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Pulmonary hypertension (PH) causes loss of body weight and inspiratory (diaphragm) muscle dysfunction. A model of PH induced by drug (monocrotaline, MCT) has been extensively used in mice to examine the etiology of PH. However, it is unclear if PH induced by MCT in mice reproduces the loss of body weight and diaphragm muscle dysfunction seen in patients. This is a pre-requisite for widespread use of mice to examine mechanisms of cachexia and diaphragm abnormalities in PH. Thus, we measured body and soleus muscle weight, food intake, and diaphragm contractile properties in mice after 6–8 weeks of saline (control) or MCT (600 mg/kg) injections. Body weight progressively decreased in PH mice, while food intake was similar in both groups. PH decreased (P<0.05) diaphragm maximal isometric specific force, maximal shortening velocity, and peak power. Protein carbonyls in whole-diaphragm lysates and the abundance of select myofibrillar proteins were unchanged by PH. Our findings show diaphragm isometric and isotonic contractile abnormalities in a murine model of PH induced by MCT. Overall, the murine model of PH elicited by MCT mimics loss of body weight and diaphragm muscle weakness reported in PH patients.
Citation: Ahn B, Empinado HM, Al-Rajhi M, Judge AR, Ferreira LF (2013) Diaphragm Atrophy and Contractile Dysfunction in a Murine Model of Pulmonary Hypertension. PLoS ONE 8(4): e62702. https://doi.org/10.1371/journal.pone.0062702
Editor: Rebecca Berdeaux, University of Texas Health Science Center at Houston, United States of America
Received: December 19, 2012; Accepted: March 25, 2013; Published: April 22, 2013
Copyright: © 2013 Ahn et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This study was supported, in part, by funds from National Institutes of Health grant R00-HL098453-01. No additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Pulmonary hypertension (PH) is a chronic, progressive disease characterized by inflammation and pulmonary vascular remodeling leading to high blood pressure in the pulmonary circulation [1]. Symptoms of PH include breathlessness, weakness, and fatigue. These symptoms have been traditionally attributed to pulmonary gas exchange abnormalities [2]. However, important factors in PH symptomatology that have emerged in recent years are cachexia and skeletal muscle weakness [3], [4]. Inspiratory muscle weakness has been documented in PH patients [5]. The diaphragm is the primary muscle used during inspiratory breathing efforts; hence, depression of diaphragm force is of particular relevance for symptoms of breathlessness and fatigue experienced by patients with PH. Depression of muscle force can result from atrophy (decrease in fiber size or cross-sectional area) and contractile dysfunction (defined as force normalized for muscle cross-sectional area). Patients with PH also experience loss of body weight [4], which is associated with poor prognosis in chronic diseases, e.g., heart failure [6]. However, diaphragm weakness and loss of body weight remain poorly understood in PH.
Diaphragm weakness in PH results from loss of muscle fiber size (atrophy) and contractile dysfunction [3], [7]. Contractile dysfunction has been shown by lower maximal diaphragm isometric tension normalized for cross sectional area in rats and humans with PH [7]–[9], whereas the effects of PH on diaphragm isometric force in a mouse model of PH remains unknown. In addition, impaired ventilatory function and diminished inspiratory pressure during dynamic maneuvers suggest diaphragm isotonic contractile impairments in patients with PH, but could also be due to abnormalities extrinsic to the diaphragm muscle (e.g., neurological or airway narrowing). However, it is unclear whether PH impairs diaphragm isotonic contractile properties.
Monocrotaline (MCT) has been widely used to induce PH and right ventricular (RV) failure in rodents [10]–[15]. To our knowledge, studies focusing on skeletal muscle abnormalities caused by PH and RV failure induced via MCT have been done solely in rats [8], [14], [16], [17]. Rats are hypersensitive to MCT – 10 times more sensitive than mice [12] – and become severely anorexic upon exposure to MCT [14]. While PH causes loss of body weight [4], it appears that PH per se does not necessarily cause anorexia in humans [18], [19]. Mice receiving MCT develop PH and show a progressive loss of body weight like rats [15], [20], but it is unknown if mice receiving MCT are anorexic. This aspect is relevant for further understanding diaphragm abnormalities in rodent models of PH.
The primary purpose of the present study was to investigate the effects of PH on diaphragm fiber atrophy and contractile properties in the mouse model of the disease induced by MCT. We found that PH caused diaphragm fiber atrophy and impaired isometric and isotonic contractile function in mice. As potential mechanisms for contractile dysfunction in chronic diseases, we tested for protein oxidation and selective degradation of myofibrillar proteins and found those variables were unaffected by the disease. A secondary purpose of our study was to examine changes in body weight and food intake in mice treated with MCT. We observed that progressive loss of body weight in PH mice occurred in the absence of anorexia.
Animals and procedures
We strictly followed all recommendations for the care and use of laboratory animals established by the National Institute of Health [21]. The study was also approved by the Institutional Animal Care and Use Committee at the University of Florida (protocol # 201105762), and all efforts were made to minimize animal suffering. We used C57BL6 male mice aged 8–12 wks old at the beginning of the experiments (Harlan, Indianapolis, IN).
Mice were individually caged, exposed to a 12∶12 hr dark:light cycle, and had access to standard water and chow ad libitum. We administered weekly injections of 600 mg/kg MCT (20 μL/g body weight, subcutaneous) or equivalent volume of sterile saline for 6–8 weeks to induce PH as shown by Yamazato et al. [15]. We measured body weight and food consumption 2–3× per week. On the day of the experiment, we anesthetized the mice with subcutaneous injection of ketamine (100 mg/kg) and xylazine (15 mg/kg). When the animals reached the surgical plane of anesthesia, we performed a laparotomy to excise diaphragm, heart, and lungs. One hemidiaphragm was quickly frozen in liquid nitrogen for biochemical analyses. The other hemidiaphragm was used to for histology and to measure contractile properties in vitro (see below). The heart was dissected to remove atria, major vessels, and separate the right ventricle (RV) from left ventricle plus septum (LV+S). Lungs, RV, and LV+S were blotted dry and weighed. We also removed soleus and blotted dry prior to measuring weight.
We used standard procedures for tissue preparation as outlined elsewhere [22], [23]. Briefly, diaphragm bundles were were embedded in Tissue-Tek freezing medium, frozen in liquid-nitrogen-cooled isopentane, and stored at −80°C. We sliced diaphragm bundles into 10 μm cross-sections using a Microm HM 550 cryostat (Microm International, Walldorf, Germany). Sections were incubated in 0.5% Triton X-100 solution for 5 minutes at room temperature, then washed in PBS for 5 minutes. Sections were subsequently incubated in anti-Laminin primary antibody (1∶200; Sigma-Aldrich) for 1 hour at room temperature in a humid chamber. After primary antibody incubation, sections were washed in PBS 3× for 5 minutes and incubated in anti-Rabbit rhodamine red secondary antibody (1∶100; Sigma-Aldrich) in blocking solution for 1 hour at room temperature in a humid chamber. Sections were then washed 3× for 5 min in PBS. We used a Zeiss Axio Observer-A1 microscope (Carl Zeiss Microscopy, Jena, Germany) and AxioCam MRm3 camera to capture images and NIH ImageJ software for image analysis. We traced the cell membrane envelope delineated by laminin to measure the area of all visible fibers from each diaphragm muscle section.
Isolated diaphragm contractile properties
We dissected a diaphragm strip along with rib and central tendon in bicarbonate-buffered solution (in mM: 137 NaCl, 5 KCl, 1 MgSO4, 1 NaH2PO4, 24 NaHCO3, 2 CaCl2) gassed with a mixture of 95% O2 and 5% CO2 at room temperature. We used 4.0 silk suture to tie the rib to a glass rod and to attach the central tendon to a Dual-Mode Muscle Lever System (300C-LR, Aurora Scientific Inc., Aurora, Canada). The diaphragm strip was placed in an organ bath containing bicarbonate-buffered solution. We adjusted bundle length to attain maximal twitch tension (optimal length, Lo), increased the temperature of the organ bath to 37°C, added D-tubocurarine (25 μM) to the solution, and after thermo-equilibration (20 min) started our force-frequency protocol. The isometric force-frequency protocol consisted of pulse frequencies of 1–300 Hz interspersed by 1 min intervals. The stimulation protocol consisted of supramaximal electrical current (600 mA) delivered through platinum electrodes using a biphasic high-power stimulator (701C, Aurora Scientific Inc.) with pulse duration of 0.25 ms and 300 ms train duration.
To determine the force-velocity relationship, we used afterloaded contractions employing a protocol similar to previous studies [24]–[26]. In this protocol, the bundle was maximally stimulated (300 Hz) to shorten against an external load corresponding to 2–80% Po above the resting tension. Each stimulus lasted for 200 ms and was delivered with 2 min intervals. We measured velocity at least 10 ms after the initial change in length in the linear portion of the tracing. We normalized shortening velocity per optimal length (Lo) and force per cross-sectional area (CSA, kN/m2). To estimate the bundle CSA, we divided diaphragm bundle weight (g) by bundle length (cm) multiplied by muscle specific density (1.056) [27]. The force-velocity curve was plotted and fitted to the Hill equation [28]. We determined maximal shortening velocity (Vmax) as the velocity at zero force in the force-velocity relationship. Power output for each isotonic contraction was also calculated as Force × Velocity and is given in kN/m2 × Lo/s.
Preparation of diaphragm tissue lysates
We homogenized diaphragm samples on ice using Kontes Duall Tissue Grinders in muscle extraction buffer (20 mM Hepes; pH 7.4, 2 mM EGTA, 1% Triton-X100, 50% Glycerol, 50 mM β-Glycerophosphate, 1× Protease Inhibitor Cocktail, 1× Phosphatase Inhibitor Cocktail (Thermoscientific)) [29]. Homogenates were rotated end over end for 1 hr at 4°C and then sonicated. The samples were then centrifuged at 15,000 g for 2 min at room temperature to pellet insoluble debris. We kept the supernatant and determined its protein content using the DC protein assay (Bio-Rad Laboratories).
We loaded 10–15 μg of protein into 4–15% polyacrylamide gradient gels (Bio-Rad Laboratories) and performed electrophoresis at 200 V for 50 mins at room temperature, and then transferred proteins to low fluorescence PVDF membranes (Immobilon-FL) at 100 mA overnight at 4°C. We blocked the membranes in either Odyssey Blocking Buffer (LI-COR) for 1 hr at room temperature, and probed with a sarcomeric actin specific antibody (JLA 20; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA) and a anti-troponin T1-T2-T3 antibody (TT-98; AbCam) diluted 1∶1000 and incubated overnight at 4°C, followed by secondary antibodies (IRDye, LI-COR) diluted 1∶10,000 and incubated for 45 minutes at room temperature. We quantified the bands using Odyssey Infrared Imaging system (LI-COR, Lincoln, NE). We stripped the membranes using 1× stripping buffer (LI-COR) for 20 minutes, re-probed with α-tubulin antibody (Developmental Studies Hybridoma Bank), and determined the abundance of α-tubulin, which was used for normalization purposes.
Protein Carbonyls Assay
We measured protein carbonyls in whole diaphragm homogenates using the Oxyblot kit according to manufacturer's instructions (Millipore Corporation). Briefly, we reacted 20 μg of protein with 2,4 dinitrophenylhydrazine (DNPH) and immunoblotted (see above) using anti-DNP primary antibody and anti-rabbit secondary antibody (IRDye 800CW). We scanned the membranes and quantified integrated intensity in each lane using the Odyssey Infrared Imaging system (LI-COR). We stripped the membranes using 1× stripping buffer (LI-COR) for 20 minutes, re-probed with α-tubulin antibody (Developmental Studies Hybridoma Bank), and determined the abundance of α-tubulin. Protein carbonyls were normalized for α-tubulin.
Myofibrillar protein isolation [30]
We homogenized diaphragm samples with Kontes Duall Tissue Grinders in Standard Relax Buffer with Triton X-100 (SRB-X100; 75 mM KCl, 10 mM imidazole, 2 mM MgCl2, 2 mM EDTA, 1 mM NaN3, 1% v/v Triton X-100), and then centrifuged for 1 minute at 20,000× g at 4°C. We washed the pellet once with SRB-X100 and once with SRB (75 mM KCl, 10 mM imidazole, 2 mM MgCl2, 2 mM EDTA, 1 mM NaN3) centrifuging at 20,000× g for 1 min at 4°C and discarding the supernatant each time. The pellet was then resuspended in 1∶20 (w/v) SDS-Page industrial buffer (8M urea, 2M thiourea, 0.05M Tris pH 6.8, 75 mM DTT, 3% SDS), vortexed for 1 h and centrifuged at 20,000 g for 10 min at room temperature. The supernatant containing the clarified soluble sample is the myofibrillar protein enriched fraction and was stored at −80°C. We determined protein concentrations of the myofibrillar protein enriched fraction using the RC DC Assay (Bio-Rad, Hercules, CA). We loaded 15 μg of protein into a 4–15% polyacrylamide gradient gel (Criterion precast gels; Bio-Rad, Hercules, CA), protein molecular weight standards (Precision Plus, Bio-Rad), and subjected to gel electrophoresis for 55 min at 200 V at room temperature. The gel was stained with Coomassie blue overnight (Thermo Fisher Scientific, Waltham, MA) and washed with dH2O three times. We quantified the optical density of individual bands and the whole lane (total protein) using the Odyssey Infrared Imaging system (LI-COR, Lincoln, NE) to determine the abundance of specific and total myofibrillar proteins. Specific proteins of interest were identified based on their known molecular weights, the band was quantified and data were normalized to total protein.
We administered MCT to a total of 14 mice. Of those, 2 animals died spontaneously and 2 were euthanized due to poor health conditions as recommended by institutional veterinarians. Thus, data from a total of 10 mice that survived the entire duration of the study (6–8 weeks) are reported here. In the cases of spontaneous death or euthanasia, the profile of changes in body weight resembled that of animals surviving the 6–8 weeks of treatment.
Several laboratories have used MCT to induce PH in mice [12], [13], [20]. The protocol we used has been shown to promote a 2.5 fold increase in right ventricular systolic pressure (RVSP) in C57BL6 mice over 8 weeks [13], [15], [20]. In this setting, PH causes RV hypertrophy defined by an increase in the ratio of RV weight to LV+S weight [RV/(LV+S)] of ∼20–25%. Due to technical problems, we were not able to measure RVSP, but MCT increased RV/(LV+S) by ∼18±2% in our study. We also observed other typical signs of PH after 6–8 weeks of MCT injections, such as increased RV and lung weight (Table 1). Hence, we consider that mice in the MCT group developed PH consistent with previous studies [13], [15], [20] and, heretofore, refer to this group as PH for clarity.
PH caused a progressive decrease in body weight after the first week of treatment (Figure 1). The decrease in body weight was accompanied by ∼15±1% lower soleus weight (Table 1). Importantly, food intake normalized for body weight was similar between groups and unchanged over the course of 8 weeks (Fig. 1C). The outcome was the same when absolute food intake was analyzed as average food intake during the 8-week period was 4.2±0.2 g/day for control and 4.3±0.4 g/day for PH.
A) Images of sample mice in each group at the end of study illustrating that PH mice are smaller than controls. B) Body weight expressed as percentage of value at the onset of study. C) Food intake expressed as mg per g body weight per day in each week of the study period. Data are from mice receiving control and monocrotaline injections for 8 weeks (N = 4 for Control; N = 5 for PH). * P<0.05 for control vs. PH.
We conducted morphological analysis in cross-sections of diaphragm bundles and measured diaphragm function in isolated bundles in vitro. Fiber cross-sectional area was decreased by 25% in PH mice (Fig. 2A–B). Due to problems in trials of RVSP measurements (see above), we were unable to determine diaphragm contractile properties in 4 saline-injected mice. Saline injections had no effect on diaphragm contractile properties when compared to data from age-matched mice. Thus, we included in our control group data of diaphragm function from age-matched mice that did not receive injections. PH decreased submaximal and maximal specific isometric tetanic force by 25–30% (Fig. 2C). Twitch force was unchanged (6.3±0.4 N/cm2 for control; 5.5±0.45 for PH) and, as a consequence, the twitch-to-tetanus ratio increased in PH. Time to peak tension (14±0.5 ms for control; 15±1 ms for PH) and one half relaxation time (17±1.1 ms for control; 17±1.4 ms for PH) did not differ between groups.
A) Cross sections taken from diaphragm muscle fiber bundles of sample control and PH mice. B) Diaphragm fiber cross sectional area of 200–300 fibers per muscle, from 3–4 mice per group. C) Specific Force, force normalized to bundle cross-sectional area. Data are from diaphragm bundles of controls and mice receiving monocrotaline for 6–8 weeks to induce PH (N = 10/group). * P<0.05 for PH vs. control.
An important and novel observation was that PH impaired isotonic contractile properties in the diaphragm. Specifically, maximal shortening velocity (Vmax) was 40% slower and peak power was 63% lower in PH than control (P<0.05; Fig. 3), despite a 138% increase in curvature of the force velocity relationship. The differences in the force-velocity characteristics remained after normalizing for Po (Fig. 3A).
A) Force-velocity relationship from sample control (open circles) and PH mice (closed circles). Force is relative to maximum isometric tetanic force (Fo). Shortening velocity is normalized to optimal bundle length for twitch force (L0). Solid and dashed lines are best fit from Hill equation. B) Maximal shortening velocity (Vmax) determined from extrapolation of shortening velocity to zero force using Hill equation (see Panel A for example). C) Force-power relationship of sample control and PH mice. Power is calculated as specific force (kN/m2) multiplied to shortening velocity (L0/s) shown in panel A. D) Peak Power from all mice in each group. Data are mean ± SE. Data in panels C and D are from n = 6/group. * P<0.05 vs. control.
Based on our findings of impaired isometric and isotonic diaphragm contractile function, we examined post-translational modifications of myofibrillar proteins as potential mechanisms of dysfunction. We measured total protein carbonyls as marker of oxidation and found no change in the diaphragm of PH mice (Fig. 4). Nor were there changes in the abundance of carbonyls in individual bands (data not shown).
These findings suggest that PH does not increase protein oxidation in mouse diaphragm. Total protein carbonyls are normalized to corresponding α-tubulin and expressed relative to the mean for the control group (n = 4/group). Image shows membranes probed for protein carbonyls and α-tubulin. Lanes are examples of control and PH diaphragm.
Protein content measured by the RC-DC assay in myofibrillar protein enriched fractions was similar in control (1.20±0.17 mg/ml and PH (1.10±0.38 mg/ml). Based on the existing literature, we focused our analysis of myofibrillar protein abundance on MHC, actin, and troponin T. We found that the optical density of the band corresponding to myosin heavy chain (MHC, ∼220 kDa) in myofibrillar protein enriched fractions was similar in PH and controls (Fig. 5A). We also found that sarcomeric actin (Fig. 5B) or troponin T (Fig. 5C) were unchanged in whole muscle homogenates (Western blots).
A) Images show protein bands corresponding to myosin heavy chain (∼220 kDa) in examples of control and PH mice. MHC data were normalized for total protein measured as optical density of entire lane. Molecular weights were determined from protein standards (not shown). B and C) Images are examples of lanes from membranes probed for sarcomeric actin (B) and troponin T (C) in whole-muscle homogenates. Optical density of actin and troponin T were normalized to α-tubulin in the respective membranes. All bar graphs show mean ± SE of results expressed relative to mean of control (N = 4/group).
The main findings of our study were that PH caused diaphragm atrophy and impaired both isometric and isotonic contractile properties in mice. Moreover, food consumption of PH mice was unchanged despite a progressive decrease in body weight over 6–8 weeks. These alterations in contractile function occurred in the absence of increases in protein oxidation or decreases in myofibrillar protein abundance.
Patients with PH experience loss of body weight, although the exact prevalence is unknown [4]. Patients with PH who experience loss of body weight have increased mortality rate after lung transplantation [31]. Loss of muscle mass is an important component of loss of body weight in chronic diseases [32]–[34]. Accordingly, soleus muscle mass of PH mice was 15% lower than control. The decreases in body and muscle weight were not caused by differences in food intake. In contrast, rats show ∼78% decrease in food intake 20 days after a single MCT injection compared to control [14]. This is an important observation when considering the experimental design of studies using MCT in rodents. To our knowledge, there are no published data showing that PH patients become anorexic, and it appears that PH per se does not cause anorexia in patients [18]. Our study shows that loss of body weight in PH mice is accompanied by decreases in muscle mass, and these changes are not caused by anorexia. Hence, our findings suggest that mice can be used to investigate the mechanisms of diaphragm contractile dysfunction in PH without confounding effects of anorexia induced by MCT.
Patients with PH have a 25–35% lower maximal inspiratory pressure than healthy peers [6], [35], which suggests diaphragm weakness. This weakness can arise from fiber atrophy and contractile dysfunction. Previous studies found diaphragm fiber atrophy in rats and patients [3]. Moreover, there was diminished specific force (force normalized for cross sectional area) in diaphragm bundles of rats with PH induced by MCT [3]. The decrease in specific force was evident in permeabilized single fibers of rats and patients with PH [3], [7], showing that PH causes dysfunction of the contractile apparatus.
In agreement with previous studies [3], [7], we found diaphragm fiber atrophy and decreased submaximal and maximal specific isometric tension in PH mice. The effects of PH on the diaphragm force-frequency relationship were slightly different in mice and rats. In mice, force was depressed predominantly at high stimulus frequencies (>50 Hz). In rats, the depression of force occurred at all frequencies [3]. These discrepancies might reflect the hypersensitivity of rats to MCT. The important point, however, is that mice injected with MCT to cause PH show diminished isometric specific tetanic force as seen in patients with PH. Specific force (reported here in kN/m2) represents force normalized for bundle cross sectional area, thus a decrease in specific force suggests contractile dysfunction independent of changes in muscle or fiber size. Thus, the ability of the diaphragm to generate force (or pressure) in PH is dimished due to combined effects of decreased fiber size (atrophy) and contractile dysfunction.
The force-frequency characteristics provide useful information regarding the force generating capacity of diaphragm, but existing data are limited to isometric contractions. The diaphragm rarely performs isometric contractions in vivo [25]. Rather, it shortens against submaximal loads. Therefore, shortening velocity and power output are more physiologically relevant properties of diaphragm function. Patients with PH show a 20% decrease in maximal sniff nasal pressure [6] – a volitional dynamic maneuver that involves diaphragm shortening. Similarly, maximal voluntary ventilation is 30% lower in patients with primary PH than in healthy controls [2]. The decrease in sniff nasal pressure and maximal voluntary ventilation in PH patients suggest, among other factors, impairment in isotonic contractile properties of the diaphragm. Indeed, we found 40% slower Vmax and 60% lower peak power in the diaphragm of mice with PH. There was also a downward shift in the force-velocity relationship when submaximal loads were normalized for maximal tetanic force, e.g., Fig. 3A. Because we used afterloaded isotonic contractions, the downward shift reflects slower cross-bridge kinetics due to factors intrinsic to cross-bridge cycling or the level of calcium activation [24]. Importantly, afterloaded isotonic tests simulate the contractile profile in vivo, and our data supports the notion that PH impairs the ability of the diaphragm to sustain ventilatory tasks. Impairment of isotonic function could, then, contribute to dyspnea in PH. However, the mechanisms underlying contractile dysfunction are unclear.
Oxidants decrease calcium release and calcium sensitivity of the contractile apparatus [36], [37], and impair sarcomeric protein function [38]. However, PH did not change protein oxidation status measured by abundance of carbonyls in whole-diaphragm homogenates. Our results corroborate the data from a recent study showing unchanged protein carbonyls in PH rats [9]. Administration of MCT in rats increases protein carbonyls in limb muscles [16]. Thus, protein oxidation may be specific of limb muscles or become apparent after the development of right ventricular failure. Additionally, we cannot discard the possibility that PH increases other oxidant-mediated modifications (e.g., thiol oxidation). Based on the current and recent data [16], we consider that PH-induced diaphragm isometric and isotonic contractile dysfunctions are not caused by increased protein oxidation.
A mechanism that has been proposed to explain diaphragm weakness in chronic diseases is selective degradation of myofibrillar proteins [39]–[41]. A recent study has shown that MHC abundance was unchanged in rats with PH induced by MCT [16], which is consistent with our data in mice. Loss of thin-filament proteins could also contribute to contractile dysfunction. PH elevates systemic levels of pro-inflammatory cytokines [17]. The pro-inflammatory cytokine interleukin 1 promotes degradation of sarcomeric actin [42], which would cause contractile dysfunction. Tumor necrosis factor-α stimulates selective degradation of troponin T that results in diminished isometric force [43]. We measured the abundance of actin and troponin T and found that both were unchanged in MCT mice. Thus, our findings substantiate the concept that selective degradation of myofibrillar proteins does not appear cause contractile dysfunction in PH [7]. Rather, post-translational modifications are the likely cause of isometric and isotonic contractile impairments in PH. Identification of these post-translational modifications was beyond the scope of our investigation, but PH may affect the phosphorylation or thiol oxidation status of myofibrillar proteins [44]–[46].
Our conclusion is that MCT-induced PH causes diaphragm atrophy and impairs diaphragm isometric and isotonic contractile properties in mice. These perturbations in isotonic diaphragm contractile properties likely play a role in impaired ventilatory responses of PH patients [47]. Contractile dysfunction in PH mice occurs in the absence of selective degradation of myofibrillar proteins or accumulation of protein carbonyls, which suggests post-translational modifications of myofibrillar proteins such as phosphorylation and thiol oxidation as alternative mechanisms for contractile impairments. Importantly, MCT causes loss of body weight but not anorexia.
Hence, our findings suggest that mice can be used to investigate the mechanisms of diaphragm muscle weakness and atrophy in PH without confounding effects of anorexia induced by MCT.
Author Contributions
Conceived and designed the experiments: BA LFF. Performed the experiments: BA HME MA LFF. Analyzed the data: BA HME MA ARJ LFF. Contributed reagents/materials/analysis tools: ARJ LFF. Wrote the paper: BA HME ARJ LFF.
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- 31. Habedank D, Ewert R, Hetzer R, Anker SD (2009) Reversibility of cachexia after bilateral lung transplantation. Int J Cardiol 133: 46–50.
- 32. Hasselgren PO, Fischer JE (2001) Muscle cachexia: current concepts of intracellular mechanisms and molecular regulation. Ann Surg 233: 9–17.
- 33. Tisdale MJ (2002) Cachexia in cancer patients. Nat Rev Cancer 2: 862–871.
- 34. Fearon KC, Glass DJ, Guttridge DC (2012) Cancer cachexia: mediators, signaling, and metabolic pathways. Cell Metab 16: 153–166.
- 35. Meyer FJ, Lossnitzer D, Kristen AV, Schoene AM, Kubler W, et al. (2005) Respiratory muscle dysfunction in idiopathic pulmonary arterial hypertension. Eur Respir J 25: 125–130.
- 36. Andrade FH, Reid MB, Allen DG, Westerblad H (1998) Effect of hydrogen peroxide and dithiothreitol on contractile function of single skeletal muscle fibres from the mouse. J Physiol 509 (Pt 2): 565–575.
- 37. Moopanar TR, Allen DG (2005) Reactive oxygen species reduce myofibrillar Ca2+ sensitivity in fatiguing mouse skeletal muscle at 37 degrees C. J Physiol. 564: 189–199.
- 38. Callahan LA, She ZW, Nosek TM (2001) Superoxide, hydroxyl radical, and hydrogen peroxide effects on single-diaphragm fiber contractile apparatus. J Appl Physiol 90: 45–54.
- 39. Argadine HM, Hellyer NJ, Mantilla CB, Zhan WZ, Sieck GC (2009) The effect of denervation on protein synthesis and degradation in adult rat diaphragm muscle. J Appl Physiol 107: 438–444.
- 40. van Hees HW, Li YP, Ottenheijm CA, Jin B, Pigmans CJ, et al. (2008) Proteasome inhibition improves diaphragm function in congestive heart failure rats. Am J Physiol Lung Cell Mol Physiol 294: L1260–1268.
- 41. Ottenheijm CA, Heunks LM, Dekhuijzen RP (2008) Diaphragm adaptations in patients with COPD. Respir Res 9: 12.
- 42. Li W, Moylan JS, Chambers MA, Smith J, Reid MB (2009) Interleukin-1 stimulates catabolism in C2C12 myotubes. Am J Physiol Cell Physiol 297: C706–714.
- 43. Adams V, Mangner N, Gasch A, Krohne C, Gielen S, et al. (2008) Induction of MuRF1 is essential for TNF-alpha-induced loss of muscle function in mice. J Mol Biol 384: 48–59.
- 44. Solaro RJ, Kobayashi T (2011) Protein phosphorylation and signal transduction in cardiac thin filaments. J Biol Chem 286: 9935–9940.
- 45. Canton M, Neverova I, Menabo R, Van Eyk J, Di Lisa F (2004) Evidence of myofibrillar protein oxidation induced by postischemic reperfusion in isolated rat hearts. Am J Physiol Heart Circ Physiol 286: H870–877.
- 46. Canton M, Skyschally A, Menabo R, Boengler K, Gres P, et al. (2006) Oxidative modification of tropomyosin and myocardial dysfunction following coronary microembolization. Eur Heart J 27: 875–881.
- 47. Schwaiblmair M, Faul C, von Scheidt W, Berghaus TM (2012) Ventilatory efficiency testing as prognostic value in patients with pulmonary hypertension. BMC Pulm Med 12: 23.


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Lowell Tuplin demonstrates how he treats his elm tree to help slow the spread of the dreaded Dutch Elm Disease. He fills the drill holes with pure turpentine and then plugs the holes with earplugs.
Eric McCarthy/Journal Pioneer
The City of Charlottetown was busy fighting Dutch elm disease in 2013.
Coun. Rob Lantz, vice-chairman of the urban beautification, says the city removed 68 trees last year that were devastated by the disease but more than made up for it by planting 85 new trees.
"It is striking to see them come down. They (stumps) create a visual blight on the streetscape. We can't plant trees in the exact same place (but) there were more trees planted than were cut down.''
The issue came up when Coun. Mitchell Tweel noted the amount of tree stumps that have been removed and asked Lantz what the city's plan was.
Lantz estimates there is a net gain of 15 to 20 elm trees.
Dutch elm disease is a fungi spread by the elm bark beetle. It has devastated native populations of elms which had not had the opportunity to evolve resistance to the disease. It can affect any elm tree.
Since its introduction from Europe in 1930, it has destroyed millions of elm trees across North America.
The early symptoms appear from the latter half of June to the middle of July when the leaves on one or more branches may wilt, droop and curl. The leaves then turn brown and usually remain on the tree. If the tree is infected later in the summer, the leaves will droop, turn yellow and drop prematurely.
It should be noted that while diseased elm trees have been taken down and removed on private property, the city doesn't have the authority to replant trees on private property.
Due to the interest from various councillors on the issue at Monday night's public meeting, Lantz asked all of his colleagues to submit a list to his committee of where they would like to see elm trees planted in the city.

subset: commoncrawl


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Rebel Lens Photographer
_Kitty_Model Female Savannah, Georgia, US
My Website: Deviant ArtMy MM URL: http://www.modelmayhem.com/artzekitty
Mayhem # 2615634
About MeI took classes on how to model at the All American Modeling Agency until I turned 12. I did a few runway shows, was a window model for JcPenny, and then I stopped modeling for a while and focused on photography and school. I have an account on here mainly for my photography, but I am interested in getting back into modeling. I enjoy foot modeling the most, because who doesn't love wearing a pair of sexy shoes?! Even modeling barefoot can feel sexy.
P.S. - I ALWAYS bring someone with me to photoshoots if I haven't met you/ worked with you before.
Have you worked with _Kitty_? Add Credits for _Kitty_! >>
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Ports of Call
View all the ports that Seabourn can bring you to.
All Ports
Porto Belo, Brazil
Porto Belo attracts Brazilians and visitors to enjoy its watersports and beautiful beaches, either on the mainland or the small island just offshore. Venture inland to Florianopolis, to see colorful colonial period architecture and culture, or hop to Blumenau, a city on the Itajai-Acu River built by Germans in the 19th century and still dotted with strikingly Bavarian buildings. Its Oktoberfest is said to be one of the best in the world.


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Connect to Databases
porky allows you to connect to databases like MySQL and SQLite. Simply write SQL Statements and work directly with the results. It is possible to implement various other database drivers, too!
porky is a JavaScript extension for Adobe® InDesign® and other Adobe® products supporting ExtendScript
Use various data types like SQL databases, XML- and JSON files as a variable source.
Handle recurring tasks by automation – code in JavaScript.
Create entire workflows based on your JavaScript code.
porky integrates seamlessly into Adobe® InDesign®


text: I-25 Webcams: Fort Collins to Cheyenne By Townsquare Media October 26, 2012 6:28 PM | K99 Share on Facebook Share on Twitter Crossroads Blvd. Webcam (South of Fort Collins) Wellington Webcam Natural Forest Webcam (Near the Wyoming/Colorado Border) WYO Hill Webcam (South of Cheyenne) College Drive Webcam (Cheyenne) Source: I-25 Webcams: Fort Collins to Cheyenne Category: Colorado News Share on Facebook Share on Twitter More Leave a comment Send to a friend! Print this page Pin it! Reddit This! Share on Tumblr! Recommended For You ZergNet Best of the Web ZergNet


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Dec. 4, 2002
CONTACT: Ron Podell
YPSILANTI - Approximately 100 Eastern Michigan University students are helping lay the groundwork for a "wonderful life" for children in the pediatric emergency care unit at St. Joseph Mercy Hospital of Ann Arbor. EMU students are constructing scenery reminiscent of the 1945 holiday classic, "It's A Wonderful Life," which is being used as the theme for this year's St. Joseph Mercy Hospital Holiday Ball. The 27th Annual Holiday Ball is scheduled Saturday, Dec. 7, 6 p.m.-midnight, at EMU's Convocation Center, 799 N. Hewitt Drive.
All monies raised will be donated to the hospital's pediatric emergency/urgent care unit, according to Kelly Timmons, special events manager for St. Joseph Mercy Hospital.
Communication and theatre art students will design and construct scenery to mirror the town of Bedford Falls, the backdrop for "It's A Wonderful Life." The Encore Theatre Troupe will act as strolling carolers and other students will play characters from the movie and act out scenes from the film. A portion of the EMU Marching Band also will perform.
EMU ROTC students will greet the ball's guests while EMU ambassadors will handle hospitality duties.
Jan Brandon, a member of the EMU Board of Regents, is chair of the ball. Pam Kirkpatrick, wife of EMU President Samuel A. Kirkpatrick, is the entertainment committee chair.
For more information, call 487-4400.


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Madison Scholarly News — April 2006
Dr. Lennis G. Echterling (Professor, Graduate Psychology; Director, Counseling Psychology) was appointed to the Terrorism and Disaster Behavioral Health Advisory Council for Virginia, which was formed by the governor's office to provide guidance to Virginia's key disaster-response entities. The council's membership includes disaster experts from state agencies, community services boards, American Red Cross, military and academic institutions.
Dr. JoAnne M. Holman (Associate Professor, Media Arts and Design) was invited to join the Consumer Affairs and Outreach Working Group of the Consumer Advisory Committee, Federal Communications Commission. The committee is to make recommendations to the FCC regarding consumer issues within the jurisdiction of the commission and to facilitate the participation of consumers in proceedings before the commission.
Three faculty from the College of Arts and Letters received the 2006 Edna T. Shaeffer Humanist Award for summer research and writing on scholarly research project in the humanities, announced March 28: Dr. Liam Buckley (Assistant Professor, Anthropology) will research his project, "Is There a Home Away from Home? West African Portraiture and Transnational Aesthetics"; Dr. Maged Mikhail (Assistant Professor, History) will research "Egypt from Late Antiquity to Early Islam"; Dr. Debali Mookerjea-Leonard (Assistant Professor, English) will research "The Paradox of Independence: Literature and the Trauma of Partition" referring to the division of British India into India and Pakistan that accompanied decolonization in 1947. The award of $4,000 each is funded through a bequest of the late Evelyn Pugh, a 1936 Madison graduate and a career English teacher and is named in honor of Shaeffer, a music faculty member from 1915 to 1956.
Dr. Joann H. Grayson (Professor, Psychology) was selected to receive the 2006 Champion for Children Award by Prevent Child Abuse Virginia, which recognizes a long-term commitment to and investment in Virginia's children and their families by individuals and organizations.
Dr. Steve Stewart (Professor, Health Sciences) was selected to receive the American Association of Health Education's 2006 Professional Service to the Health Education Award for 2006, presented at the AAHE/American Association of Health, Physical Education, Recreation and Dance national convention, April 28. The award honors an individual who has demonstrated substantial service to AAHE as an officer, committee member, program participant and leadership to the health education profession.
Mark D. Rooker (Assistant Professor, Art and Art History) is exhibiting his artwork that tells science-fiction stories, "Mark Rooker: Jewelry," at Pittsburg State University in Kansas through April 5. Rooker also designed the exhibition's interactive display in PSU's University Gallery.
Melinda S. Burchard (Learning Strategies Coordinator, Disability Services) received $300 from Phi Delta Kappa to assist students with disabilities in the development of independence, responsibility and effective self-advocacy.
Sgt. Margaret A. Campbell (Crime Prevention Coordinator, Police and Safety) received $6,000 from CINTAS Corp. to obtain Automated External Defibrillators for the JMU police department.
C. Scott Coverstone (Support Services Division Commander, Police and Safety) received $95,248 from the Virginia Department of Emergency Management to provide equipment, training, planning and exercises for first responders to develop better preparedness to prevent, respond and recover from potential acts of terrorism.
Dr. Dorothy J. Della Noce (Assistant Professor, Communication Studies) received $8,056 from the Institute for the Study of Conflict Transformation Inc. to direct and execute the institute's knowledge creation and dissemination function.
Dr. Daniel M. Downey (Professor, Chemistry) received $8,000 from the U.S. Department of Agriculture Forest Service to inventory water quality of forest streams and to monitor the effects on stream water chemistry.
Dr. Steven W. Evans (Director, Alvin V. Baird Learning Disabilities Center; Professor, Psychology) received $1,500 from Eli Lilly & Co. to sponsor an attention deficit/hyperactivity disorder conference.
Dr. Joanne V. Gabbin (Executive Director, Furious Flower Poetry Center; Professor, English) received $20,000 from the R.R. Donnelley Foundation to promote poetry while assisting trauma survivors of Hurricane Katrina through producing an anthology of poetry written by hurricane survivors, observers and professional poets.
Elizabeth Haworth (Assistant Director, Furious Flower Poetry Center) received $500 from the Arts Council of the Valley for "Nikki Giovanni Presents 'Truth Is On Its Way': The 35th Anniversary Celebration Concert."
Dr. W. Christopher Hughes (Assistant Professor, Physics) and Dr. Gerald R. Taylor (Director Emeritus, Center for Materials Science; Professor Emeritus, Physics and Integrated Science and Technology) received $67,000 from the National Science Foundation to provide research experiences for undergraduates involving faculty from five academic departments and to focus on providing high-quality research experiences in materials science.
Henry A. Reeves (Director, Small Business Development Center) received $380,875 from the U.S. Small Business Administration to provide technical advice and guidance and to serve as a business and economic information focal point for small businesses.
Dr. Rosa E. Rivera-Hainaj (Assistant Professor, Chemistry) received $174,775 from the National Science Foundation to attract more students from under-represented backgrounds to obtain undergraduate degrees in the biological and chemical sciences.
Dr. Joseph D. Rumbo (Assistant Professor, Sociology) received a Faculty Initiative Development Grant from the Office of International Programs to present two papers at two conferences: "Exploring the Symbolic Universe of the Rainbow: A Framework for Doing Consumer Research at the Market's Edge," at the Annual Asia-Pacific Region Conference of the Association for Consumer Research in June 2006 in Sydney, Australia, and "Cultivating Alternative Self-Schemas and Lifestyles: Resisting the Normativity of Consumerism," to be presented at the International Centre for Anti-Consumption Research, also in June 2006, in Auckland, New Zealand.
Dr. Lynne F. Stover (Teacher Consultant, Center for Economic Education) received $1,647 from the National Council on Economic Education to conduct a teacher-training workshop on the Mathematics and Economics: Connections for Life, 3-5 curriculum series.
Dr. Grace A. Wyngaard (Professor, Biology) gave a presentation on reforms of the biology department's curriculum at the Biology Leadership Conference: Shaping the Future of Introductory Biology for Majors III, held in Bermuda.
Dr. Helmut Kraenzle (Professor, Integrated Science and Technology; Professor and Director, Master's Program in Remote Sensing and GIS at the United Arab Emirates University) presented the keynote address, "Tracking Shipping Containers with a Geospatial Information System for National Security," on the National Security Track of the Map Middle East 2006 Conference on Geospatial, Technology and Application, held in Dubai, United Arab Emirates, March 26-29.
Dr. Laura Lewis (Associate Professor, Anthropology) presented "Performing La América in Black-Indian Mexico: Race, Gender and the Nation," on the panel Place, Space and Stage: Performing Gender in the Americas at the XXVI International Congress of the Latin American Studies Association, San Juan, Puerto Rico, March. She also chaired the panel.
Dr. Laura Lewis (Associate Professor, Anthropology) presented her current research at the Afro-Mexican Studies Symposium at the Mexican Fine Arts Center Museum, Chicago, March 31-April 1.
Dr. Stephen Poulson (Assistant Professor, Sociology) presented "Political Opportunities, Legitimacy and the Strategies of Violence among Movement Groups in Iraq," at the Southern Sociological Society Annual Meeting, New Orleans, March 24.
Dr. Christine M. Robinson (Assistant Professor, Sociology and Interdisciplinary Liberal Studies) presented "The Social Structure of the Ex-Gay Movement in Virginia" at the Virginia Social Science Association Meeting in Richmond, March 25.
Dr. J. Peter Pham (Assistant Professor, Justice Studies; Director, Nelson Institute for International and Public Affairs) wrote "Strategic Horizons," a review essay published in the Spring 2006 issue of The National Interest, no. 83. The essay in the foreign policy journal focuses on four books published in 2005: "American Global Strategy and the 'War on Terrorism'" by Hall Gardner, "The American Era: Power and Strategy for the 21st Century" by Robert J. Lieber, "New Glory: Expanding America's Global Supremacy" by Ralph Peters and "Taming American Power: The Global Response to U.S. Primacy" by Stephen M. Walt. Pham also wrote "Torture, Democracy and the War on Terrorism," an essay published in vol. 6, no. 4 of The Long Term View, a journal of the Massachusetts School of Law at Andover. Pham presented the essay at a symposium, sponsored by the law school, which focused on the question "Are Our Highest Officials Guilty of Torture?"
Dr. Stephen Poulson (Assistant Professor, Sociology) published "Iranian Public Opinion Matters," in the "Roanoke Times," March 19.
Dr. Grace A. Wyngaard (Professor, Biology) is one of four authors of "Genetic architecture of the cryptic species complex of Acanthocyclops vernalis (Crustacea: Copepoda) II. Crossbreeding experiments, cytogenetics and a model of chromosomal evolution," an article published in the February issue of Evolution. (Click here for a NSF brief on the article.)
Sang Yoon (Professor, Art and Art History) had two logo designs, one for the Furious Flower Poetry Center at JMU and the other for Rich Cleaners, selected for publication in "American Corporate Identity 22." Harper Collins Publishers will release the full-color hardback book in late 2006. Yoon will receive a Certificate of Design Excellence for her work.


text:
From The Atlantic
A ten-day festival for purity and happy marriages.
Performers struggle to make ends meet.
During the "Bol Bom" pilgrimage, the faithful chant the name of Lord Shiva while running some 9 miles.
Water to cure what ails you.
A terror trail ends here, and anti-Monsanto activists clash with rural farmers


text:
2 days ago i did a 50% TCA peel on my stomach stretch marks. I have done peels in the past with great results. Right now the majority of my stomach is dry, cracked and peeling looking normal but there is a darker red patch that is not doing anything. It is not dry, cracked... It is smooth like my original skin just a bit red and sore. Pictures are more red than they appear in real life. I am afraid i have scarred my skin. I am not doing anything besides washing with soap and water. No creams.
TCA Chemical Peel 2 Days Ago On Stomach For Stretch Marks? (photo)
Doctor Answers 2
TCA Chemical Peel 2 Days Ago On Stomach For Stretch Marks?
Not sure if this was done by a doctor or did you self treat? Either way allow more healing time. If done by a doc than return to see him/her for guidance.
Do Not Use TCA on the Abdomen
Using TCA on the abdominal area not a good idea. Was this done by a physician, spa clinic or did you order it on the internet? Serious scarring may result from the ill advised use of this acid. Never use TCA on the body and it should only be applied by an experienced physician on the face. I would not expect any type of chemical peel to make stretch marks better. At this point you need to see a dermatologist or plastic surgeon and be started on a steroid cream.
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.


text:
J-SOx compliance
J-SOx is the short name for the Japanese version of the Sarbanes-Oxley law or SOx. SOx refers to a piece of United States legislation drafted by American politicians Sarbanes and Oxley. The law is intended to create transparency and reassure investors in the aftermath of a number of accounting scandals in the business world.
Since 2008, all companies that are active on the Japanese stock market have to comply with the Japanese version of this law, the Financial Instruments and Exchange Law. Daikin Europe N.V. has been J-SOx compliant since April 2008.
Daikin response to J-Sox
In response to the Japanese legislation, Daikin has realigned its systems to ensure the effectiveness of all operational processes throughout the Daikin Group that could affect financial reporting.
Daikin continually evaluates these systems to ensure the Company effectively provides the internal control assurance required by J-SOx. The company is committed to making any necessary adjustments to these systems, to ensure continued compliance with J-SOx and any related regulations.


text: Healthy Fast Food and Takeout Facebook Pinterest Twitter Print Mail Save Left Arrow Right Arrow Photo by Keate Left Arrow Right Arrow 12 of 13 Grid View See All Subway If You Usually Order:A classic tuna sandwich with Cheddar cheese and potato chips. Make It Healthier:Get a wheat roll, and add extra vegetables. If you can't omit the cheese, know that American cheese is the lowest-fat option, followed by provolone, with Swiss and Cheddar tied for last. Choose baked chips. Better Yet:Subway's tuna salad contains lots of mayonnaise, so a less cholesterol-raising option would be the Veggie Delite or Turkey-Breast Sandwich from the "6 Grams of Fat or Less" side of the menu. Start


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DETROIT (AP) — General Motors Co. says its Chevrolet Volt electric car will cost $41,000 when it goes on sale in November. The price is about $8,000 more than its closest rival, the all-electric Nissan Leaf.
GM will also offer a $350-per-month lease deal that's about equal to the Leaf's.
Both cars are eligible for a federal tax credit that will cut their prices by $7,500.
The Volt runs on battery power but has a small gas engine to generate electricity once the battery runs down. The 4-door sedan can drive up to 40 miles on a fully charged battery. The Leaf can go up to 100 miles on a charge but doesn't have a gas engine. It goes on sale in December.
GM says the Volt is a better value because it eliminates worries about running out of power.


text:
Boars, Gore, and Swords is hosted by stand-up comedians Ivan Hernandez and Red Scott. In each episode they break down HBO's Game of Thrones and George R.R. Martin's A Song of Ice and Fire. They also talk about movies, TV, science fiction, fantasy, and lots of other things. NSFW.
In this episode, Red and Ivan discuss the Davos V & Bran IV chapters of George R.R. Martin's A Storm of Swords (Catch up on past podcast episodes here to listen to previous chapter breakdowns). They also talk about Railroad speaking directly to listeners, Stannis's Bannermen vs. Ivan's twitter followers, the Nightfort, prince stew, Railroad writing Hodor high, and a TALKING TREE.
GET BGaS: RSS | On iTunes | Download episode


text:
- About
- Programs
- Resources
- Briefing Room
- Contact
U.S. Department of Transportation
Federal Highway Administration
1200 New Jersey Avenue, SE
Washington, DC 20590
FHWA Order M3000.1C
|FHWA PERSONNEL MANAGEMENT MANUAL Transmittal 18
|Classification Code
|Office of Primary Interest
|October 18, 2004
What is the purpose of this transmittal? This transmittal revises Part 1, Chapter 6, Section 3 (Authorizing Travel Expenses for Preemployment Interviews; Travel and Transportation to First Post of Duty) of the Federal Highway Administration (FHWA) Personnel Management Manual (PMM).
What are the major changes? No major changes have been made to this section. This section has been revised to update the titles of those who may authorize travel expenses for preemployment interviews and travel and transportation to the first post of duty. In addition, the section presents the information in plain language.
Michael J. Vecchietti
Associate Administrator of Administration


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Cattle Decapitation
We are stoked to announce this year's line-up for the return of the Hell On Earth Tour. Please welcome Cattle Decapitation, Broken Hope, Hideous Divinity and Gloryhole Guillotine!Read More
This Friday, on July 14th, Norwegian Death Metallers Execration will release their new album 'Return to the Void' via Metal Blade Records. Please surf over to youtube.com to check out 'Hammer of Vulcan'!Read More
Amon Amarth
Metal Blade to re-issue the next two Amon Amarth albums 'Fate Of Norns' and 'With Oden On Our Side' on vinyl as part of their Originals-series! Both out July, 28th!Read More
Metal Blade Records
Metal Blade Records and BMG are proud to announce the upcoming release of For the Sake of Heaviness: the History of Metal Blade Records on August 29th. Written by label founder and CEO Brian Slagel with Mark Eglinton.Read More
New Releases
Upcoming Releases
Latest News
Vesania premieres new video for "Notion" via NoCleanSinging.com!
- 16/12/15 – Poland's Vesania has launched a new video for...Read More
Fates Warning teams up with Sheet Happens Publishing to offer fans guitar transcription of classic album, 'A Pleasant Shade Of Gray'
- 14/12/15 – Progressive metal legends Fates Warning recently released...Read More
Cattle Decapitation and Abiotic to come to Europe with Suffocation for the "The Tyrants Of Death" tour in 2016!
- 14/12/15 – CATTLE DECAPITAITON will join SUFFOCATION on their...Read More
Allegaeon stellen neuen Sänger Riley McShanevor
- 14/12/15 – Band set to record new album this winter Allegaeon just...Read More
Pilgrim premieres new video for "Paladin" via MetalInjection.net
- 10/12/15 – Rhode Island's doom metal trio Pilgrim has launched...Read More
EXUMER launch landing page for "The Raging Tides"!
- 09/12/15 – 1st song, artwork and pre-order live! "The Raging...Read More
BRIMSTONE COVEN reveals details for new album, 'Black Magic'
- 08/12/15 – Launches video for title track On January 29th, West...Read More
KETZER releases studio trailer in support of new album "Starless"!
- 08/12/15 – German KETZER will release their brand new album...Read More


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Local hotel owners featured in Best Western commercials
Local hotel owners Bob and Anita Dir are featured in Best Western's new "Stay With People Who Care" ad campaign.
The Dirs own and operate the Best Western Plus Milwaukee Airport Hotel & Conference Center, 5105 S. Howell Ave. They were chosen from over 2,200 Best Western hotel owners and operators across North America.
The ads can be viewed here.
Post a comment / write a review.
Facebook Comments
Disclaimer: Please note that Facebook comments are posted through Facebook and cannot be approved, edited or declined by OnMilwaukee.com. The opinions expressed in Facebook comments do not necessarily reflect those of OnMilwaukee.com or its staff.


text:
Some sites may try to open pop-ups, which are new browser windows that may be hidden under the current window or displayed on the top.
Generally, pop-ups are used for advertising, but some sites may use them to show information without disrupting the currently open page.
Whenever Mozilla Firefox blocks a pop-up window, a message like this one will appear.
1) Click Options.
In this menu, you can choose to allow pop-ups for this website, open the pop-up blocker options menu or never show this message when pop-ups are blocked.
Let's see how to add a website manually.
2) Open the menu.
3) Go to Options.
4) Click Options.
5) Go to the Content tab.
If you want to allow all pop-up windows, deselect this box.
6) Click Exceptions.
7) Type the address of the web site for which you want to allow pop-ups and click Allow.
8) Click Close.
That's it! Now you know how to manage your pop-up blocker in Mozilla Firefox.
You must be logged in to post a comment.
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text:
St. Agnes of Bohemia Catholic School is a Pre-K through 8th grade preparatory school in the heart of Chicago's Little Village. For over 100 years, St. Agnes has dedicated itself to educating immigrant families for lives of faith, service and leadership. We encourage spiritual, intellectual, physical, emotional and social growth in the Catholic Tradition. We maintain high expectations for all members of our community to develop a sense of personal responsibility and self-motivation. This is accomplished within a Gospel-centered framework of justice and love. We do this so our students can be successful Christians and achieve excellence in high school, college and in life.
HOW STUDENTS LEARN: The student's ability to learn is strongly dependent upon a positive atmosphere and well-planned instruction.The classroom should inspire students to achieve their fullest potential, especially through cooperative learning projects. Instruction needs to address the learning styles of all students and be as individualized as possible. Learning should be experiential, improve critical thinking skills and instill a life-long desire to learn in each student.
HOME-SCHOOL CONNECTION: The student's first teachers are in the home, therefore, good communication and mutual respect between the school and the home is essential. Parents are responsible for a child's behavior. Their responsibilities include ensuring participation at Sunday mass, completion of homework assignments, and that the student attends school regularly and on-time. Parents are encouraged to be involved with the many co-curricular opportunities offered in the school and parish.
COMMUNITY-SCHOOL CONNECTION: St. Agnes is a positive presence in Little Village. It is a neighborhood school providing a faith-centered educational option for families in the area. Principles of social justice are taught through service learning projects which focus on the socioeconomic diversity of our neighborhood and world. It is important to continue to establish and utilize community-school partnerships.
CHALLENGING CURRICULUM: The field of education is constantly being reinvigorated by new research and theory, and the curriculum should evolve with those discoveries to prepare students for their future in high school and beyond. The goal of the curriculum is to create passionate and able life-long learners. The curriculum should be based on standards and emphasize a multicultural perspective. Technology should be used frequently, but with a clear purpose.
CATHOLIC IDENTITY: The school's Catholic identity is expressed through school-wide participation, especially in the traditions of the Latin American Catholic Church. Education is faith-based and gospel-centered. St. Agnes School has a strong connection with the parish maintained by the religious instruction provided by the priests and sisters.
HIGH EXPECTATIONS: Enrollment at St. Agnes School is a choice and a commitment. Graduates are expected to gain admittance to a well-reputed high school and college. Individual student needs will be met and help given, but students must be motivated and responsible for their education. Teachers and staff will hold themselves to the same high expectations that are placed on the students.
Enter event description
Reserve your pass for four days in the parish office, please bring exact amount if paying by cash.
Reserve su pase de cuatro dias en la oficina parroquial, por favor traer el dinero exacto para pagos en efectivo.
Debit/credit accepted
Last day to puchase/Ultimo dia para comprarlo Monday/Lunes, August 7th until/hasta las 7pm
If you need information please call/ nececita mas informacion llame a 773-522-0142
A community church celebration and Fundraiser includes food and beverage booths, dancing, live music stage, raffle, carnival, games and more!


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We are still dealing with issues with her weight. We watch every calorie that we can and we are pretty strict about what she eats. I hate that my 10 year old daughter has to count calories. We also try to exercise every day. She has started doing Zumba classes and she really likes them. So now we are considering this Ally's "sport" and making it a priority. Just like any other child would go to their sport practice, we are trying our best to do this with her. (well we is actually me since Jerad really wouldn't want to do this, hah!) She even got a few of her friends to go with her which makes it all that more fun.
Unfortunately, all of these attempts to mitigate her weight gain have not seemed to help much. So Dr B recommended we go to the lipid clinic at the hospital. They can do some testing of her metabolism and hormones and see if all the chemotherapy (and more likely her radiation) has affected her. I remember when doing the radiation at UC the docs there mentioned it could affect some of the glands (thyroid, pituitary, etc). Maybe there is some outlying reason why she keeps gaining weight. They will test all of these things at the clinic and hopefully be able to help us. They shared with me that 50% of ALL survivors have a weight problem throughout their life. But if that is all I have to worry about I will take it! They just want to get a little more involved so as to avoid other issues (diabetes etc). So one more area of the hospital is going to have a Barnett visit.
We were pleased to finally get the opportunity to go the Emily's Beads of Courage Dayton Dragons game. This is a game just for the Hemoc kids with their families and their nurses!! We have missed this the last few years for various reasons. I got the chance to thank Emily's mother for providing these beads for our children. Ally has "worked" really hard to earn her beads. She is immensely proud of them and so am I. I truly value them as one of our most precious things. (if the house were on fire, I would grab them!) When she wanted to take them to school to show her classmates I became very nervous. Scared they could get lost, stolen, etc. But they came home safe. Here is a picture from the game of my three heros in life wearing their beads:
Kayleigh, Caulin, and Ally.....all three ALL leukemia kids from Beavercreek!
I loved watching these kids interact at the game. They sat and compared beads and chatted. They truly have a kinship like no other. Only they can understand what it is like to go through cancer treatments as a child. They are all only about a years difference in age too. And I believe all three will grow up to be extremely compassionate people in the world and will go on to do great things. While their siblings were complaining of the cold night....no complaints from these three!
And last but not least, I wanted to let you know about a couple fundraisers. The first one is going to be at the Beavercreek Golf Club this Friday night (20th). It is a shopping event with all proceeds going towards Curesearch for these three kids. Ally's Army, Caulin's Crew, and Kayleigh's Crusaders. Please consider attending if possible. All three kids plan to be there! (I will be at the retreat for Dayton Children's but Ally should be there!) We are grateful for any money raised for this awesome organization.
Another event is being held at Charming Charlie's at the Greene Sunday night. This is for a great doctor at Dayton Children's who is running for LLS woman of the year- Dr Patricia Abboud. All the money raised goes to Leukemia and Lymphoma Society. Man/Woman of the year is the event that Ally was the honored hero for last year (and which Dad ran for the year before that). We are going to this event too.
Charming Charlie's (next to Von Maur) $10 admission, gets you 20% off your entire purchase! There will be a raffle and refreshments too! The store will open JUST FOR THE EVENT from 6pm-8pm.
Thank you Ally's Army for continuing to pray for our girl.


text: Seventeen Tamaqua High School students in grades 9 through 12 will be participating in this year's Schuylkill County Chorus which will hold at a 2 p.m. concert on Saturday, March 6, 2010 at Tri-Valley High School in Hegins. They are, from left, front row; Julian Gerace, Dana Rottet, Eileen Flamini, Erin Raymond, Kim Walker, and Drew Bonner. Second row; Laura Walsek, Emily Zancofsky, Heidi Wagner, Lauren Christ, Lauren Ward, and Marissa Guardascione. Third row; Ryan Holley, Steven Christ, Mike Kabana, Dan Clemson, Brandon Clemson, and Choral Director John Buglio.


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Zalando Gutschein
Shakespeare 400
I Capuleti e i Montecchi
L'isola disabitata
Creators & Performers
general director
board of directors
a brief history
VOICEBOX: Opera in Concert
explores the singing voice in its
purest form — stripped of sets,
costumes and accoutrements.
Our company offers a new way
of experiencing opera, one that
unleashes the expressive power of
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We showcase a repertoire of rarely
seen operatic works that resonate
with present day audiences and
provide a platform for emerging
Canadian talent. Our performances
are for music lovers who want to
expand their horizons.


text:
Jan Urbański wrote: > Heikki Linnakangas wrote: >> Jan Urbański wrote: >>> So right now the idea is to: >>> (1) pre-sort STATISTIC_KIND_MCELEM values >>> (2) build an array of pointers to detoasted values in tssel() >>> (3) use binary search when looking for MCELEMs during tsquery analysis >> >> Sounds like a plan. In (2), it's even better to detoast the values >> lazily. For a typical one-word tsquery, the binary search will only >> look at a small portion of the elements. > > Here's another version. Context diff this time, always forget to convert them... -- Jan Urbanski GPG key ID: E583D7D2 ouden estin
Next: From: Andrew Satori Date: 2008-08-14 20:33:59 Subject: API for Managing pg_hba and postgresql.conf Previous: From: Jan Urbański Date: 2008-08-14 19:45:18 Subject: Re: gsoc, oprrest function for text search take 2


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Hispanic vote: government and immigration
Every year there seems to be another push for comprehensive "immigration" reform. This is always an unpopular political venue. It's always he said she said. Let's try this again. Immigration reform. But this is especially popular when it comes time for the voting pool. Presidential candidates make a pledge. A pledge for hmm to at least talk to the League of United Latin American Citizens (LULAC). I'm just pointing out my thoughts. The LULAC has a voice.
After all the location :
National Office
2000 L Street NW
Suite 610
Washington DC 20036
Tel: (202) 833-6130 Fax: (202) 833-6135
It's right down the street from the White House. So it's not like this is a new subject with congress, senate, or the President and his cabinet. Every year all the time. There seems to be some talk, but maybe we need to ad some ear marks to the next proposition.
Then we have the Arizona State's now infamous Senate Bill 1070, which requires the police to verify the status of someone they have stopped or arrested if they suspect that the person in the country illegally. The only reason this law was enacted & passed was to mirror the Federal Law which has not been enforced very well.
The bottom line is immigration started this country, but where do you draw the line.
I understand that families and or individuals are wanting a better life. Better living conditions, food, schools, etc…. Wait with all that comes law enforcement. Some thing that was created to protect us so we could enjoy the better life; for ourselves and our family. If your here legally then there is nothing to worry about, but if your not. Not so good.
And what the couples that are here illegally and they have Anchor Babies: Part of the Immigration-Related American Lexicon. I understand the theme of a better life, but I do not agree with putting another person's life in jeopardy.
Here are some stats pulled from Poder Hispanic Magazine August/September 2010
Hispanic in the numbers:
46% of Hispanics were born in the United States.
About three-quarters of the nation's illegal immigrants are Hispanic.
32% of Hispanics were born in Mexico.
Source: AP-Univsion Polls, Pew Hispanic Center.
Israel policy may lead to deportation of kids: The country is preparing to deport children of migrant workers in an attempt to stem illegal immigration.
By Edmund Sanders
Los Angeles Times
Here is something to think about; in certain country's if your an illegal immigrant and your caught your just locked up.
Maybe one day the immigration ruling could be resolved in a manner acceptable to everyone.
Mr. President Barack Obama: Jobs, Work, and Careers
"We will go through our Federal Budget – Page by Page, line by line – eliminating those programs we don't need."
– – – President Elect Barack Obama, November 2008.
Well Mr. President it's half time and the third quarter is about to start. You are the CIC: The Commander in Chief and I don't think there has been any positive progress. Which is to bad.
Have a great day!!!
Remove yourself from the list and not receive the
or the
paper directories.
That is what this site is directed at. This is presented only in an unbiased presentation.
I have no connections to this site only to pass on information.
Muslim Electrolux workers in St. Cloud
Here is the article: 165 Muslim Electrolux workers in St. Cloud get OK to break fast for Ramadan. The workers, backed by the Minnesota chapter of the Council on American-Islamic Relations (CAIR), had charged Electrolux with violating the 1964 Civil Rights Act.
This looks more like "cry wolf." So if I go to Church every Sunday and then all of a sudden the needs of the business "XYZ Corporation" where I work decides I have to work now every Sunday. Now I can "Cry Wolf" because this is a violation of my Civil Rights. Specifically 1964 Civil Rights Act. Hmm an accommodation for me to go to Church on Sunday where I have already been going every Sunday.
When anyone applies for a job right on the application it is stated:
Are you able to perform the essential functions of the job for which you are applying, either with / without reasonable accommodation? [ ] Y or [ ] N
If no, describe the functions that cannot be performed
(Note: Company complies with the ADA and consider reasonable accommodation measures that may be necessary for eligible applicants/employees to perform essential functions. It is possible that a hire may be tested on skill/agility and may be subject to a medical examination conducted by a medical professional.)
So should every Muslim stated when Ramadan time shows up that Electrolux needs to give us reasonable accommodations? I don't think any of the Muslims would have been hired.
Maybe the company needs to build a Mosque in the complex and remove the break room. I think it would only be fair for Electrolux to give the Christians, the non-Christians, non Believers, Smokers and non-Smokers should all get reasonable accommodations. Otherwise they should also file a lawsuit againts Electrolux.
Mosque in New York City?
I love this comment from Jason Horowitz the Star Tribune – Saturday, August 21, 2010 which was actually printed in the Washington Post: By JASON HOROWITZ, Washington Post, Last update: August 20, 2010 – 9:26 PM. I will ad just one more quote from the article:– New Yorkers on both sides of the Islamic center argument are angry at outsiders who don't understand the city and fan the flames: "They are trying to create propaganda." The Article comes from the
Newt Gingrich has argued, among other things, that the Muslim congregation shouldn't build the center because "Nazis don't have the right to put up a sign next to the Holocaust Museum in Washington." Sarah Palin has weighed in, too, in opposing the "ground zero mosque." The pain she said, is "too raw, too real."
Now I would like to know why after all these years since September 11, 2001 that, now a religious group who is associated with Terrorism in the United States is pushing to put up a Mosque in the vicinity of the Terrorist Attack that took place. What and why is this location so important? Why now?
Who wants to be the newbie in charge?
"FOR THE PEOPLE ABOUT THE PEOPLE" Another four years is about to come to an end; for our current President. "But what has he and his cabinet done for Our Country?" So the arena has opened up to new candidates. A new aura into the future of our country. Every candidate will bring something to the table, but I ask who wants more than one helping?
Nothing has been done in Government except for each political party just points the finger at each other. Who did this and who did that. Corruption at it's finest. But wait it is all on the Government level. Lawyers make rules to protect themselves and no one else.
Wow!! Have a nice day.


text:
Tweets from @TBHomeTeam/writers
MORE from our HomeTeam writers.
Tue. November 20, 2012 | Times Staff
Ryan Orr, the Tampa Bay Times' Golfer of the Year last season, signed recently with the University of Florida.
"I'm very excited to have the opportunity to become a Gator and play for Buddy Alexander, one of the finest college golf coaches in the country," said Orr, who shot a two-day total of 156 at the Class A state tournament this season to finish tied for 13th, the highest finisher among bay area boys.
The senior, also the region champion, won the Florida Junior Tour Innisbrook tournament in May, qualified for the U.S. Junior Amateur and tied for third at the Boys Junior Championship at TPC Sawgrass.


text:
- 1 teaspoon unflavored gelatin
- 1/2 cup sugar
- 1/4 cup fresh lime juice
- 1 teaspoon finely grated lime zest
- 2 pints strawberries, hulled and thinly sliced
- 1/2 vanilla bean, split lengthwise and seeds scraped
- 1/2 cup heavy cream, chilled
- Vanilla ice cream, for serving
Set 4 martini glasses or shallow dessert bowls in the freezer. In a small cup, sprinkle the gelatin over 1 tablespoon of cold water and let stand until softened, about 3 minutes. Fill a medium bowl half full of ice water.
In a small saucepan, combine 1/4 cup of the sugar with 2 tablespoons of water and simmer over moderate heat just until the sugar is dissolved. Remove the sugar syrup from the heat and whisk in the softened gelatin until dissolved. Stir in the lime juice and 1/2 teaspoon of the lime zest. Set the saucepan in the ice water bath and stir the gelée until cooled. Pour the gelée into the chilled glasses and freeze until firm, about 15 minutes.
Meanwhile, in a medium saucepan, combine 1 cup of the strawberries with 2 tablespoons of sugar and 1/4 cup of water and bring to a boil. Let cool slightly. Transfer the strawberry mixture to a blender and puree until smooth. Pour the puree into a bowl and stir in the remaining strawberries and half of the vanilla bean seeds; let cool.
In a medium bowl, whisk the heavy cream with the remaining 2 tablespoons of sugar, 1/2 teaspoon of lime zest and vanilla bean seeds until soft peaks form. Spoon the strawberry puree over the lime gelée. Top the strawberry puree with a scoop of vanilla ice cream and a dollop of the whipped cream and serve immediately.


text:
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text:
|Date added:
|May 17, 2012
|Downloads last week:
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text:
Maxi Taxi Melbourne
Affordable Chauffeur Option - Get Quote Now
Your browser does not support iframes.
Maxi Taxi Melbourne Companies
Although they may refer to them in terms other than Maxi Taxi Melbourne, the following taxi service companies each offer vehicles which can carry a
greater amount of passengers which we refer to as Maxi Taxi Melbourne
Dandenong Taxis - Phone on 9791 2111
Frankston Taxis - Phone on 9786 3322
13 CABS - Phone on 13 22 27
Silver Top Taxis - Phone on 13 10 08
Why a Maxi Taxi in Melbourne
When wanting to book any sort of ground transportation in Melbourne, the choice of whether to book Maxi Taxi or a regular Taxi Cab is quite simple. The Maxi Taxi, although can be more costly, does allow more passengers to travel in the vehicle.
A Maxi Taxi in Melbourne can also allow for the passengers to carry more luggage in the vehicle, but off course this is dependent on the size of the Maxi Taxi.
Another option to consider is whether to book a Maxi Taxi Melbourne or a chauffeured option of a limousine transfer. This is generally a difficult decision to make because both of them offer great options. We will discuss these two options further below.
Maxi Taxi or Chauffeured Limo?
When wanting to travel with more than four passengers in and around Melbourne, the options are multiple. There is always the option of public transport with Trams or Trains or Buses, however for a more personal and private experience it is often a decision between a Maxi Taxi or a Chauffeured Van/People Carrier.
A Maxi Taxi could be cheaper to travel than in a chauffeured option which is a great advantage. In terms of pricing, the only downfall of Maxi Taxi Melbourne services is that the prices are not fixed and are determined by a Meter inside the vehicle. The price is dependent upon the Kilometers which are driven, the time taken in the transfer, road tolls encountered, the amount of passengers and the flag fall fee of the vehicle. All of these factors are used to make up the price of a single Maxi Taxi transfer in Melbourne.
Despite all this, in most cases it will still be cheaper to travel in a Maxi Taxi than a chauffeured option. Chauffeured People Carrier companies, such as Driven By Limo, do offer fixed price quote estimates for their transfers however the prices are excluding road tolls. The advantages of luxury chauffeured van transfers in Melbourne are generally that the chauffeur is more professional, presentable and dedicated to your luxury service. The chauffeured vehicles are most commonly also more modern vehicles and well maintained to be kept in a crisp condition.
24 Hours Approval Directory
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Privacy Statement
Terms Of Use
Copyright (c) 2012
Maxi Taxi Melbourne


text:
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Search by
Back to School Jam
This event has already taken place. Click here for the latest events.30-08-2013 12:00:00 30-08-2013 12:00:00 America/New_York Back to School Jam | 11am-2pm Celebrate going back-to-school or starting school for the very first time with an afternoon of fun activities. Children will enjoy a fun-filled event with crafts, snacks, music, giveaways, and more. This celebration helps young children look forward to a new school year and another important stage of learning. http://www.nymetroparents.com/2015neweventinfo.cfm?id=141135 Brooklyn Children's Museum Brooklyn Children's Museum
If you go: Please tell them you saw this event on
Venue Description:
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text:
Garbage and Recycling Collectors
- Think about what you trash. Cat litter improperly disposed of is a pet peeve, says Terrance Flynn, a New York City sanitation worker. "People put it in a bag hidden inside another bag," so unsuspecting workers don't see it, says Flynn. The blade that scoops up trash often breaks the bags, spraying the litter everywhere.
- Don't overload containers. Americans create an average of 4.6 pounds of garbage per person a day, and some routes cover up to 1,000 homes. Heavy cans and bins can cause muscle sprains.
- Never mix recyclables and trash. This can be illegal, plus "it's a nuisance, and it costs the city money," says Flynn.
- Watch where you put the bins. Don't leave them where dogs might think they're trees or fire hydrants (blocking hydrants is a safety hazard, too).


text:
As part of Pepsi "Refresh your world" project, we asked 20 young Czech artists to "refresh" the famous "Slavic Epopee" by Alfons Mucha. Artists interpreted the original paintings to reflect different issues facing young people today.
Print advertisment created by BBDO, Czechia for Pepsi, within the category: Non-Alcoholic Drinks.
Creative Director: Sndrej Stuk
Art Directors: Ales Kolaja, Lubos Vacke
Copywriters: Jan Holecek, Iva Macku
Digital: Martin Kermes / 2fresh
Account Director: Mirka Korbelova
Account Manager: Sarka Sudova
Production: Jan Soupa, Savage
Executive producer: Jan Daczicky
Producer: Michaela Berkova
Director: Daniel Bird
Highest Rated
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"creativity is what I think in every moment of my life"
www.facebook.com/manzoordsign I www.behance.net/manzoordesign
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Pretty much what we expected: Carnival Freedom Cruise Review by tubaman77
Overall Member Rating
Pretty much what we expected
Destination: Western Caribbean
Embarkation: Cozumel
The buffet foods were adequate, with the best food being from the burger joint. Somehow though going on a cruise and finding the best food to be a burger is somewhat of a let down. On the other hand, the dining room service and food was quite good, yet nothing near the quality of some of the competition out on the seas. There is some evidence of the Holland America influence in some of the dishes and flavors so More they do have that going for them. Cabins, as Carnival has always had, are some of the most spacious available, and are certainly comfortable and clean. The bathrooms are certainly larger than most on many other ships and I actually didn't have to go outside to change my mind. I will say though that the bedding was some of the most uncomfortable I've ever experience on a ship. I guess they figure though that most people on a Carnival cruise won't sleep much anyways, so what's the difference. Which brings me to my biggest and most annoying complaint.
Carnival markets themselves as having non-smoking facilities on their ships, but unless one reads all the fine print up front, which you'd think I would have done having traveled as much as I have, one won't realize that yes, most of the public areas are non-smoking, but smoking is allowed in some of the most annoying and frequently trans versed areas of the ship. That in itself one can work around as long as you don't care about going into the casino or any of the lounges surrounding it, or on the port side of the Lido Deck and the bar in that area, but what we learned much to our chagrin was that smoking is still allowed on the balconies of the cabins that have balconies, which on this ship make up the vast majority of the cabins. We had the pleasure of having the most obnoxious people next to us who were not only loud and inconsiderate throughout the entire cruse, but must have brought some sort of cow feces soaked rope along to smoke that would make most any of God's creatures gag - including those smoking it. If we were on the balcony and they came outside we knew we had less than 10 seconds to get inside and close the door tightly before we would be gassed. Then we would get to listen to them hack, cough, choke, puke, spit, and we think, vomit while smoking that vile material. The bottom line is, don't bother paying for a balcony if you are a non-smoker on Carnival's ships. You're wasting your money because you won't be able to enjoy it without having the ocean smell like an ash tray.
So all that being said, the cruise product itself is fine, and even with just under 3000 passengers on board, you'd really never know there were that many people surrounding you. But unfortunately the line markets itself as a mass market product, and wants those smokers onboard as they are the ones who will spend the money in the casinos and bars. No wonder I've always like RCCL and Princess better... Less
Published 12/29/13
Cabin review: 8583
Clean, comfortable, spacious, pleasantly lighted, spacious balcony. Bedding quite uncomfortable.
Port and Shore Excursions
As we have visited Cozumel several times over the years, we have been able to distance ourselves from the crowds and find parts of the island that we can have all to ourselves. We 've rented Jeep wranglers the last couple times there and driven around to the east side of the island to view the spectacular scenery and to be able to sit on a 5 mile long beach all by ourselves. The water is spectacular in color, there are blow holes galore, and the sand is like powder. And we've found a little family run shop in the jungle that sells some of the best jewelry on the island along with lots of other touristy stuff at prices far below what they are by the ships. Take the chance - get away from the crowds. Just don't try to drive in town....
Tendering to the island from the ship in a driving rain storm is quite and experience, but it just ads to the adventure. Once we were on land we just waiting out the rain under one of the tents and then walked in town to check out some of the shopping. It was OK. Much of the same stuff one can find in the US if you look hard enough, or even sit in the living room and buy it from QVC. And prices really aren't all that great either. Bermuda is still the shopping value center.
Once we were tired of shopping and the weather dried up some more, we simply jumped on one of the island busses and road up 7 Mile Beach to the Governors Residence and plopped ourselves in the sand for a great day of quiet beach time again far from the crowds. Snorkeling right off the beach there turned out to be better than what those who paid many dollars to do on a preplanned excursion experienced, and we did it all including rented snorkel equipment and bus fare for $20. Cayman is all about the beach. Still one of the best.
Read Cruise Critic's Carnival Freedom Review >>
Older Ship, Great Staff, Easy ...
Exceeded our expectations for ...
First Cruise Ever!
First Cruise
This was awesome!! This was ou...
Enjoyed our cruise. The CD was...
Older Ship, Great Staff, Easy ...
Exceeded our expectations for ...
First Cruise Ever!
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