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Duchenne Muscular Dystrophy (DMD) is a fatal muscle wasting disease manifested in young boys, for which there is no current cure. We have shown that the amino acid taurine is safe and effective at preventing dystropathology in the mdx mouse model for DMD. This study aimed to establish if treating growing mdx mice with a higher dose of taurine was more effective at improving strength and reducing inflammation and oxidative stress. Mice were treated with a dose of taurine estimated to be 16 g/kg/day, in drinking water from 1–6 weeks of age, after which in vivo and ex vivo muscle strength was assessed, as were measures of inflammation, oxidative stress and taurine metabolism. While the dose did decrease inflammation and protein oxidation in dystrophic muscles, there was no improvement in muscle strength (in contrast with benefits observed with the lower dose) and growth of the young mice was significantly restricted. We present novel data that a high taurine dose increases the cysteine content of both mdx liver and plasma, a possible result of down regulation of the taurine synthesis pathway in the liver (which functions to dispose of excess cysteine, which is toxic). These data caution that a high dose of taurine can have adverse effects and may be less efficacious than lower taurine doses. Therefore, monitoring of taurine dosage needs to be considered in future pre-clinical trials, in anticipation of using taurine as a clinical therapy for growing DMD boys (and other conditions).
Citation: Terrill JR, Pinniger GJ, Nair KV, Grounds MD, Arthur PG (2017) Beneficial effects of high dose taurine treatment in juvenile dystrophic mdx mice are offset by growth restriction. PLoS ONE 12(11): e0187317. https://doi.org/10.1371/journal.pone.0187317
Editor: Jozef Dulak, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, POLAND
Received: July 17, 2017; Accepted: October 17, 2017; Published: November 2, 2017
Copyright: © 2017 Terrill 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.
Data Availability: All relevant data are within the paper and its Supporting Information files.
Funding: This research was funded by the National Health and Medical Research Council of Australia (https://www.nhmrc.gov.au), grant number APP1065829 (PGA). 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.
Duchenne Muscular Dystrophy (DMD) is a lethal, X-chromosome linked muscle disease affecting about 1 in 3500–6000 boys worldwide (Reviewed in [1, 2]). DMD is caused by mutations in the dystrophin gene resulting in the loss of functional dystrophin protein in muscle, which leads to increased susceptibility of sarcolemmal damage and necrosis after muscle contraction [3, 4]. Consequently, dystrophic muscle undergoes repeated cycles of myofibre necrosis, associated with subsequent inflammation, fibrosis and regeneration: where this damage persists over many months and years in DMD boys it results in the replacement of muscle with fat and fibrotic tissue [5, 6]. The severe loss of muscle mass and function in DMD boys leads to premature death often due to respiratory or cardiac failure (reviewed in [1, 7]). While much effort has been dedicated to developing genetic or molecular therapies to replace the missing dystrophic protein [8, 9], there is no cure yet for DMD: the standard drug treatment using corticosteroids is limited in efficacy and associated with severe side effects [10, 11]. Consequently, there is considerable interest in developing safe and efficacious therapeutic drug or nutritional interventions that can prevent or slow the progression of DMD (reviewed in [12–14]).
The semi-essential amino acid taurine (2-aminoethanesulfonic acid) has been investigated as a pharmacological intervention, using the mdx mouse model of DMD [15–21]. In these studies, administration of taurine to mdx mice prevented myofibre necrosis, improved both in vivo and ex vivo muscle strength, increased fatigue resistance and decreased other indices of mdx pathology such as inflammation and oxidative stress. Recently we showed that an oral dose of approximately 4 g/kg/day (2% in drinking water, from 18 days until 6 weeks of age) had benefits on many indices of dystropathology (with improved in vivo and ex vivo muscle strength and decreased inflammation and oxidative stress) in mdx mice . Research by De Luca and colleagues in 2003 established that treating male mdx mice aged 4 weeks (until 8 weeks) with a high dose of taurine (up to 400 mg per mouse per day, equivalent to about 16 g/kg/day calculated for a mouse weighing 25 g) was highly effective at reducing exercise induced muscle weakness .
We propose that one important mechanism of action of taurine in protecting mdx muscle from necrosis relates to effects on neutrophils (inflammatory cells that rapidly arrive at sites of damage). Specifically, taurine scavenges hypochlorous acid (HOCl), a highly reactive oxidant that is produced by the interaction of hydrogen peroxide and myeloperoxidase (MPO), a heme protein secreted by neutrophils . HOCl can cause irreversible oxidative damage to proteins, as well as cause reversible protein thiol oxidation, which can dramatically modify protein function . The scavenging of HOCl by taurine results in the formation of taurine chloramine, a molecule that itself exerts anti-inflammatory effects such as inhibiting the production of pro-inflammatory cytokines and nitric oxide (NO), and the inhibition of NF-κB activation by the oxidation of IκB-α [22, 24]. We have previously shown that muscle from the Golden Retriever Muscular Dystrophy (GRMD) dog model of DMD undergoes excessive neutrophil infiltration, leading to increased MPO content and HOCl production, which is associated with increased protein oxidation . Likewise, we show that increased MPO content and protein thiol oxidation in mdx muscle is ameliorated in mice treated with a taurine dose of 4 g/kg/day (as discussed above) . Therefore, the effects of a higher dose of taurine, estimated to be about 16 g/kg/day, on neutrophil and MPO content and protein thiol oxidation in mdx muscle were examined in the current study.
The aim of this study was to test if a high dose of taurine (16 g/kg/day) in young growing mdx mice was highly effective at reducing dystropathology, specifically for improving muscle strength and decreasing inflammation and protein thiol oxidation and to discern if these benefits were associated with decreased neutrophil and MPO content. Since we have shown previously that taurine metabolism (including synthesis of taurine in the liver from cysteine) is perturbed in mdx mice , we also assessed the effects of this high taurine dose on the content of taurine and cysteine in the liver, plasma and muscle of mdx mice, and examined taurine synthesis in the liver.
Materials and methods
All reagents were obtained from Sigma-Aldrich unless otherwise specified.
All animal experiments were conducted in strict accordance with the guidelines of the National Health and Medical Research Council Code of practice for the care and use of animals for scientific purposes (2004) and the Animal Welfare act of Western Australia (2002) and were approved by the Animal Ethics committee at the University of Western Australia.
All experiments were carried out on dystrophic mdx (C57Bl/10ScSnmdx/mdx) and non-dystrophic control C57 (C57Bl/10ScSn) mice (the parental strain for mdx). Mice were obtained from the Animal Resource Centre, Murdoch, Western Australia. Mice were maintained at the University of Western Australia on a 12-h light/dark cycle, under standard conditions, with free access to food and drinking water.
Mothers (and pups) were given either no treatment or taurine (delivered by 8% in the drinking water) from 7 days after birth, and taurine treatment continued after weaning (at 21 days) until they were sampled at 6 weeks. There were 9 mice in the C57 group, 10 in the untreated mdx group and 8 in the taurine treated group, however 8 mice of each group were randomly chosen for biochemical analysis. Several mice were removed from the ex vivo force measurements because the mice died under anaesthesia before the muscle could be removed. Both male and female pups were used in the study with equal numbers of both gender in each group. There were no significant effects of gender observed, except for liver (C57) and quadriceps (untreated mdx) weights (see S1 Table). Water ingestion and body weights were monitored twice weekly for all mdx treatment groups. There was no difference in water consumption of untreated and taurine treated mdx mice (averaging 3.2±0.5 and 4±2.3 ml per mouse per day respectively). The approximate consumption of taurine, based on water consumption, was about 16 g/kg/day.
The grip strength of all mice was measured using a Chatillon Digital Force Gauge (DFE-002) and a triangle metal bar, as per the TREAT-NMD recommended standard protocol “Use of grip strength meter to assess limb strength of mdx mice– M.2.2_001 “http://www.treat-nmd.eu/downloads/file/sops/dmd/MDX/DMD_M.2.2.001.pdf”. In brief, the mouse was placed on the front of the triangle bar (attached to a force transducer) and pulled gently until released. Each mouse underwent 5 consecutive grip-strength trials; the grip strength value for each mouse was recorded as the average of the three trials with the highest force. Average grip strength was presented as total force (g) and also normalized for body weight [Force (g)/BW (g)]. Grip strength measurements were performed on three separate occasions during the final week of treatment to accustom mice, however only data from the final testing session was used for analysis.
Mice (at 6 weeks of age) were anaesthetized via an intraperitoneal (IP) injection of sodium pentobarbitone (40 mg/kg body weight). Anaesthetized mice were placed on a heated plate at 37°C to maintain core body temperature. Following dissection of the EDL muscle (for physiological measurements of contractile function), blood was removed via cardiac puncture and other tissues, including the quadriceps muscle and liver, were removed and snap frozen for further analysis.
Ex vivo muscle function testing was performed as per . In brief, the EDL tendons were tied with non-absorbable black braided surgical silk (Dysilk, Dynek Pty Ltd) and mounted in an in vitro muscle test system (805A In Vitro Force Transducer System, Aurora Scientific Inc.) filled with mammalian ringer solution (containing (in mM) NaCl (121); KCl (5.4); MgSO4.7H2O (1.2); NaHCO3 (25); HEPES (5); glucose (11.5) and CaCl2 (2.5)), bubbled with Carbogen and maintained at 25°C. The muscle was stimulated with 0.2 ms square wave pulses (701B High Power Bi-phase Current Stimulator, Aurora Scientific Inc.) via parallel electrodes situated on both sides of the suspended muscle.
Ex vivo measurement of force
At the start of each experiment the muscle was set to the optimal length (Lo) by manually adjusting the lever arm and recording the twitch force response. The length at which the maximum twitch force was recorded was taken as Lo. All subsequent measurements of contractile function were performed at Lo. The optimal fibre length (Lf) in the EDL was calculated from a pre-defined fibre-length to muscle-length ratio of 0.44 . The force frequency relationship was evaluated by recording the force responses at stimulation frequencies of 5, 10, 20, 30, 50, 80, 100, 120 and 150 Hz. Each stimulus was separated by a two minute interval to avoid inducing muscle fatigue. The stimulation that produced the greatest force throughout the experiment was recorded as the maximum tetanic force.
Force recordings were normalized relative to muscle cross sectional area (CSA) and presented as specific force (N/cm2). CSA was estimated by dividing the wet muscle mass (g) by Lf and the density of mammalian skeletal muscle (1.056 mg / mm3).
Neutrophil presence in quadriceps muscles was measured using immunoblotting, as described below. MPO catalyses the production of hypochlorous acid from hydrogen peroxide and chloride  and hypochlorous acid reacts with 2-[6-(4-aminophenoxy)-3-oxo-3H-xanthen-9-yl]benzoic acid (APF) to form the highly fluorescent compound fluorescein, that is measured in this method, as described previously . Briefly, frozen muscle was ground using a mortar and pestle under liquid nitrogen and homogenised in 0.5% hexadecyltrimethylammonium bromide in PBS. Samples were centrifuged and supernatants diluted in PBS. Human MPO was used as the standard for the assay (Cayman Chemical). Aliquots of each experimental sample or MPO standard were pipetted into a 384 well plate, before the addition of APF working solution (20 μM APF and 20 μM hydrogen peroxide in PBS) was added. The plate was incubated at room temperature (protected from light) for 30 minutes, with fluorescence measured every minute using excitation at 485 nm and emission at 515–530 nm. The rate of change of fluorescence for each sample was compared to that of the standards and results were expressed per mg of protein, quantified using the DC protein assay (Bio-Rad).
Quantification of protein thiol oxidation
Reduced and oxidized protein thiols were measured in quadriceps muscle using the 2 tag technique as described previously . In brief, frozen tissue was crushed under liquid nitrogen, before protein was extracted with 20% trichloroacetic acid (TCA)/acetone. Protein was solubilized in 0.5% sodium dodecyl sulphate (SDS) with 0.5 M Tris at pH 7.3 (SDS buffer) and protein thiols were labeled with the fluorescent dye BODIPY FL-N-(2-aminoethyl) maleimide (FLM, Invitrogen). Following removal of the unbound dye using 100mM cysteine, oxidized thiols were reduced with tris(2-carboxyethyl)phosphine (TCEP) before the subsequent unlabeled reduced thiols were labeled with a second fluorescent dye Texas Red C2-maleimide (Texas red, Invitrogen). The sample was washed in acetone and re-suspended in SDS buffer. Samples were read using a fluorescent plate reader (Fluostar Optima) with wavelengths set at excitation 485 nm, emission 520 nm for FLM and excitation 595 nm, emission 610 nm for Texas red. A standard curve for each dye was generated using ovalbumin and results were expressed per mg of protein, quantified using the DC protein assay (Bio-Rad).
HPLC analysis of taurine and cysteine
Taurine and cysteine levels in liver, plasma and quadriceps muscle were measured using reverse phase high performance liquid chromatography (HPLC) as previously described . In brief, frozen tissues were crushed using a mortar and pestle under liquid nitrogen and homogenized in 25 times 5% TCA. Plasma samples were precipitated by addition of 10 times by volume of 5% trichloroacetic acid (TCA). After centrifugation, supernatants were removed and stored at -80°C until analysis. Analytes were separated using HPLC with fluorescent detection, with pre-column derivatisation with o-phthalaldehyde (OPA) and 2-mercaptoethanol (2ME). OPA reacts rapidly with amino acids and sulfhydryl groups to yield intensely fluorescent derivatives, and 2ME, a reducing agent, prevents the OPA reagent from oxidising. Supernatants were mixed with iodoacetamide, dissolved in 5% TCA, to a final concentration of 25 mM. An internal standard, o-phospho-dl-serine, dissolved in 5% TCA was added to a final concentration of 5 mM. Sodium borate was used to adjust the pH to 9. Samples were placed in an autosampler, which was maintained at 4°C. Samples were mixed on a sample loop with a derivitising solution containing 40 mM OPA and 160 mM 2ME in 100 mM sodium borate, pH 12, for 30 seconds before injection onto the column. Separation was achieved with a C18 column (5 μl, 4.6 x 150 mm, Phenomenex) using a Dionex Ultimate 3000 HPLC system. Mobile phase A consisted of 50 mM potassium phosphate buffer and tetrahydrofuran (97:3). Mobile phase B consisted of 90% methanol, with a gradient increase in B from 0 to 25%. Fluorescence was set at 360 nm and 455 nm for excitation and emission respectively. The protein content of liver and muscle samples were quantified by solubilising the pellet in 0.5 M sodium hydroxide, before incubation at 80°C for 15 minutes. Once fully dissolved, protein concentrations of supernatants were quantified using a Bradford protein assay (Bio-Rad).
Protein extraction and immunoblotting
Frozen livers and quadriceps muscles were crushed using a mortar and pestle under liquid nitrogen and homogenized in 1 ml ice-cold 1% NP40, 1 mM EDTA in phosphate buffered saline (PBS), supplemented with complete EDTA free protease inhibitor tablets and PhosSTOP phosphatase inhibitor tablets (Roche), and centrifuged at 10,000 g for 10 min. The protein concentrations of supernatants were quantitated using the DC protein assay (Bio-Rad). Samples were resolved on 4–15% SDS-PAGE TGX Stain-Free gels (Bio-Rad) and transferred onto nitrocellulose membrane using a Trans Turbo Blot system (Bio-Rad), after imaging for total protein using the Stain-Free imaging program on the ChemiDoc MP Imaging System (Bio-Rad). Immuno-blotting was performed with rabbit polyclonal antibodies to neutrophil elastase (ab68672, Abcam), cysteine dioxygenase type 1 (ab53436, Abcam) and cysteine sulfinate decarboxylase (ab101847, Abcam) all dissolved 1:1000 in 5% bovine serum albumin (BSA). HRP-conjugated goat anti-rabbit secondary antibodies were from Thermo Fisher Scientific. Chemiluminescence signal was captured using the ChemiDoc MP Imaging System (Bio-Rad). Resultant images were quantified using ImageJ software . A common sample was loaded onto each gel to normalise for detection efficiencies across membranes, and all bands were standardised to total protein for that lane .
Cysteine dioxygenase activity
Cysteine dioxygenase activity was measured by quantification of cysteine sulfinate from substrate cysteine, as per , with some modifications. In brief, frozen livers were crushed using a mortar and pestle under liquid nitrogen and homogenized in 10 times 200 mM 2-(N-Morpholino)ethanesulfonic acid (MES) at pH 6.1. After centrifugation, supernatants were removed and aliquoted into two tubes (one labelled blank). Remaining supernatant was retained and the protein concentration quantitated using the DC protein assay (Bio-Rad). To both other tubes, a mixture containing 2.5 mM ferrous sulphate, 10 mM NAD+ and 25 mM hydroxylamine hydrochloride at pH 5.9 was added. To both tubes, a mixture containing 40 mM cysteine and 0.50 mM bathocuproinedisulfonic acid was added. 20% sulfosalicylic acid was immediately added to the blank tubes, and all samples were incubated for 5 minutes. 20% sulfosalicylic acid was added to the rest of the tubes, and samples were incubated on ice for 15 minutes before centrifugation to remove the precipitated protein. Supernatants were removed and stored at -80°C before analysis. Analytes were separated using HPLC with fluorescent detection, with pre-column derivitisation with OPA and 2ME as per analysis of taurine and cysteine as above. Activity of cysteine dioxygenase was determined to be the amount of cysteine sulfinate formed per minute, by subtracting the cysteine sulfinate content of the sample with cysteine substrate added minus the cysteine sulfinate content of the sample without cysteine substrate, divided by 5 (samples were incubated for 5 minutes).
Data were analysed using GraphPad Prism software. One-way ANOVA tests with post-hoc (LMS) comparisons were used to identify significant differences between experimental groups. Two-way repeated measures ANOVAs were used to analyse data for the force-frequency protocols. Statistical significance was accepted at p<0.05. Phenotype and force data are presented as mean ± SEM, all data points are presented ± SEM for all other measures.
Body, muscle and liver weights were measured at 6 weeks of age to identify potential detrimental effects of treatment with taurine in young growing mdx mice. While body weights were similar for C57 and untreated mdx mice, taurine treatment reduced mdx body weight by 22% (Table 1). Liver weights were similar across all groups. Quadriceps and EDL muscles were heavier in untreated mdx mice compared with C57 mice (44% and 18% respectively). Taurine treatment reduced mdx quadriceps and EDL muscle weight by ~30% (Table 1). Tibia length was similar for C57 and mdx mice (Table 1), and taurine treatment reduced mdx tibia length by 12%. EDL cross sectional area (to which force recordings were normalised) was similar for C57 and mdx mice (Table 1), however, taurine treatment reduced the mdx EDL cross sectional area by 25%. These data indicate that treating juvenile mdx mice with a high dose of taurine (16 g/kg/day) causes growth restriction.
Grip strength and ex vivo muscle strength
The grip strength test is a non-invasive in vivo measure of mouse limb strength. Mdx mice had weaker (15%) normalised (to body weight) grip strength (Fig 1A) and total grip strength (Fig 1B), compared with C57 mice. Although taurine treatment of mdx mice had no effect on normalised grip strength (Fig 1A), there was a 15% reduction in total grip strength (Fig 1B).
Data are presented as both force normalised to body weight (A) and total force (B). Symbols for significant differences (p<0.05) are- * = between untreated mdx and C57, $ = between untreated mdx and taurine treated mdx and # between taurine treated mdx and C57. Data are presented as mean ± SEM and n = C57 (9), mdx (10) and taurine treated (8).
Mdx EDL muscle produced lower specific tetanic forces ex vivo at stimulation frequencies of 30 Hz and above (Fig 2A), and lower total tetanic forces at stimulation frequencies of 40 Hz and above (Fig 2B), compared with C57 EDL muscle. While taurine treatment had no effect on specific tetanic force of mdx EDLs (Fig 2A), there was a reduction in total tetanic force at stimulation frequencies of 40 Hz and above (Fig 2B). These data show that the high taurine dose did not improve either in vivo or ex vivo muscle strength.
Isometric force produced at frequencies from 5–150 Hz was plotted for both specific force (A) and total force (B). Symbols for significant differences (p<0.05) are- * = between untreated mdx and C57, $ = between untreated mdx and taurine treated mdx and # between taurine treated mdx and C57. Data are presented as mean ± SEM and n = C57 (7), mdx (9) and taurine treated (7).
Inflammation and protein thiol oxidation
Neutrophil content was 250% higher in mdx muscle compared with C57 muscle (Fig 3A), and taurine treatment reduced, by 70%, neutrophil content of mdx muscle back to C57 levels. MPO content was 440% higher in mdx muscle compared with C57 muscle (Fig 3B), and taurine treatment reduced, by 50%, MPO content of mdx muscle. Protein thiol oxidation was 20% higher in mdx muscle compared with C57 muscle (Fig 3C), and taurine treatment reduced, by 20%, protein thiol oxidation in mdx muscle, back to levels in C57 mice. Overall, the high dose taurine had striking benefits on these parameters in the dystrophic mdx muscles.
Quadriceps muscle content of neutrophil elastase (A) and MPO (B) and percentage of protein thiol oxidation (C) in untreated C57, untreated mdx and taurine treated mdx mice. Symbols for significant differences (p<0.05) are- * = between untreated mdx and C57, $ = between untreated mdx and taurine treated mdx. Data are presented as mean ± SEM and n = C57 (8), mdx (8) and taurine treated (8). A representative blot for neutrophil elastase is shown, protein was standardised to total protein using stain-free gels. From the same stain-free gel image, the loading of actin is shown.
Taurine and cysteine content of liver, plasma and muscle
Taurine content was similar for C57 and mdx liver (Fig 4A), and taurine treatment led to a 200% increase in taurine content of mdx liver. Taurine content was similar for C57 and mdx plasma (Fig 4C), and taurine treatment increased taurine content of mdx plasma by 1100%. Muscle taurine content was similar for all groups (Fig 4E). Cysteine content was 20% lower in mdx liver compared with C57 (Fig 4B), and taurine treatment increased, by 30%, cysteine content of mdx liver, back to C57 levels. Cysteine content was 35% lower in mdx plasma compared with C57 (Fig 4D), and taurine treatment increased cysteine content of mdx liver by 100%, with cysteine content of taurine treated mdx plasma being also 33% higher than C57 plasma. Muscle cysteine content was similar for C57 and mdx groups; however cysteine content of taurine treated mdx muscle was 30% lower that C57 muscle (Fig 4F).
Liver content of taurine (A) and cysteine (B), plasma content of taurine (C) and cysteine (D) and quadriceps muscle content of taurine (D) and cysteine (E) in untreated C57, untreated mdx and taurine treated mdx mice. Symbols for significant differences (p<0.05) are- * = between untreated mdx and C57, $ = between untreated mdx and taurine treated mdx and # between taurine treated mdx and C57. Data are presented as mean ± SEM and n = C57 (8), mdx (8) and taurine treated (8).
These data indicate that delivery of the high dose of taurine (16 g/kg/day) in the drinking water to mdx mice is highly effective at increasing levels of taurine in the liver and plasma; however, this high dose taurine causes a (potentially toxic) build-up of cysteine.
Liver cysteine dioxygenase and cysteine sulfinate decarboxylase
Immunoblotting quantification of protein levels of enzymes related to cysteine metabolism showed that cysteine dioxygenase protein was 140% higher in livers of untreated mdx mice compared with C57 (Fig 5A), and taurine treatment reduced, by 60%, cysteine dioxygenase content of mdx liver back to levels of C57 liver. Measurement of enzymatic activity showed that cysteine dioxygenase activity was 60% higher in mdx liver compared with C57 (Fig 5B), and taurine treatment dramatically decreased, by 80%, the cysteine dioxygenase activity of mdx liver, to well below the activity of C57 livers. Cysteine sulfinate (the metabolite intermediary between cysteine and taurine) content was similar for all groups (Fig 5C). Cysteine sulfinate decarboxylase was 60% lower in mdx liver compared to C57 liver (Fig 5D), and taurine treatment reduced cysteine sulfinate decarboxylase content of mdx liver by 97%.
Liver content (A) and activity (B) of cysteine dioxygenase, content of cysteine sulfinate (C) and cysteine sulfinate decarboxylase (D) in untreated C57, untreated mdx and taurine treated mdx mice. CD = cysteine dioxygeasne, CS = cysteine sulfinate, CSD = cysteine sulfinate decarboxylase. Symbols for significant differences (p<0.05) are- * = between untreated mdx and C57, $ = between untreated mdx and taurine treated mdx and # between taurine treated mdx and C57. Data are presented as mean ± SEM and n = C57 (8), mdx (8) and taurine treated (8). Representative blots are shown, proteins were standardised to total protein using stain-free gels. From the same stain-free gel image, the loading of albumin is shown.
These data show that treating juvenile mdx mice with a high dose of taurine likely causes a down-regulation of taurine synthesis in mdx liver, leading to a build-up of cysteine in mdx liver and plasma.
The key novel observation in this study is that delivery of high dose taurine in drinking water, estimated to be about 16 g/kg/day, caused growth deficits in young mdx mice. We propose that the cause of the growth deficit is an indirect consequence of an increase in cysteine levels resulting from an inhibition of the taurine synthesis pathway (which disposes of excess cysteine) in mdx mice. Nevertheless, consistent with previous work, there was good evidence that taurine decreased inflammation and protein oxidation in dystrophic tissue.
We and others have shown that taurine treatment of mdx mice (with doses ranging from 1 to 16 g/kg/day) decreases dystropathology, with an improvement in muscle strength and a decrease in myofibre necrosis, inflammation and oxidative stress [15–21]. Likewise, we show in the current study using young juvenile mdx mice that a high taurine dose of 16 g/kg/day (from 7 days until 6 weeks of age) decreased neutrophil and MPO content of mdx muscle, as well as protein thiol oxidation, supporting our hypothesis that taurine exerts strong anti-inflammatory and antioxidants benefits in dystrophic muscle. We propose a likely mechanism involves scavenging of HOCl by taurine, and subsequent formation of the anti-inflammatory molecule taurine chloramine [22, 24, 25].
The high dose taurine in the present study resulted in a decrease in total in vivo and ex vivo force. However this dose did not affect in vivo and ex vivo muscle strength when normalised to body weight (grip strength) or to cross sectional area calculated from muscle weight (specific force). This indicates that taurine was not detrimental to the inherent contractile properties of muscle(s). Instead, the loss of force can be attributed to smaller body and muscle weights in taurine treated young mdx mice, relative to age-matched untreated mdx mice.
The decrease in body weight and muscle size was also associated with a decrease in tibia length of taurine treated mdx mice, indicating that the high dose taurine caused growth restriction in these young mdx mice. Whilst studies have shown that dietary taurine restriction can lead to growth restriction [34, 35], evidence that excess taurine can also cause growth restriction is limited in the literature. One study showed that treating newly weaned pigs for 4 weeks with 3% taurine (in food) significantly reduced the weight gain to feed ratio, suggesting some effect of taurine treatment on growth rate, although this did not affect final body weight . Additionally, feeding juvenile sablefish with a range of taurine concentrations (from 0.25% to 6% in food) resulted in growth restriction at low and high concentrations of taurine, with optimal weight gain occurring between 0.4% and 4.2% dietary taurine .
Our data indicate that a high dose of taurine may not be an effective or safe treatment for very young growing mdx mice. In contrast, De Luca and colleagues did not observe any deleterious effects of taurine when treating mdx mice aged 4 weeks (until 8 weeks) with an equivalently high dose of taurine (up to 400 mg per mouse per day, which is comparable with our dose of 16 g/kg/day). Their dose was also highly effective at improving strength of exercised mdx muscles, and was more beneficial than treatment with prednisolone, the current gold standard treatment of DMD boys . There are a number of possible explanations for the discrepancy between our study and that of De Luca. In the De Luca study, mice underwent chronic treadmill exercise, which may have had effects on metabolism or protein turnover that may have ameliorated any toxic effects of high taurine. Additionally, taurine treatment of young mdx mice in the De Luca study began at 4 weeks, while in the current study mice had access to taurine enriched water from 7 days of age. The growth phase of mice is relatively very short, with rapid growth up until 6 weeks before a marked decline of growth until almost ceases by 10–12 weeks . By treating mice from 1–6 weeks of age, we captured a greater portion of their period of maximal growth, whereas mice aged 4–8 weeks in the De Luca study missed the important earlier intense growth phase, with associated age-related changes in metabolism. Regardless of the speculative causes for the discrepancy, results from the current study indicate that taurine treatment regimes can have unforeseen side effects. The usefulness of taurine as a treatment for dystrophy would benefit from further research into the circumstances (e.g. dosing, exercise) in which the side-effects are manifested.
We propose that the detrimental effects of taurine on growth are an indirect effect caused by increased cysteine levels in the liver and plasma. Excess cysteine is considered toxic (reviewed in ) and excess dietary cysteine is known to reduce growth rates . We have previously shown that treating mdx mice with the cysteine precursor OTC from weaning to 6 weeks led to growth restriction  similar to that observed in this current study. In the present study the high dose of taurine increased cysteine levels in mdx liver and plasma and it seems likely that this resulted from a down-regulation of taurine synthesis in the liver, which functions to dispose of excess cysteine .
Cysteine is catabolised by the enzyme cysteine dioxygenase to cysteine sulfinate, which is decarboxylated by the enzyme cysteine sulfinate decarboxylase to form hypotaurine, which is then oxidised to taurine . We found that the high dose of taurine decreased cysteine dioxygenase content and activity, and dramatically decreased cysteine sulfinate decarboxylase content in mdx liver. The activity and levels of these enzymes are highly sensitive to cysteine liver content and dietary intake of cysteine, as the body pool of cysteine requires tight regulation. While cysteine is toxic at high levels, adequate levels are required for protein and glutathione synthesis .
Our observation that taurine exerts a strong feedback effect on this pathway appears to be novel. Although a high protein diet has also been shown to affect activity and levels of these enzymes, this response was suggested to be wholly dependent on the sulfur containing amino acids cysteine and methionine within the protein diet [43–46]. One relevant study showed that cysteine sulfinate decarboxylase was subject to end-product feedback inhibition, since dietary taurine supplementation in healthy rats caused a significant decline in activity . We were not able to locate any studies that suggested that taurine could cause toxicity as a result of excess cysteine. Taurine therapy in adult humans has been reported to be safe and non-toxic, with no adverse effects reported (with the exception of minor gastrointestinal issues) . Our observations are in mdx mice only, and we have previously shown that taurine synthesis is dysregulated in mdx mice  and there appear to be different metabolic demands in dystrophic mdx mice . Such differences may render mdx mice more sensitive to cysteine toxicity (caused by a taurine treatment), compared with other mouse strains; this is supported by research  showing that while taurine treatment (3% in drinking water) decreased weight of adult mdx mice, C57 mice were not affected. Further research is required to establish if high doses of taurine are unsafe in other animal models and humans.
We and others have provided strong data from pre-clinical studies to support the use of taurine as a highly promising and inexpensive therapeutic nutritional intervention for DMD. The issue of growth restriction with high taurine dosage in very young dystrophic animals adds a cautionary note. We propose that this concern can be addressed by carefully monitoring levels of metabolites of taurine and cysteine in the blood and urine of growing animal models and DMD boys. As a consequence, further research is required to identify the dosing range of taurine that maximises the protective benefits of taurine while minimising the likelihood of growth restriction, before considering clinical translation to young DMD boys.
This research was supported by funding from the National Health and Medical Research Council (NHMRC) of Australia.
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- 2. Emery AE. The muscular dystrophies. Lancet. 2002;359: 687–95. pmid:11879882
- 3. Lapidos KA, Kakkar R, McNally EM. The dystrophin glycoprotein complex: signaling strength and integrity for the sarcolemma. Circ Res. 2004;94: 1023–31. pmid:15117830
- 4. Petrof BJ, Shrager JB, Stedman HH, Kelly AM, Sweeney HL. Dystrophin protects the sarcolemma from stresses developed during muscle contraction. Proc Natl Acad Sci U S A. 1993;90: 3710–4. pmid:8475120
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- 8. Shimizu-Motohashi Y, Miyatake S, Komaki H, Takeda S, Aoki Y. Recent advances in innovative therapeutic approaches for Duchenne muscular dystrophy: from discovery to clinical trials. Am J Transl Res. 2016;8: 2471–89. pmid:27398133
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- 10. Matthews E, Brassington R, Kuntzer T, Jichi F, Manzur AY. Corticosteroids for the treatment of Duchenne muscular dystrophy. Cochrane Database Syst Rev. 2016: CD003725. pmid:27149418
- 11. Manzur AY, Kuntzer T, Pike M, Swan A. Glucocorticoid corticosteroids for Duchenne muscular dystrophy. Cochrane Database Syst Rev. 2008;3: CD003725.
- 12. Radley HG, De Luca A, Lynch GS, Grounds MD. Duchenne muscular dystrophy: focus on pharmaceutical and nutritional interventions. Int J Biochem Cell Biol. 2007;39: 469–77. pmid:17137828
- 13. De Luca A. Pre-clinical drug tests in the mdx mouse as a model of dystrophinopathies: an overview. Acta Myol. 2012;31: 40–7. pmid:22655516
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- 15. De Luca A, Pierno S, Liantonio A, Cetrone M, Camerino C, Simonetti S, et al. Alteration of excitation-contraction coupling mechanism in extensor digitorum longus muscle fibres of dystrophic mdx mouse and potential efficacy of taurine. Br J Pharmacol. 2001;132: 1047–54. pmid:11226135
- 16. De Luca A, Pierno S, Liantonio A, Cetrone M, Camerino C, Fraysse B, et al. Enhanced dystrophic progression in mdx mice by exercise and beneficial effects of taurine and insulin-like growth factor-1. J Pharmacol Exp Ther. 2003;304: 453–63. pmid:12490622
- 17. Cozzoli A, Rolland JF, Capogrosso RF, Sblendorio VT, Longo V, Simonetti S, et al. Evaluation of potential synergistic action of a combined treatment with alpha-methyl-prednisolone and taurine on the mdx mouse model of Duchenne muscular dystrophy. Neuropathol Appl Neurobiol. 2011;37: 243–56. pmid:20618838
- 18. Terrill JRP, Grounds MD, Arthur PG. Increased taurine in pre-weaned juvenile mdx mice greatly reduces the acute onset of myofibre necrosis and dystropathology and prevents inflammation. PLoS Curr. 2016;8.
- 19. Terrill JR, Pinniger GJ, Graves JA, Grounds MD, Arthur PG. Increasing taurine intake and taurine synthesis improves skeletal muscle function in the mdx mouse model for Duchenne Muscular Dystrophy. J Physiol. 2016;594: 3095–110. pmid:26659826
- 20. Capogrosso RF, Cozzoli A, Mantuano P, Camerino GM, Massari AM, Sblendorio VT, et al. Assessment of resveratrol, apocynin and taurine on mechanical-metabolic uncoupling and oxidative stress in a mouse model of duchenne muscular dystrophy: A comparison with the gold standard, alpha-methyl prednisolone. Pharmacol Res. 2016;106: 101–13. pmid:26930420
- 21. Horvath DM, Murphy RM, Mollica JP, Hayes A, Goodman CA. The effect of taurine and beta-alanine supplementation on taurine transporter protein and fatigue resistance in skeletal muscle from mdx mice. Amino Acids. 2016: 1–11.
- 22. Marcinkiewicz J, Kontny E. Taurine and inflammatory diseases. Amino Acids. 2014;46: 7–20. pmid:22810731
- 23. Winterbourn CC. Biological reactivity and biomarkers of the neutrophil oxidant, hypochlorous acid. Toxicology. 2002;181–182: 223–7. pmid:12505315
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- 25. Terrill JR, Duong MN, Turner R, Le Guiner C, Boyatzis A, Kettle AJ, et al. Levels of inflammation and oxidative stress, and a role for taurine in dystropathology of the Golden Retriever Muscular Dystrophy dog model for Duchenne Muscular Dystrophy. Redox Biol. 2016;9: 276–86. pmid:27611888
- 26. Terrill JR, Grounds MD, Arthur PG. Taurine deficiency, synthesis and transport in the mdx mouse model for Duchenne Muscular Dystrophy. Int J Biochem Cell Biol. 2015;66: 141–8. pmid:26239309
- 27. Ramsey KA, Bakker AJ, Pinniger GJ. Fiber-type dependence of stretch-induced force enhancement in rat skeletal muscle. Muscle Nerve. 2010;42: 769–77. pmid:20976780
- 28. Hakim CH, Wasala NB, Duan D. Evaluation of muscle function of the extensor digitorum longus muscle ex vivo and tibialis anterior muscle in situ in mice. J Vis Exp. 2013: e50183–e.
- 29. Winterbourn CC, Kettle AJ. Biomarkers of myeloperoxidase-derived hypochlorous acid. Free Radic Biol Med. 2000;29: 403–9. pmid:11020661
- 30. Terrill JR, Boyatzis A, Grounds MD, Arthur PG. Treatment with the cysteine precursor l-2-oxothiazolidine-4-carboxylate (OTC) implicates taurine deficiency in severity of dystropathology in mdx mice. Int J Biochem Cell Biol. 2013;45: 2097–108. pmid:23892094
- 31. Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012;9: 671–5. pmid:22930834
- 32. Murphy RM, Lamb GD. Important considerations for protein analyses using antibody based techniques: down‐sizing Western blotting up‐sizes outcomes. The Journal of physiology. 2013;591: 5823–31. pmid:24127618
- 33. Stipanuk MH, Dominy JE Jr., Ueki I, Hirschberger LL. Measurement of Cysteine Dioxygenase Activity and Protein Abundance. Curr Protoc Toxicol. 2008;38: 6 15 1–6 25.
- 34. Hayes K, Stephan ZF, Sturman JA. Growth depression in taurine-depleted infant monkeys. J Nutr. 1980;110: 2058–64. pmid:7420209
- 35. Hu JM, Rho JY, Suzuki M, Nishihara M, Takahashi M. Effect of taurine in rat milk on the growth of offspring. J Vet Med Sci. 2000;62: 693–8. pmid:10945285
- 36. Liu Y, Mao XB, Yu B, He J, Zheng P, Yu J, et al. Excessive dietary taurine supplementation reduces growth performance, liver and intestinal health of weaned pigs. Livestock Science. 2014;168: 109–19.
- 37. Johnson RB, Kim SK, Watson AM, Barrows FT, Kroeger EL, Nicklason PM, et al. Effects of dietary taurine supplementation on growth, feed efficiency, and nutrient composition of juvenile sablefish (Anoplopoma fimbria) fed plant based feeds. Aquaculture. 2015;445: 79–85.
- 38. Grounds MD, Shavlakadze T. Growing muscle has different sarcolemmal properties from adult muscle: A proposal with scientific and clinical implications Reasons to reassess skeletal muscle molecular dynamics, cellular responses and suitability of experimental models of muscle disorders. Bioessays. 2011;33: 458–68. pmid:21500235
- 39. Stipanuk MH, Dominy JE, Lee JI, Coloso RM. Mammalian cysteine metabolism: New insights into regulation of cysteine metabolism. J Nutr. 2006;136: 1652s–9s. pmid:16702335
- 40. Harper A, Benevenga N. Effect of disproportionate amounts of amino acids. Physiol Rev. 1970;50: 429–539.
- 41. Stipanuk MH. Role of the liver in regulation of body cysteine and taurine levels: a brief review. Neurochem Res. 2004;29: 105–10. pmid:14992268
- 42. Huxtable R. Expanding the Circle 1975–1999: Sulfur Biochemistry and Insights on the Biological Functions of Taurine. Taurine 4. 2002: 1–25.
- 43. Bella DL, Kwon YH, Hirschberger LL, Stipanuk MH. Post-transcriptional regulation of cysteine dioxygenase in rat liver. Adv Exp Med Biol. 2000;483: 71–85. pmid:11787651
- 44. Bagley PJ, Stipanuk MH. Rats Fed a Low-Protein Diet Supplemented with Sulfur Amino-Acids Have Increased Cysteine Dioxygenase Activity and Increased Taurine Production in Hepatocytes. J Nutr. 1995;125: 933–40. pmid:7722697
- 45. Jerkins AA, Steele RD. Dietary sulfur amino acid modulation of cysteine sulfinic acid decarboxylase. Am J Physiol. 1991;261: E551–E5. pmid:1951678
- 46. Jerkins AA, Jones DD, Kohlhepp EA. Cysteine sulfinic acid decarboxylase mRNA abundance decreases in rats fed a high-protein diet. J Nutr. 1998;128: 1890–5. pmid:9808639
- 47. Eppler B, Dawson R Jr. Dietary taurine manipulations in aged male Fischer 344 rat tissue: taurine concentration, taurine biosynthesis, and oxidative markers. Biochem Pharmacol. 2001;62: 29–39. pmid:11377394
- 48. Shao A, Hathcock JN. Risk assessment for the amino acids taurine, L-glutamine and L-arginine. Regul Toxicol Pharmacol. 2008;50: 376–99. pmid:18325648
- 49. Radley-Crabb HG, Marini JC, Sosa HA, Castillo LI, Grounds MD, Fiorotto ML. Dystropathology increases energy expenditure and protein turnover in the mdx mouse model of duchenne muscular dystrophy. PLoS ONE. 2014;9: e89277. pmid:24586653


The Chicago White Sox have released their playoff roster and there are some surprises.
The Chicago White Sox are playing in their first playoff game today in 12 years. It has been a long time since that 2008 defeat to the Tampa Bay Rays but their time to win is back. They have the Oakland Athletics for a short three-game series in the bay. Ahead of their game one, they released their playoff roster and it has some surprises. There are a few players missing that might surprise some fans even though it shouldn’t.
Of course, there are some locks. Lucas Giolito is the ace of the staff and will start game one. Dallas Keuchel is the guy who will take the ball in game two. After that, there are some question marks. Reynaldo Lopez and Gio Gonzalez are off the roster which is a bit surprising. Before being dominated in his last start, there are some people who thought Lopez might get the ball in game three but that isn’t going to happen. Gonzalez hasn’t been good either so we don’t have to worry about him coming out of the bullpen.
Leury Garcia and Zack Collins made their returns to the roster which is something of note. Garcia is going to be in the starting lineup to open the playoffs as Eloy Jimenez is still coming back from an injury. Collins is a good addition because now the White Sox have both James McCann and Yasmani Grandal as designated hitter options. Whoever isn’t catching is the DH and Collins is the backup catcher. It is a sign of good depth here and something fans should be excited about.
This is a roster that the White Sox have been working to build for a long time. Even with a guy like Garrett Crochet who was just drafted there, they have a lot of options that can make this a winnable series. The starting rotation is in doubt after the big two but they have some good bullpen pieces to work with. There are also two MVP candidates in Jose Abreu and Tim Anderson supplemented by players like Yoan Moncada, Luis Robert, and Nick Madrigal so they should be able to score some runs. Get ready because this roster is good and can win some playoff games.


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In a house called Tanglewreck lives a girl called Silver and her guardian Mrs Rockerbye. Unbeknown to Silver there is a family treasure in the form of a seventeenth century watch called The Timekeeper and this treasure holds the key to the mysterious and frightening changes in time. When Silver goes on the run to try and protect herself and The Timekeeper a remarkable and compelling adventure unfolds full of brilliance and wit, as is befitting an author with the imagination and style of Jeanette Winterson.
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'If you want true literary genius, read Jeanette Winterson's young-adult novels, a stunning new facet of her career which began with Tanglewreck. They're too clever for me to summarise' Guardian
About Jeanette Winterson
This is Jeanette Winterson's second book for children. Her first book, King of Capri illustrated by Jane Ray was very well received. Jeanette's first novel, 'Oranges Are Not The Only Fruit' won the Whitbread Award.

domain: library


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Biomedical Science is the term for the investigations, carried out by Biomedical Scientists on samples of tissue and body fluids, to diagnose disease and monitor the treatment of patients. Scientists work in partnership with doctors and other healthcare professionals to perform many different roles in medical laboratories. Biomedical Science is a continually changing dynamic profession and involves study of the diverse areas of medical science including: Biochemistry, Microbiology, Genetics, Haematology and Transfusion Science. It provides training in ‘state of the art’ technologies, to facilitate investigation of disease and medical research.
This direct-entry degree course is offered jointly by University College Cork and Munster Technological University. The degree programme was developed jointly by the two institutes to improve course structure, curriculum, student experience and career opportunities. The programme is managed, administered and taught through both institutions.
There are thirty places available on the programme via CAO code MT871.
This degree programme is accredited and recognised by the Academy of Medical Laboratory Sciences in Ireland. Clinical placement of students in hospital laboratories is required for accreditation.
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Why Choose Biomedical Science at UCC?
The skills and qualification can be transferred all over Europe and are recognised worldwide.

domain: medical


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About Luxury Portable Trailers
Luxury Portable Toilets and Cool Rooms are sub-brands of the Cliff family’s creation – Luxury Portable Trailers. This local family business began with Luxury Portable Toilets in 2015 and has brothers Tim and Brad Cliff at the helm. Providing boutique toilet facilities for special events like weddings, birthdays and anniversaries, the Luxury Portable team pride themselves on excellent facilities and service.
Over recent years, their brand has evolved and grown with full family involvement and the expansion to an all-encompassing brand Luxury Portable Trailers.
Based in Tumbi Umbi on the Central Coast of NSW, Australia. Luxury Portable Toilets and Cool Rooms are available across the Central Coast, North Sydney, Hunter Valley and Newcastle areas.

domain: geography


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NOVEMBER 22, 2017, Mississauga, ON – Results from J.D. Power Canadian Retail Banking Satisfaction Study indicate that while banks have made customer satisfaction strides with mobile banking, other channels, such as the branch, have seen satisfaction decline. To thrive in today’s market, says J.D. Power, banks must adopt a multi-channel approach that promotes consistent customer satisfaction.
Cummins Allison, the leading innovator and provider of coin, currency and cheque processing technology explains how banks can leverage customer-centric technology and outside partnerships to reboot the in-branch experience, increase customer satisfaction and drive product innovation.
Deliver a Better In-Branch Customer Experience & Increase Customer Satisfaction
According to J.D. Power, “banks need to maintain focus on the traditional foundations of the banking experience.” Technology plays a vital role in helping banks deliver an enhanced experience at the traditional branch. For example, when branches shift away from high-volume, time-consuming transactions, they can make banking smoother, faster and simpler in addition to reallocating resources to more consultative services to better support customers.
Self-service coin machines allow branches to automate or eliminate manual coin-handling procedures while processing more coin in less time. Through automation, the branch benefits from increased traffic and improved teller efficiencies, while customers benefit from an improved experience.
Self-service coin counting services fill a unique and powerful role in helping to build customer satisfaction by reducing in-branch wait times, a major cause of frustration. By providing a self-service option, tellers no longer have to spend time in the back-office counting coin since customers can perform this task themselves. And customers have a faster, more meaningful exchange with tellers when they cash-in their coin redemption receipts.
These more meaningful exchanges can have a major impact on overall customer satisfaction. As noted in the J.D. Power study, overall satisfaction scores were 83 points higher among customer who were greeted at the branch entrance and 63 points higher among those whose teller addressed them by name.
Place a Priority on Innovation
Product and service innovation can also impact customer satisfaction. By working closely with vendors that place a high value on R&D and maintain strong patent portfolios, banks put themselves in a position to reap the rewards of their vendors’ innovation and meet the needs of their customers.
Vendors that place an emphasis on financial service innovation feature large teams of engineers that are continually looking to develop the next big breakthrough, while also enhancing current products to offer more features or make them more durable and efficient. By doing so, these partners help banks stay ahead of the market and ensure that solutions developed are in line with the issues being faced by their branches.
As banks rethink their branch strategy, technology investment should be a critical part of the decision-making process. To learn more about you can improve branch productivity and improve customer satisfaction, visit www.cumminsallison.ca/go/automation.
About Cummins Allison
Cummins Allison is the leading innovator and provider of currency, coin and cheque handling solutions.
Our world-class sales and service network includes hundreds of local representatives in more than 50 offices in North America, 6 wholly-owned subsidiaries and is represented in more than 70 countries around the world. For more information about our award-winning solutions, visit www.cumminsallison.ca.
Carol Moore – Vice President Marketing
Outlook Marketing Services, Inc.

domain: economics


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Distressed sales of own-brands and rankings fell across the board From January to May in 2012, automobile production and sales reached 8.003 million units and 802.35 million units, which represented a year-on-year increase of 3.19% and 1.70% respectively. Passenger car production and sales amounted to 6.40700 and 6.33 million, an increase of 7.05% and 5.48%, respectively.
Among them, the sale of self-owned brand cars was 1,193,300, down 8.3% year-on-year, accounting for 27.5% of the total sales of cars, and the occupancy rate was down 3.9 percentage points year-on-year. Compared with the same period of last year, the brands of cars entering the top ten, except Changan Ford Mazda and Chery, declined, and other companies maintained steady growth.
When the growth rate is lower than the average industry data and evolved into a normal state, it is a dangerous signal, and the self-owned brands are not only below the average level, but also have experienced an overall decline, indicating that the crisis has occurred.
The reporter reviewed the sales and ranking changes of self-owned brand sedan from January to May released by the China Association of Automobile Manufacturers from 2010 to 2012. It was found that out of the top ten brands, sales of Chery, Geely, and BYD declined, among which BYD declined most. Seriously, from 253,600 vehicles in 2010, it dropped to 175,300, and Chery also dropped to 173,300 from 195,000 in 2010. From the rankings, except for Geelyâ€™s slight increase, Chery and BYD all declined, and BYD even missed the top 10.
These data all indicate that the market competition pressure of self-owned brand passenger cars is gradually increasing.
Sales/ranking of self-owned brand cars from January to May of 2010-2012 (10,000 units/name)
Car prices 2010 2011 2011 Chery 19.5/7 19.85/6 17.33/10
Geely 17.3/8 18.3/8 17.8/7
BYD 25.36/4 19.65/7 17.53/11
Domestic and foreign joint ventures have jointly squeezed their own "joint autonomy." Such as overnight, they entered the Chinese auto market and formally started a battle against the joint venture brand "Beggar".
Guangzhou Automobile Honda Concept S1, SAIC-GM-Wuling Baojun 630, Dongfeng Nissan Kai Chen D50, Dongfeng Honda Siming, currently have four joint venture models of their own brands to enter, followed by their pace, FAW-Volkswagen, Beijing Hyundai, Changan Ford, etc. The "multinational force" consisting of 10 joint ventures is eager to try.
The market of 80,000 yuan or less has always been the place where independent brands depend on. They have kept their "self-retained land" firmly with high price-to-price ratio. The joint venture car companies that firmly control the high-end market in China and the independent brands have been â€œbehavingâ€ under this market. The emergence of the joint venture's own brand, the car price is approaching 60,000 yuan in the low-end market, direct bayonet see red, the survival of the independent brand is greatly squeezed.
In fact, it is not the prices of joint ventures that are anxious for independent brands, but the control of their brand strengths and product quality. At present, China's own brands have a large gap between the core technologies and foreign technologies in engines, transmissions, and chassis. At present, independent brands do not have the brand appeal. This is also doomed to take the high road and is full of twists and turns.
In recent years, Geely, Chery, Aeolus and other independent brands have made attempts to enter the high-end car market. However, there are not many successes. Only the Emgrand EC7, BYD S6, and Haval H6 have won recognition from the market.
In addition, a series of auto consumption incentives, such as a halve of purchase tax, automobile out of the countryside, trade-in replacement, and so on, will undoubtedly further exacerbate the sales pressure of independent brands this year.
What are the most important needs of a self-owned brand to promote a boutique model SUV? A good product. Although the current independent brands also have sales of tens of thousands of models, but really can be said to have a reputation for products, almost one of the few. Under internal and external problems, every product that is refined is gradually valued by them.
This idea of â€‹â€‹promoting quality consciousness emerged in 2011. BYD S6, Chery E5, China H530, Great Wall Tengyi C50, SAIC MG3, SAIC Roewe W5 and other models, have made significant progress. This year, self-owned brand models will be upgraded. Such as Chang'an Yidong, Fengshen A60, FAW Oulang, Lifan new 520, the new Roewe 750 and many other models will be listed. In addition, the SUV market is still gaining momentum in the current downturn in the market. Naturally, independent brands do not miss this opportunity. Including Roewe W5, Emgrand EX7, BYD S6, Huatai B35, Haval H6, Lifan SUV, King Yi SUV and other quietly listed, which also provides a new world for independent brands.
New Energy Vehicles Curve Overtaking With the continuous rise in oil prices and peopleâ€™s attention to air pollution emissions, peopleâ€™s awareness and acceptance of new energy vehicles have greatly increased.
At present, new energy vehicles have become "hot" in China. From the country's formulation of strategic emerging industries planning, to the local government to vigorously build new energy vehicle bases, and then to the car enterprises to upgrade new energy vehicles to a strategic height, new energy vehicles are entering the critical period of industrial development.
The â€œEnergy Conservation and New Energy Vehicle Industry Development Plan (2012-2020)â€ was approved by the State Council, which clarified the main strategic orientation of the transformation of pure electric vehicles into the automobile industry, promoted the industrialization of pure electric vehicles, and promoted the overall technical level of the domestic automotive industry. . For the Chinese auto industry, new energy vehicles are likely to become a powerful starting point for transforming development methods, implementing their own brand strategy, or even turning from strong to strong. After all, under the background of the traditional internal combustion engine vehicles being controlled by people, by vigorously developing new energy vehicles characterized by â€œenergy conservation and environmental protectionâ€, it is not only in line with the development trend of the worldâ€™s automotive industry, but also in line with the direction of domestic auto companies to enhance their independent innovation capabilities.
In recent years, domestic brands have begun to â€œbig moneyâ€ and their brand image has greatly improved.
The marriage between Geely and Volvo created the first move. Currently. Geely and Volvo also reached an agreement to transfer technology. The two sides also agreed to establish a joint venture company to achieve Volvo domestic production. The two parties stated that they must jointly develop small-displacement, high-performance, green-environmental series engines, environmentally-friendly compact car platforms and new energy vehicle assembly system technologies.
Chery and Land Rover jointly announced that they will invest 17.5 billion yuan to establish a joint venture company in China to produce whole vehicles and engines, and will launch a joint venture own brand. The electric vehicles jointly produced by BYD and Daimler also appeared at the Beijing Auto Show. The car was led by the Daimler design team to shape, body and chassis. BYD was responsible for the development of the electric drive system.
At this point, the three domestic automotive companies, most of whom are expected by consumers to develop their own brands, have chosen to rely on foreign brands to achieve technological, brand, and even profit enhancements. In addition, Great Wall Motor once discussed the joint venture with Jaguar Land Rover. Jianghuai Automobile had also been in contact with Subaru Motors. Young cars have been trying to acquire Saab. Through joint ventures with foreign car companies to upgrade brand technology, it is no longer a patent of FAW, Second Automobile, SAIC, BAIC and other large state-owned enterprises.
In the face of huge market pressure, some independent brands began to adjust their expansion pace and started to pay attention to the quality of distributors. It is reported that based on the existing network of more than 600 dealers, Chery Automobile will no longer add new dealers this year, and will instead support the development of existing dealers, improve service quality and improve profitability.
At present, the most healthy self-developed brand in the market is Great Wall Motors. In the Guangzhou market, there is currently only a large distributor in Guangdong Youdao. This is a monopoly operation, which gives dealers control of market segments and guarantees profit to support them well.
This year, Great Wall Motors will also focus on improving the distribution structure. It will eliminate 20% of dealers' outlets that are not up to standard to improve service quality. According to Great Wall Motor's calculation in 800 domestic distribution outlets, eliminating 20% â€‹â€‹of its outlets means that 160 dealers will be liquidated during the year.
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Ashkahn "Thank you" with illustrated handshake. Sexy letterpress on lux cotton paper. Blank on the inside. Comes with Ashkahn trademark fluorescent pink envelope.
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Source: Deep Learning on Medium
Clearing the Confusion using Examples
We all have been caught in the confusion of differentiating between Artificial Intelligence (AI) vs Machine Learning (ML) vs Deep Learning (DL). Haven’t we?
Although the terminologies are always used interchangeably, there do not quite refer to the same things.
This article will give you a very clear understanding of these terms with specific examples so that you can understand which is better for your specific use case: Artificial Intelligence, Machine Learning, or Deep Learning.
What is Artificial Intelligence (AI)?
Artificial Intelligence (AI) is all about incorporating human intelligence into machines. It gives the ability to machines to recognize human’s face, move and manipulate objects, understand the voice of human’s, or solve other tasks.
An Artificial Intelligent (AI) machine can see its surrounding (using a camera), listen to others (using a microphone), touch objects (using robotic arms), speak to others(using a speaker) and do all these intelligently just like human’s do.
Example’s of Artificial Intelligence (AI):
- Self Driving Cars: Artificial Intelligence (AI) can be used to create a fully autonomous car’s to travel between destinations without a human operator.
2. Sign and Voice Recognizer (SVR): SVR is a part of a hackathon project me and my roommate, Mohit, build during our Internship at Microsoft, India. SVR (pronounced as स्वर, sʋəɾ) is a glove that is aimed at helping people with impaired hearing and people who have difficulty in speaking/ talking to others: It recognizes sign language gestures of the wearer and converts them to speech.
3. Sophia — The Humanoid Robot: Sophia is a social humanoid robot developed by Hong Kong-based company Hanson Robotics. Sophia is the first humanoid robot which is being granted a ‘robot citizenship’ by Saudi Arabia.
After seeing all these wonderful examples, one question arises, “What algorithms give these robots, or cars, or gloves the capability to think and make decisions intelligently?” The answer is Machine Learning (ML).
What is Machine Learning (ML)?
Machine Learning (ML) is a subset of Artificial Intelligence (AI) technique which use statistical methods to enable machines to improve with experience. Or, if put simply,
Machine Learning empower’s computer systems with the ability to learn.
The intentions of Machine Learning is to enable machines to learn by themselves using the provided data and make accurate predictions.
Example of Machine Learning (ML):
- Snapchat Filters: Snapchats filters use augmented reality and machine learning for your flower crowns selfies.
2. Netflix — The Application that knows what you want: The Netflix recommendation system is powered by Machine Learning (ML) algorithms and that is what makes their system so good in predicting the movies that you are most likely to watch and hence show them as suggestions to you.
Machine Learning (ML) algorithms give the ability to the Artificial Intelligence (AI) machines (either Sophia robot, or cars, or SVR glove) the ability to learn from data and make intelligent predictions which AI machines use as their decisions. So, in short, you can think of Machine Learning (ML) as the technique of Artificial Intelligence (AI) which gives them the ability to learn and act according to data.
“Can Machine Learning (ML) algorithms be made so much efficient that they outperform the human performance at a particular task?” The answer is yes, and that is what Deep Learning (DL) algorithms are used for.
What is Deep Learning (DL)?
Deep Learning (DL) is a particular kind of Machine Learning (ML) algorithm that is inspired by the functionality of our brain cells called neurons which led to the concept of Artificial Neural Network (ANN).
Deep Learning (DL) is what is behind the most human-like Artificial Intelligence (AI).
Examples of Deep Learning:
- Language Translation — Google Translate: Translating from one language (say English) to another language (say Spanish) is now just a click away, thanks to Deep Learning algorithms.
2. Gmail Smart Compose and Google Photos Color Pop: Gmail’s ‘Smart Compose” feature will suggest small, brief responses to your emails based on their content. Google Photos, on the other hand, will allow users to automatically color relevant black and white photos, automatically detecting the subject’s photo and coloring their skin, clothing, and more. Deep Learning algorithms are used to make these features so great.
Reading all these, one question arises, “Why do we even need to study Machine Learning when Deep Learning is so good?”
This question is very relevant and most of the big tech companies (like Google, Microsoft, Amazon, Facebook) are implementing deep learning models to achieve better performance and enhancing user experience.
However, Deep Learning (DL) algorithms require:
- A huge amount of data as compared to other Machine Learning (ML) algorithms to learn and predict intelligently.
- A Huge computation power to train like GPUs (which are very expensive)
- More specialization and expertise.
However, don’t let this to stop you from learning and implementing deep learning algorithms as there are multiple free solutions to these requirements. (Google Colab — offers Nvidia Tesla K80 GPU (which cost around 4000 dollars) and 12 GB of RAM from free).
If we have to summarise this to one sentence, that would be “Artificial Intelligence (AI) is the all-encompassing concept that initially erupted, then followed by Machine Learning (ML) that thrived later, and lastly Deep Learning (DL) that is promising to escalate the advances of Artificial Intelligence (AI) to another level.”
In the end, I will leave you with this very famous image used to distinguish AI vs ML vs DL. As you can see on the above image, Deep Learning (DL) is a subset of Machine Learning (ML), which is also a subset of Artificial Intelligence (AI).
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Meet Victoria Harwood Butler-Sloss
Victoria Harwood Butler-Sloss is half Armenian, half English, brought up in Nicosia, Cyprus. She moved to London at eighteen and began her career as a dancer at the Raymond Revuebar, She trained at RADA, the Royal Academy of Dramatic Art, and spent twenty years as an actress, playing a slew of mad, bad, exotic foreigners on British television and performing in the West End, The Royal Court and European tours amongst others. After moving to Los Angeles, Victoria wrote her first book, The Seamstress of Ourfa, the first in a trilogy beginning in the Ottoman Empire in 1895 and following four generations of women until the present day.
“A Flock of Birds” was her first film as writer/director/producer/DP. Based on a chapter from her book it is based on a family story with a fictional twist. Previously she produced, wrote and appeared in “Cyprus Summer 1974” a short documentary based on a diary she wrote during the coup and subsequent war that divided Cyprus, her home.
She is currently working on another project, “Rest in Pieces,” a docudrama about death and what it means to be buried in a cemetery in no man’s land, awarded a Spark grant to help completion by Creative Armenia Network.
All her work can be seen on her website at planetvix.com.
Appearances at Live On Green


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Pennsylvania State Troopers in riot gear deploy near the intersection of North Avenue and Pennsylvania Avenue May 1, 2015 in Baltimore, Maryland.
BALTIMORE - Six Baltimore police officers will face criminal charges, including second-degree murder and manslaughter, in the death of a black man who was arrested and suffered fatal injuries while riding in a moving police van, the city&39;s chief prosecutor said on Friday.
Marilyn Mosby, the state&39;s attorney for Baltimore City, said Freddie Gray, who died a week after his April 12 arrest, suffered a critical neck injury as a result of being handcuffed inside the van.
Breaking news. FreddieGray was illegally arrested, death deemed a homicide, occurring while he was in police custody.— Talib Kweli Greene (@TalibKweli) May 1, 2015
She said officers failed to provide medical attention to Gray even though he asked for it on at least two occasions.
Warrants have been issued for the arrest of the officers charged in the case, she said. In addition to murder and manslaughter, charges include assault, misconduct and false imprisonment.
In a dramatic news conference, Mosby said the Maryland chief medical examiner ruled Gray&39;s death a homicide and that his spinal injuries occurred while he was riding without restraints in the van.
Gray was no longer breathing when he was finally removed from the van, Mosby told a crowd, which broke into applause after she finished speaking.
Gray&39;s death on April 19, a week after his arrest, is the latest flashpoint in a national outcry over the treatment of African-Americans and other minority groups by US law enforcement.
After a night of rioting in Baltimore on Monday, protests spread to other major cities in a reprise of demonstrations last year set off by police killings of unarmed black men in Ferguson, Missouri, New York and elsewhere.
Without protestors in the streets, I am certain that these 6 officers would still be on the streets. FreddieGray— deray mckesson (@deray) May 1, 2015

domain: news


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Fleetwood Mac Helps Raise $7 Million For CharityRock 'n' roll's dysfunctional family, Fleetwood Mac, joined with artists paying tribute to their work to raise $7 million for down-on-their luck musicians at a benefit in Radio City Music Hall on Friday.
Bob Dylan MusiCares Person of the Year Concert Adds Bruce Springsteen and MorePreviously honored by Musicares have been Tony Bennett, Bono, Natalie Cole, Phil Collins, Neil Diamond and Aretha Franklin.
Ben Harper and Tim McGraw Talk GRAMMY MusiCares Springsteen Tribute
2012 GRAMMY Nominees Album To Be Released January 24The 2012 GRAMMY Nominees album will be released digitally and in stores January 24, 2012. The album features 22 hits from some of the GRAMMYs' most popular nominees.


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For daily tips, please visit our Facebook Page (thank you)
Friday, March 12
Exercise IS medicine. Every year there are 250,000 deaths due to lack of exercise. Diseases like high blood pressure & diabetes both can improve with exercise and a healthy diet! Exercise adds up . . if you can only do 10 min it's 10 min more that you did do!
Thursday, March 11
Blood Pressure . . do you know your numbers? 1 in 3 adults have high blood pressure. It is very hard on your body to have it and NOT treat it. Many folks do not like taking daily medication but the benefits are undeniable! Next time you are waiting at the pharmacy, check your BP and know your numbers.
Wednesday, March 10
Struggling to keep proper form doing a push-up? Did you know you can do them against the wall!? Great way to get started. Place hands at shoulder height and width. Walk back so your heels are just off the ground. Keeping your body straight "fall" into the wall and push away. This is a wonderful way to get started!
Tuesday, March 9
Tree Pose. Improve your balance by standing on one foot and placing the sole of the other foot on the inside thigh of the standing leg. Find your balance. Hold on if yoiu need to, if you keep practicing you won't need to for long! Switch sides.
Monday, March 8
Protect your nugget! Okay you get ONE brain. There are sports specific helmets for a reason. . use them. Helmets are very important for skiingand snowboarding, and practicing safety will extend your snow season.
Friday, March 5
Going for that check-up? Take time before your visit to write down all of the vitamins, and supplements you are taking. Many interfere or affect medication. If your doctor is not sure, consult your local pharmacist. Consults at the pharmacy are free and these are very knowledgeable professionals
Thursday, March 4
H2O to go! Take water with you. Carry a bottle at all times, it will help you to not get a soda and you can meet your daily requirements a little at a time.
Wednesday, March 3
Smiling at someone new today will be good for their heart, or yours.
Tuesday, March 2
Balance. . .use it or lose it! Check your balance . . stand on one foot, see how long you can stand before your topple; when you get that doing better, try closing your eye while on one foot. The point is you have to challenge your balance and not take it for granted, plus it will help reduce falls as you age.
Monday, March 1
Resist snacking after dinner by immediately brushing your teeth! You will be less likely to to want to repeat the process and it will help you wind down for the night.
Friday, Feb 26
Add spice to your life! Getting family to eat veggies can be a good challenge. Try using salsa or a dip as a topping. If you dip 'em do not double dip give each person their own small dipping container. Sometimes just adding a dash of a spice helps too!
Thursdays, Feb 25
Going to the market tonight? Get some fruit (or dried fruit) for that mid-day snack craving. It will keep you away from the vending machine and you will get the benefits of the antioxidants and fiber too!
Wednesday, Feb 24
It's all about the color! When you build your dinner plate (or any meal) think of it like a paint palette, the more color the better! Foods with strong colors have more antioxidants!
Tuesday, Feb 23
Your world starts from the ground up . . invest in GOOD shoes for your physical activity. When you purchase athletic shoes you do not ever need to "break them in" if they are stiff or not perfect, try another brand. Be specific in the type of shoe with the activity. I know the good ones cost a bit more but it's your feet and they are worth it.
Monday, Feb 22
Eating before a workout? If you want to - keep it light. Try to stick to 200 kcals or less. You will burn the MOST kcals up to 2 hours after you are done exercising. My favorite post-exercise snack, chocolate milk! A nice pairing of carbs & protein for the maximum caloric usage.
Friday, Feb 19
Portion Distortion! Do you know what a portion of protien (meat type) should look like?
Thursday, Feb 18
Stress getting you? Traffic? Try turning off the tv and any other outside sound. Take a slow deep breath. If you are not driving put your hands on your waist and when you breathe in feel the expansion at your sides. Exhale through your nose to a slow count of 5. This will help you relax and is sometimes as good as a longer time out.
Wednesday, Feb 17
Eat Less, Chew More! Sit Less, Move More! Frown Less & Smile More!
Tuesday, Feb 16
Trying to fit it in? Cannot stay committed to your workouts? Try setting the same time aside EVERY day and do something. Can be 5 min of jumping jacks, abs or even going for a good brisk walk for 20 min. Remember! Exercise adds up like spare change, get it started and stick with it . . and for more fun, invite a friend!
Thursday, Feb, 11, 2010
A little bit every day helps! Exercise adds up like spare change. Do something every day no matter how big or little, it all counts!
Wednesday, Feb 10, 2010
Keeping your children active will not only improve their life, their lifestyle but also helps to build self-esteem, teamwork and they will focus better in school.
Tuesday, Feb 9, 2010
Fit It In! Why take the elevator when you can walk the stairs? Try 2 at a time. Great for your legs and bum. To be safe, when going down stick with one at a time.
Monday, Feb 8, 2010
If you get a cough and still have to work or be out in public, make sure to cough into your elbow! Spare yourself and others from the germs.
Friday, Feb 5, 2010
Trouble sleeping? If you find yourself counting sheep at night and are still awake after 30 minutes, get up and try some yoga postures. If you are not sure what to do try some meditation or even listening to some soothing tunes. Try to clear your mind and not solve tomorrow's or today's problems.
Thursday, Feb 4, 2010
Need an ice pack quick? Grab that cold soda out of the frig or from your lunch box. Works great when you have no access to ice.
Wednesday, Feb 3, 2010
Inner thighs weak? Try seated in a chair with a sports ball or 4-square ball between your knees. Sit up straight and tall and squeeze slowly for a set of 12, then a quick set of 12. Repeat this twice.
Tuesday, Feb 2, 2010
Runners & Joggers - do you have trouble relaxing your hands while you are running or moving? Try holding an old cadboard toilet paper roll in your hands while you run. You will crush it if you squeeze too hard and you will eventually get to a loose grip, therefore relaxing your hands.
Monday, Feb 1, 2010
Meatless Monday - swap today's burger for a grilled portabella mushroom (30 kcals). Pile on a tomato and some greens and enjoy a meatless substitute for just one day! It's not just a savings on grease and oils . . but also on calories!
Friday, Jan 29, 2010
Scrambled? Over Easy? No matter how you like them eggs can be a wonderful source of protein and vitamins A, B, D & E. Use proper safety when handling the eggs raw making sure to wash your hand after handling them.
Thursday, Jan 28, 2010
Is Sugar Free better? Just because it sounds healthy, does not mean it is! By all means if you are diabetic go for sugar-free, but read the labels. Often the fats/oils and things like sodium are increased a LOT to improve the taste after removing the sugar.
Wednesday, Jan 27, 2010
Blood Pressure - do you know what yours is and why high blood pressure is called the "silent killer"? Know your numbers! Many of us walk around without knowing our blood pressure is high. This can cause great stress and even damage your bood vessels and organs. Those machines in the pharamcy DO work and are pretty accu...rate. Better to treat than go around with this condition!
Tuesday, Jan 26, 2010
Planning saves calories? If you put some nuts, fruit (dried or fresh), or even a sports bar you will be prepared for that mid-morning snack or that mid-day low . . or both! This will not only save you money but kcals from getting something from a vending machine or fast food. Keep it healthy with a little planning, you will have better energy and nutrition.
Monday, Jan 25, 2010
Morning, noon or night- need a quick pick me up? Head to a glass of low-sodium veggie juice! Good calories, long lasting energy and it will satisfy your hunger pangs.
Friday, Jan 22, 2010
Great Grapefruit! Side by side both the pink and white grapefruit both have the same amount of calories and vitamin C. The pink grapefruit have more beta carotene, almost 40x more!
Thursday, Jan 21, 2010
Helmets are not just for kids. Reduce your chance of a brain or head injury by 85%. Protect your "egg" (brain) when biking, snowboarding, skiing, rollerblading . . etc.
Wednesday, Jan 20, 2010
Peanut Butter - although a wonderful source of protein has a lot of fat. One serving size is 2 tablespoons which is the size of two (2) ping pong balls. Don't go for portion distortion!
Tuesday, Jan 19, 2010
Morning Fitness is a great way to not only jump start you day, improve your energy but also fires up your metabolism!
Friday, Jan 15, 2010
Quick Switch! When snacking, try using your less dominant hand. You will actually eat less snacks this way!
Thurs, Jan 14, 2010
Supersize you? If you do not want to supersize your body, make a promise TODAY that you will not supersize your fast food meals - ever. This will help reduce fat and caloric intake.
Wed, Jan 13, 2010
Are you nuts? Walnuts may help to reduce cholesterol - especially the "bad" LDL that is most like to do damage to coronary arteries. Keep control of your portions. Nuts are high in protein but can also be high in calories.
Tues, Jan 12, 2010
Feeling stressed in the new year? Give yourself 30 minutes daily of something fun that you enjoy doing (i.e. take a walk, talk to an old friend, play with your pet, read a good book, watch tv, visit a friend) . . something that is just for you to "take you away" for 30 minutes. When you are done you will find it easier to think positive thoughts and get through the rest of the day.
Mon, Jan 11, 2010
Winter Squash are available NOW in your local grocery store or farmer's markets. They are packed with Vitamins C & A, potassium, dietary fiber, and manganese too! They are loaded with a good source of folate, omega-3 fatty acids, thiamin, copper, vitamin B5, vitamin B6. Easy to cook and tasty too. Time to get 'squashed"!
Wed, Dec 23, 2009
Afraid of the Flu (me 2!) To avoid getting sick in this cold weather time of year, wash your hands! Sing the alphabet song . . a friend tells me he tries to sing it backwards! And remember it is not just the soap and hot water helping but also how you wash and make sure you don't miss a spot.
Tues, Dec 22, 2009
Interesting Intervals! Add cardio intervals (I call them cardio bursts) into your cardio routine or into your strength training program. Althernate short, moderate intensity bursts of cardio into your longer cardio sessions, or between every 2 or 3 strength exercises. Keeps in interesting and fun AND you burn more calories over all!
Mon, Dec 21, 2009
Celebrate the Season! For a different version of your candy jar - try dried fruit and nuts. I prefer dried cranberries and almonds. other like walnuts . . there is a variety of dried fruits available mix it up, make it festive AND healthy!
Fri, Dec 18, 2009
Grapefruits and Medications. Did you know that with some medications too much intake of grapefruit will interfere. Consult with your physician or pharmacist to see if the could be a risk with eating grapefruit.
Thurs, Dec 17, 2009
For my meat eater FB friends . . trim the fat from your meat before you eat. This can cut your fat intake by as much as 50%! So take a moment and cut it off! You don't need it and it will only hurt you.
Wed, Dec 16, 2009
Having trouble getting your workout in first thing in the morning? Try setting your workout clothes by your bathroom or somewhere you can see them when you first wake up. Not only will it be easier to get going they are a silent reminder to take care of yourself.
Tues, Dec 15, 2009Snack Control! When you get home from the grocery store with a bag of chips or cookies or whatever your fav snack is - bag 'em up! Split the snacks into portion sized bags. That way when you have a snack attack, you can just pull the correct portion size and not be tempted by the whole bag.
Building: PE 6

domain: health


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6324 M - FAG Deep Groove Bearing - Quality Bearings Onli
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iPad Air 10.9 Inch Wi-Fi 64GB
Price includes delivery
This product features:
- 10.9-inch Liquid Retina display1 with True Tone, P3 wide colour and an antireflective coating
- Apple M1 chip with Neural Engine
- 12MP Wide camera
- 12MP Ultra Wide front camera with Centre Stage
- Up to 256GB of storage3
- Available in Blue, Purple, Pink, Starlight and Space Grey
- Stereo landscape speakers
- Touch ID for secure authentication and Apple Pay
- All-day battery life4
- Ultrafast Wi-Fi6
- USB-C connector for charging and accessories
- Works with Magic Keyboard, Smart Keyboard Folio and Apple Pencil (2nd generation)2
- iPadOS 15 is uniquely powerful, easy to use and designed for the versatility of iPad
*Apps are available on the App Store. Title availability is subject to change.
1The display has rounded corners. When measured diagonally as a rectangle, the iPad Air screen is 10.86 inches. Actual viewable area is less.
2Accessories are sold separately. Compatibility varies by generation.
3Available space is less and varies due to many factors. A standard configuration uses approximately 12GB to 17GB of space, including iPadOS 15 with its latest features and Apple apps that can be deleted. Apple apps that can be deleted use about 4.5GB of space, and you can reinstall them from the App Store. Storage capacity subject to change based on software version, settings and iPad model.
4Battery life varies by use and configuration; see apple.com/au/batteries for more information.
|iPad Air 10.9 Inch
|Wi-Fi and Bluetooth
|Up to 10 hours of surfing the web on Wi‐Fi or watching video
|802.11ax Wi-Fi 6; simultaneous dual band (2.4GHz and 5GHz); HT80 with MIMO
|Charging via USB-C to a power adapter or computer system
|Magnetic Smart USB-C
|Built‐in 28.6‐watt‐hour rechargeable lithium‑polymer battery
|W 178.5 x H 247.6 x D 6.1 mm
|USB‑C Charge Cable (1 metre)
20W USB‑C Power Adapter


Campbell’s Cuts 1.5 Million Gallons of Water Per Day
In Campbell’s latest corporate sustainability report (PDF) the company says it has reduced water consumption by 1.5 million gallons per day, due to installation of a system which recovers heat and recycles cooling water. The company plans to install similar systems in other facilities.
Based on goals set in 2005, the company has also achieved seven percent energy reduction in North America through initiatives such as energy audits, heat recovery projects and leak-detection techniques. Almost meeting its goal for reducing 10 percent energy-consumption by 2010.
As for recycling, the company reported it has recycled 70 percent of the 157,000 metric tons of solid waste generated by its U.S. manufacturing facilities.
The company is also spearheading the redevelopment of its hometown, Camden, N.J., at an estimated cost of $73 million to the company.
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