000 a year in lost fees, but more importantly, will make life difficult for people who should be able to go to the courthouse for help as they have always done, she says.
"The ones that will be really hurt by this are small local banks and farmers," who often need the lien checks on equipment that serves as collateral prior to planting and harvesting.
"They can get it here and take it to the bank the same day," she said.
The state plans to cut counties out of the system as of Dec. 1.
Maneval and other county recorders plan to fight it.
When it comes to elected versus appointed county department heads, it's an easy decision to Maneval.
It's a matter of checks and balances, she believes, and there are times elected officials like the recorder and auditor-treasurer have to make difficult stands.
Roots run deep
Maneval has more than 30 years under her belt at the Recorder's Office, where she was first hired in 1981.
Back then, Eleanor Miller ran the Recorder's Office, and hired Maneval as a fill-in for her deputy, who went on maternity leave.
"I must have impressed her, because I'm still here," Maneval joked.
Miller, who went by E.T. Miller, perhaps to downplay her gender, started in the Recorder's Office in the 1930s and became county recorder in 1955, when her predecessor, Oscar P. Morken, died in office. Miller also died while in office.
Thankfully, Maneval's predecessor, Marlene Martinson, broke that unhappy trend when she retired 12 years ago.
And Maneval, who replaced her, is now 62 and looking forward to retiring young enough to enjoy herself.
She'll join her husband, Steven, who retired 18 months ago from the U.S. Fish & Wildlife Service in Detroit Lakes, where he was head of maintenance and handled prescribed burns and wetland restoration.
One hobby she plans to pursue in her new-found spare time is genealogical research.
"I love it. It's my favorite part of the job – genealogy research," she said. To that end, she plans to volunteer at the County Historical Society.
From big books to digital
The Recorder's Office has always been known for its big books of documents, but like everywhere else, it has gone digital over the last 10 years or so.
The basic elements of the job haven't changed, though: "Foremost, we have all land records, abstracts and torrens (registered land thorough the court system)," dating back to 1870 and 1871 when the county first started with handwritten deeds.
The recorder's office also handles vital statistics – birth, death and marriage certificates. But not divorces. The court administration office handles those.
The five-person office also handles passports and Uniform Commercial Code work, which brought in $165,000 over the past 10 years.
Playing catch-up
And the office has been busy digitalizing past records. "We now have records complete to 1978," she said. Deeds and mortgages are digitalized back to 1970, but the associated documents are not yet finished.
In fact, all documents back to 1952 have been digitalized. "The image is there with a document number, and we're now seeking bids to get the index and associated information completed," she said.
The money for digitalization comes from a $10-per-document fee, and several companies specialize in the work, contracted via the bidding process.
But Maneval has been saving money by having her and her staff do the work in their spare time.
"In the frugal line, I thought that would be good," she said with a laugh. "But now we're short-staffed and we don't have time … I've got 46,607 documents to be indexed – I don't think I can do 46,606 documents in six months."
She'll bid the work out, and make a decision based on "who has a good reputation and a good price."
In all, there are 617,399 documents in the Recorder's Office, and roughly half have been digitalized so far.
Full primary slate
When it comes to her replacement, there will be a crowded primary election ballot, with names that include Maneval's chief deputy (Karen Wenner), her vital statistics administrator (Sandee Listul), the head of the county planning and zoning department (Patricia Swenson), and the chief deputy in the Hubbard County Recorder's Office (Darryl Hensel), who lives in Becker County.
"There are four people running for my office and I fully endorse all four of them," Maneval said. "Each and every one could do a good job." She has full confidence in the voters of Becker County to make the right choice.


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Twelve Places to Celebrate Día de Muertos in New York
Gran Electrica, Fonda, and Salvation Taco are throwing parties.By Cristina Alonso
Gran Electrica, Fonda, and Salvation Taco are throwing parties.By Cristina Alonso
Day of the Dead foods can be found all over Chicago.By Michael Gebert
Here's more reasons to dine in the burbs, eat game, and get in touch with autumn harvest.By Collin Keefe
Restaurants get in on the spirit of Halloween.By Collin Keefe
Four activities to keep you from robbing fruit from off our relatives' graves.By Hadley Tomicki
The L.A. Times' food blog jumps the gun a little on L.A. food writers' favorite holiday.By Hadley Tomicki
With Halloween over already, what's a ghoul to do?By Collin Keefe
Seven celebrations and one drink recipe from The Varnish help the concept of death go down a little smoother.By Hadley Tomicki
Where to eat during Day of the Dead/Dia de los Muertos.By Hadley Tomicki
Boerum Hill: Now that Trout has taken over Gravy, the new menu is not surprisingly chock-full of fish dishes. [Bergen Carroll] Midtown East: Richard Sandoval's restaurants including Pampano are celebrating Day of the Dead through November 2 with specialty foods like pan de muertos, which is a sweet bread that's baked to look like a skull! [Grub Street] Midtown West: At convivial spot La Bonne Soup, which has been a French staple on West 55th Street since 1974, you can have a soup, salad, bread, dessert, and a glass of wine for $16.75, only slightly more than half the price of lunch at Jean Georges. [Lost City] Nolita: D.J. Mark Ronson has designed a playlist specifically for Noble Food & Wine. Dinner gets it own soundtrack. [Down by the Hipster] Tribeca: A Murray's Bagel partner is opening Original Zucker's Bagels & Smoked Fish on Chambers Street at the end of November. [NYS] Upper West Side: The Triad Theater is now promoting a new musical called Wine Lover. The audience will sample six wines throughout the performance. [Snack]
New Yorkers Are Getting a 5-Story Chick-fil-A That 'Pays Homage' to 9/11
BYGGYZ Sandwich Shop Opens Today at Noon
The Absolute Best Bar on the Upper West Side
Nom Wah Tu Wants to Serve Late-night Dim Sum
USDA Warns That Millions of Pounds of Fake 'Organic' Imports Are Pouring Into U.S.


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In September we started the upgrade of our vehicle fleet, buying 7 new Ford Focus ECOnetic cars. With a fuel economy of 74.2 mpg and CO2 emissions of 99 g/km, the Focus ECOnetic was chosen to support our aim of reducing the environmental impact of our operations.
DRB runs a fleet of 58 vehicles including 41 vans, 14 cars and 3 wagons. Whilst recognising the need for these vehicles in delivering our services to our customers, we also recognise our responsibilities in terms of reducing the impact of our operations on the environment; and where we see opportunities we are starting to look for ways to operate in a more sustainable fashion. The choice of these ECOnetic cars was one such opportunity.


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Sept. 14 for Balfour meeting with Junior class during 3rd
period – Class Rings
Sept. 28th & 29th for class ring
orders during lunch
September 9, 2017
October 28, 2017
December 9, 2017
February 10, 2018
April 14. 2018
June 9, 2018
86days until End of 1st Semester
Principal: Travis Mayfield; Assistant Principals: Chip Bevis - Athletic Director, Kevin Dawson, Jason Franklin, Missi Oakley, Ranesa Shipman
The Wilson County School System does not discriminate
due to age, race, color, gender, national origin, disability, religion, creed,
genetics or veteran status in the provision of services, in programs or
activities, or in employment opportunities or benefits. The lack of English
language skills will not be a barrier to admission and participation in
educational programs. Translation services are available at each school and at
the Central Office. Inquiries concerning Title VI or Title IX of the Civil Rights
Acts should be directed to Chuck Whitlock at (615) 444-3282. Inquiries
concerning Section 504 and IDEA should be directed to Dawn Bradley at (615)
453-3282. Inquiries concerning the American with Disabilities Act should
be directed to Mickey Hall at (615) 444-3282. Complaint Managers are available
at each school/location to address employee concerns.
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text: Show Hide image Politics 28 September 2011 Austrian hackers publish police data A group of Austrian hackers release police data in protest against privacy bill Sign up for our weekly email * A group of hackers in Austria have published personal data of nearly 25,000 police officials. The group known as AnonAustria has claimed that it was protesting against a bill that would oblige telephone and internet companies to store six months worth of information in order to present it to the authorities if need be. According to the BBC, the published information, which contained dates of birth and addresses, is said to have been leaked to the group rather than hacked. In a statement, the Austrian police said they were concerned, that information could endanger the named officers. Subscribe from just £1 per issue More


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Where Are We Drinking?
The canned pineapple juice-based Blue Hawaiian is the Friday happy hour special at this diviest of dive bars. But the bar tries to keep things classy by serving it in a wine glass and using real blue Curaçao -- and even including a cherry if you ask for one.
Think you know where we're drinking this week? Leave your best guess in the comments section below.
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There are many things in life Al Roker enjoys: weed, giant snakes, videobombing Savannah Guthrie, pooping his pants in the White House, y'know, normal people stuff. But there's one thing he doesn't like: Drake, or at least going to Drake concerts, where he's surrounded by loud YOUTHS and flashing lights, because #TooOld.
I'd feel the same way if a white dude wearing plaid was shouting Drake lyrics behind me.


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Skerne is a village in the East Riding of Yorkshire, England. It now forms part of the civil parish of Skerne and Wansford. The village is situated 1 mile (1.6 km) to the south of the River Hull and the Driffield Canal. It is approximately 2 miles (3 km) south-east from Driffield and 2 miles north-east from Hutton Cranswick.
Historically, Skerne was an ecclesiastical parish in the wapentake of Harthill. From 1894 until 1974, Skerne was located in Driffield Rural District. The merger of Skerne with the neighbouring civil parish of Wansford occurred in 1935.
In 1974 most of what had been the East Riding of Yorkshire was joined with the northern part of Lincolnshire to became a new English county named Humberside. The urban and rural districts of the former counties were abolished and Humberside was divided into non-metropolitan districts. The new organization did not meet with the pleasure of the local citizenry and Humberside was wound up in 1996. The area north of the River Humber was separated into two "unitary authorities"—Kingston-upon-Hull covering the former City of Hull and its closest environs, and the less urban section which, once again, named itself the East Riding of Yorkshire.
Research Tips


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Don't miss out on a lower price on this hotel
Get an overview of this hotel4-star hotel near the beach
Popular property highlights
Situated in La Linea de la Concepcion, this beach hotel is within 3 mi (5 km) of Victoria Stadium, Casemates Square, and Rock of Gibraltar. Cathedral of the Holy Trinity and The Convent are also within 3 mi (5 km).
Hotel Features
This smoke-free hotel features a restaurant, an outdoor pool, and a health club. WiFi in public areas is free. Other amenities include a coffee shop/café, a seasonal outdoor pool, and a business center.
Room Amenities
All 82 rooms offer free WiFi, minibars, and plasma TVs with satellite channels. Guests will also find room service, premium bedding, and separate bathtubs and showers.Information missing or incorrect? Tell us!Opens in a new window
AC Hotel La Linea by Marriott
Hotel Amenities
Hotel Amenities
AC Hotel La Linea by Marriott features an outdoor pool and a health club. The hotel offers a restaurant and a coffee shop/café. Business-related amenities at this 4-star property consist of a business center and a meeting room.
Public areas are equipped with complimentary wireless Internet access. Event facilities measure 646 square feet (60 square meters). This beach hotel also offers a library, a terrace, and tour/ticket assistance. Parking is available onsite for a surcharge.
AC Hotel La Linea by Marriott is a smoke-free property.
- Conference space size (feet) - 646
- Conference space size (meters) - 60
- Breakfast available (surcharge)
- Free newspapers in lobby
- Airport transportation - drop-off (surcharge)
- Business center
- 24-hour front desk
- Safe-deposit box at front desk
- Laundry facilities
- Television in common areas
- Elevator/lift
- Coffee shop or café
- Tours/ticket assistance
- Dry cleaning/laundry service
- Luggage storage
- Garden
- Outdoor pool
- Total number of rooms - 82
- Number of floors - 4
- One meeting room
- Terrace
- Restaurant
- Self parking (surcharge)
- Smoke-free property
- Concierge services
- Health club
- Free WiFi
- Library
- Multilingual staff
- Year Built 1992
- Number of buildings/towers - 1
- Outdoor seasonal pool
Family Friendly Amenities
- Outdoor pool
- Free Wi-Fi
- Laundry facilities
Available in all rooms: Free WiFi
Available in some public areas: Free WiFi
Self parking (surcharge)
Room Amenities
- Premium bedding
- Air conditioning
- Minibar
- Daily housekeeping
- Hair dryer
- Iron/ironing board (on request)
- Free WiFi
- In-room climate control (air conditioning)
- Turndown service
- Designer toiletries
- Private bathroom
- Free toiletries
- Separate bathtub and shower
- Room service (limited hours)
- Desk
- Blackout drapes/curtains
- Satellite TV service
- Plasma TV
Where to Eat
Buffet breakfasts are available for a surcharge and are served each morning between 7 AM and 10:30 AM.
AC Lounge - This gastropub specializes in Spanish cuisine and serves lunch, dinner, and light fare.
Room service (during limited hours) is available.
Nearby Things to Do
Recreational amenities at the hotel include an outdoor pool and a health club.
Hotel Policies
Check-in time ends at 3 PM
Check-in time starts at 3 PM
Check-out time is noon
Payment types
Children and extra beds
- Children are welcome.
- Rollaway/extra beds are not available.
- Cribs (infant beds) are not available.
- Pets not allowed (service animals welcome)
You need to know
Extra-person charges may apply and vary depending on property policy.
Optional extras
- Fee for buffet breakfast: EUR 11 per person (approximately)
- Airport shuttle fee: EUR 5.00 per vehicle
- Self parking fee: EUR 9 per day
Hotel Name
- AC Hotel Linea Marriott
- AC Linea
- AC Linea Marriott
- Hotel Linea
- Linea AC
- Linea Hotel
- AC Hotel Linea Marriott La Linea de la Concepcion
- AC Linea Marriott La Linea de la Concepcion
- Ac Hotel La Linea
- Ac Hotels La Linea
We should mention
Cash transactions at this property cannot exceed EUR 2500, due to national regulations. For further details, please contact the property using information in the booking confirmation.


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Buick Repair Questions? Ask a Mechanic for Answers ASAP
Hello and welcome to JustAnswer
Unfortunately you are correct Regals are notorious for a few different electrical issues, luckily we've had plenty of time to narrow them down to a couple common causes.
And what might those be?
the first step for a complaint like this is to remove the turn signal bulbs and check the contacts for corrosion, loose connections, water, other contaminants.After you have verified that is not the issue, you are forced to choose between one of three options:1) because the fault is intermittent and will not blow the bulb immediately, performing a proper and thorough electrical test on the system and guaranteeing a correct diagnosis is either extremely time consuming or not possible. option one would be to spend possibly countless hours moving from circuit to circuit in the vehicle testing wiring for excessive amperage draws and poor contact, not the best option IMO
2) replace a module called a daytime running light module, this would be my go to option, because generally when these module fail they leave behind a "burnt" smell and an visible "char" inside of them so you can take apart your old one to "confirm" your repair.
3) replace the turn signal multi-function switch ( the stalk on the steering wheel that turns on and off your turn signals ( less common than the daytime running light module, but these are known to burnt up on the inside and cause vehicles to chew through turn signals bulbs also)
How expensive are these rapairs?
the daytime running light module is relatively easy to get to under the driver side kick panel
if you can replace the module yourself, the part can be gotten for around $130.00
is there something different that governs the rear blinkers? I'venever had to replace a bulb there!
its not a different circuit, i believe the rear bulbs are a higher amperage bulb and that is why they don't seem to be affected by the faulty components, the rear turn signals never seem to be affect, just the front, and usually the right side (passenger side) more than the left.
The right side was the one where we replace the whole headlight assembly, but it unfortunately went kaput too
I'm assuming the headlamp assembly failed in a different manner? neither of the components Ive recommended usually damage the entire headlamp, just the turn signal bulbs.
We just replaced it beause that was all we could think of to do!
on the plus side, im assuming that means you were having bigger problems with the right side than the left, which is usually the case when either the daytime running light module or the turn signal switch are at fault
So you would recommend trying the running light module?
last i checked the only place to get the daytime running light module was the dealer, and the turn signal switches were available from aftermarket suppliers. But the daytime running light module were still cheaper than the turn signal switches
if you have the time, i would advise to expose the module
check it for a burnt smell
and visually check it for burnt contacts
if is seems faulty or iffy, id replace is as a solid educated guess
Won't be me who does it....I consider it an accomplishment to change the bulbs, and my husband is even less mechanical than I am!
if it looks prefect.....then we have to choose between a cheaper repair and a more expensive one, without any real facts leaning us one way or the other
HA, congrats on being more mechanical than him
Almost sounds like new-car time......
its really not too bad, the regal is a strong car, but with known issues
Well, there are other problems - odometer display, gas gauge, etc. I have to use my gps as an odometer
the vehicle itself is extremely strong, it just happens to have a few issues that basically every one of them will encounter in its lifetime
mine has lived a longer life than most will ever see
Hmm... I'm not familiar with a recurrent issue with the instrument cluster on these vehicles.
maybe mine is the only one
all in all, id say a turn signal issue isnt worth getting rid of a car, but if you are having multiple other issues that are making you unhappy with it..
just remember, with a new car comes a car payment, and/or new problems ( only sometimes)
I bought a car with all the bells and whistles - my next one is going to be so basic, I'll have to hand crank the windows!
a new car or used?
no - the one I bought was new - my next one is going to be used
cutting down on the options can save alot of money on cost and on unkeep costs with both new and used
Well, I do thank you for your advice. This will give me something to try.
You are very welcome, take care. please don't forget to click accept if I have helped you.Let me know if there is any further information I can provide for you
Thanks again!


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My left wrist tattoo, the sun, is about two years old. My right wrist, the flowers, is only about a year old. Is there any chance to be able to have surgical excision?
Both my Wrists Are Tattooed, I Want a Surgical Excision, is There Help for Me? (photo)
Doctor Answers 5
Surgical Excision Versus Laser Tattoo Removal
Surgical excision would leave permanent scars. Also your tattoos are too large for this procedure. Laser tattoo removal would be your best choice. The black sun tattoo would be the easiest to remove since black ink absorbs all wavelengths of light. The black outline and stems of your tattoo would also be fairly easy.
The blue and the yellow would be a little more challenging.
Look into the 1064nm Q-switched Nd:YAG lasers ( eg Spectra and medlite lasers) for tattoo removal. This system can address multiple colors. And it is safe for all skin colors. Always consult your dermatologist for his/her advice.
Learn more: http://www.finetouchdermatology.com/los-angeles-laser-tattoo-removal/
Removing tattoos with surgical excision
is never a good idea unless they are very small in size or there is considerable excesses of skin to allow a surgical excision. l
Lasers are most effective with blues and blacks... you could have trouble completely clearing your yellow and perhaps may be better off not removing those and covering it with clothing instead.
Best of luck with clearing your skin.
Tattoo removal
Surgical excision is a poor choice, photo therapy is a better option for tattoo removal.
You might also like...
Surgical Excision of Tattoos?
Thank you for the question and pictures.
Unfortunately, you are not a candidate for any type of excisional procedure. Laser tattoo removal is your best option with the understanding of the limitations associated with this procedure. Multiple treatments will be necessary and some of the colors (especially the blue) will be very difficult to remove. There may always be telltale signs that tattoos were once present. These signs however will be much better than scars left after excision.
On the positive side, much of the ink is black ( favorable) and the tattoo is on your upper extremity ( favorable).
I would suggest an person consultation with a well experienced plastic surgeon or dermatologist.
Best wishes.
Learn more: http://www.poustiplasticsurgery.com/Procedures/procedure_lasertattooremoval.htm
Both my Wrists Are Tattooed, I Want a Surgical Excision, is There Help for Me?
When it comes to excising large pigmented lesions and tattoos, such large surface deformities cannot be removed at one time UNLESS you have excess skin which would permit it. Unfortunately, no one has that much excess skin of their distal exttremities.
Your BEST option is to have several laser treatment. You will be able to return to work in between sessions AND once the tattoos are broken up unlike a surgical removal you will not have extensive visible scarring which will always be raising questions and suspicions. If you insist on having them excision, they may be taken out in pieces (central portion out first and by re-excising the central portions several months later the whole tattoo can be taken out over several years). Another option is to put a balloon expander in and fill it up generating more skin. Once more skin has been produced, the tatoo is cut and the excess skin is used to close the opening.
Good Luck.
Peter A Aldea, MD
These answers are for educational purposes and should not be relied upon as a substitute for medical advice you may receive from your physician. If you have a medical emergency, please call 911. These answers do not constitute or initiate a patient/doctor relationship.


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If you look at the list of popular boys' names—and some girls' too—you start to feel that almost every conceivable surname has become a first. Occupational names like Mason and Sawyer, patronymics like Jackson and Addison, Irish surnames Nolan and Quinn, Old Hollywood glamour last names such as Harlow…the list goes on.
Of course the ideal scenario for coming up with a fresher choice would be to discover some surname surprise on your own family true, but that isn't always possible.
So are there any more original choices in this category of surname baby names still to be discovered? Of course there are…in all the above modes and beyond. Here are a few ideas to get you started; some have been lightly used over the years, but they all rate consideration for wider use.
BODIN/BOWDEN—Bodin is the Swedish form of Booth; Bowden is an English and Scottish place name–with either spelling, would fit right in with classmates like Holden and Colton.
BRANNIGAN—A possible update of Brandon, with the nutritious bran vibe plus a lively Irish lilt; there's also the starstruck BRANDO.
BRISCO/BRISCOE—An energetic English place name with the great o-ending. Lennie Briscoe was a long-running character on Law and Order played by Jerry Ohrbach. The Brisco spelling also relates to single-named American rapper Brisco, who was actually born with the first name British.
BROPHY—A hearty Irish surname that's much more distinctive than Brody or Murphy. And it has the pleasing rhyme with the word trophy.
CAFFERTY—Everybody loves Rafferty, so how about cousin Cafferty, the meaning of which relates to horses?
CALHOUN—There are several cool paths to the nickname Cal other than Calvin, including Calhoun, Calvino and Callahan. John Calhoun was a historic 19th century US political figure; Noah Calhoun the character played by Ryan Gosling in the 2004 film The Notebook, based on the Nicholas Sparks novel.
CHEEVER—A name with strong literary resonance via novelist and short story writer John, this cheery name also has subliminal ties to the desirable word 'achiever.'
CRUSOE—Conjures up the adventurous image of Robinson from the 1719 novel and the numerous films, TV shows and opera based on him. (And Robinson's pretty good too.)
DENNISON—A solid patronymic that could make an interest choice moving beyond the son of Dennis.
DUGAN—An open, friendly Irish surname with the cheery 'oo' sound.
DRYDEN—Strong, but also poetic, with its literary association to influential Restoration poet John.
ESSEX—The name of an English county and the Earl who had close ties to Queen Elizabeth I (portrayed in the film Elizabeth and Essex by the dashing Errol Flynn), it boasts the desirable 'x' factor.
FINNEY—Finn names are everywhere—Finn, Finlay/Finley, Finnian, Finnegan—and this one has a whimsical, nickname feel.
GRANGER—An occupational surname with an agricultural aura, much more cultivated than the rhyming Ranger.
HUTTON—Kind of a cross between Hudson and Sutton. Remember that old advertising slogan, "When E.F. Hutton talks, people listen." …? Also associated with heiress Barbara, hyperkinetic musical actress Betty and model Lauren.
IMBRY/IMBRY/EMBRY— All these names have a sweet feminine nickname feel. Liz Embry was the lady photographer in The Philadelphia Story.
KIMBER—Just drop the ly from Kimberly and you have a much stronger, more modern-sounding unisex name. Kimber is associated with a medieval convert to Christianity who founded a monastery and was venerated as a saint.
LORCA—Not completely undiscovered, this lovely Spanish name was used by singer/poet Leonard Cohen for his now grown daughter. Lorca is a place name in the Spanish province of Navarre, but more famous as the surname of the eminent poet and playwright Federico Garcia Lorca—the direct inspiration for the name of Lorca Cohen.
QUAID/QUADE—Quaid, as in the acting family of brothers, brings a perfectly usable first into the limited Q-field. The Aussies have been known to use the Quade spelling, inspired by rugby star Quade Cooper. Since it's a Gaelic surname meaning "son of Walter," there's also a subtle raison d'être right there.
TOLLIVER—This Scottish occupational surname—it was used for metal workers—makes for a lively extension of Oliver, with Tolly as a charming substitute for Ollie. Melba Tolliver is a barrier-breaking TV anchorwoman; Cy Tolliver was a character on Deadwood.
WHEELER—Wheeler definitely has the Whee! factor. It's an appealing er-ending name that has hardly been heard since 1909 and has some historical cred via onetime US veep William Almon Wheeler.
Any of these appeal? Do you have any undiscovered surnames you'd like to share?


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Rosebuds, beautiful range of erotic jewelry, made and designed in France by Julian Snelling.
This big model is designed for those already accustomed to anal pleasure.
The Rosebud can be used for numerous practices. Anal ornament or more active toy to increase your sensations and your pleasure, it is a discreet and elegant companion, sign of a certain erotic refinement. Whether you wear it during sexual games or during the day, it is invisible under your clothes. Very effective for sportsmen, especially while cycling, jogging and swimming, the simplest walk becomes a sensual feast. Its caress is renewed with each step... When you wear a Rosebud, the sitting position feels totally different. Inserted a few hours before a sexual encounter, it will also heighten pleasure during preliminaries.
Apart from providing voluptuous sensations, wearing a Rosebud for a prolonged time will also re-educate and stimulate the perineum whose tonicity is rather weak in the Western World, although it is a powerful and main actor in male and female orgasms and essential for vigorous erections and their control.
Material: Stainless steel
Colour: Clear Crystal
Size: 96 mm - Ø 40 mm, penetrating length 84 mm - Ø 40 mm
Weight: 440 g
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Hi there! I wanted to share my experience as this site has been so helpful for me. I am 33, mommy of 2 wonderful children 9 and 11. I am 5'4, 130lbs and this is my 3rd surgery. 2 years ago I had smart lipo of outer thighs at that time i had almost no cellulite. Over that past 2 years I became ridden with cellulite as it healed. today I had cellulaze outer and back thighs, and lipo of abdomen, inner and outer thigh. Counting on Dr. Greenberg to fix me!
Cellulaze/lipo Combo Mommy of 2 - Woodbury, NY
Hi there! I wanted to share my experience as this...
Today was the day, so I may be a lil loopy, lol. I would first like to say that results are the most important part of selecting a doctor. People on here talk about his bed side manner, the fact that you don't see him much, or that he doesn't take his time. In my experience and expectations thus far (just got home from procedure) he was sweet, calming, took time in consult and pre-op and always asked of I had anymore questions. His staff is wonderful and informative. It's not his job to hold your hand, it's results! And since I just got this done today, I will update on Dr G and my results soon.
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