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Down syndrome (DS) neurons are more susceptible to oxidative stress and previous studies have shown that vitamin E was able to reduce oxidative stress and improve DS neurons' viability. Therefore, this study was done to investigate the protective role of γ-tocotrienol (γT3) in DS neurons from hydrogen peroxide (H2O2) -induced oxidative stress. The pro-apoptosis tendency of γT3 was compared to α-tocopherol (αT) in non-stress condition as well.
Primary culture of DS and euploid neurons were divided into six groups of treatment: control, H2O2, γT3 pre-treatment with H2O2, γT3 only, αT pre-treatment with H2O2 and αT only. The treatments were assessed by MTS assay and apoptosis assay by single-stranded DNA (ssDNA) apoptosis ELISA assay, Hoechst and Neu-N immunofluorescence staining. The cellular uptake of γT3 and αT was determined by HPLC while protein expressions were determined by Western blot. Comparison between groups was made by the Student's t test, one-way ANOVA and Bonferroni adjustment as well as two-way ANOVA for multiple comparisons.
One day incubation of γT3 was able to reduced apoptosis of DS neurons by 10%, however γT3 was cytotoxic at longer incubation period (14 days) and at concentrations ≥ 100 μM. Pre-treatment of αT and γT3 only attenuate apoptosis and increase cell viability in H2O2-treated DS and euploid neurons by 10% in which the effects were minimal to maintain most of the DS cells' morphology. γT3 act as a free radical scavenger by reducing ROS generated by H2O2. In untreated controls, DS neurons showed lower Bcl-2/Bax ratio and p53 expression compared to normal neurons, while cPKC and PKC-δ expressions were higher in DS neurons. On the other hand, pre-treatment of γT3 in H2O2-treated DS neurons have reduced Bcl-2/Bax ratio, which was not shown in euploid neurons. This suggests that pre-treatment of γT3 did not promote DS cell survival. Meanwhile γT3 and αT treatments without H2O2 as well as pre-treatment of γT3 and αT induced changes in cPKC and PKC-δ expression in DS neurons suggesting interaction of γT3 and αT with PKC activity.
Our study suggests that γT3 pre-treatment are not sufficient to protect DS neurons from H2O2-induced oxidative assault, instead induced the apoptosis process.
Keywords:Apoptosis; Down syndrome; human neurons; oxidative stress; γ-tocotrienol; vitamin E
Vitamin E is a generic term for lipid-soluble, chain breaking antioxidants which consists of four tocopherol isomers (α, β, γ, δ) and four tocotrienol isomers (α, β, γ, δ). The tocopherol and tocotrienol isomers differ in the number and position of methyl substitutions on the chromanol head. Although tocopherols and tocotrienols are closely related chemically, they differ in their biological effectiveness [1]. Studies have shown that vitamin E deficiency impairs cognitive performance in mice subjected to oxidative stress [2]. Meanwhile, one study found that Down syndrome (DS) children have significantly less vitamin E levels than normal children [3]; while another study showed that DS patients with dementia have lower plasma levels of vitamin E than controls without DS [4]. These results suggest that intake of essential nutrients such as folate, vitamin B6, vitamin E, selenium, α-lipoic acid might be important in preventing cognitive deterioration in DS and Alzheimer disease (AD) [5].
However, intervention studies of antioxidant supplementation in DS and AD have not been conclusive. A recent randomized controlled trial on antioxidant supplementation, including vitamin E for DS children did not show any significant difference in developmental outcome after a two-year research period. There was also no significant effect of antioxidant supplementation on the superoxide dismutase and glutathione peroxidase activities, on the superoxide dismutase to glutathione peroxidase ratio and on the urinary isoprostane concentrations [6]. Another recent review that looked at five different studies on antioxidants and cognitive functions revealed that only three studies examining vitamin E and C supplements gave significantly different results-i.e. one study found a positive association with specific cognitive test, while the other two studies showed a link with global cognitive functions [7]. Other double-blind studies reported that vitamin E has no benefit in patients with mild cognitive impairment and Alzheimer's disease [8]. In all these trials, subjects partake high doses of vitamin E (2000 IU or 1500 mg) daily, which is more than the upper tolerable intake level for vitamin E (1500 IU or 1000 mg per day) [9].
Vitamin E mainly function as free radical scavenger, but recent studies showed that tocopherols and tocotrienols have other non-antioxidant roles: α-tocopherol (αT) was shown to modulate signal transduction and gene expression in various cell lines, while tocotrienols possess powerful neuroprotective, anti-inflammatory anti-angiogenic, anti-artherogenic, anti-cancer and cholesterol lowering properties (for a comprehensive review, refer [10]). Vitamin E has been shown to be neuroprotective in various studies: firstly in a landmark study of neurodegeneration of in vitro culture of DS neurons [11]; followed by a study that reported that αT was able to attenuate oxidative stress-induced apoptosis in striatal neuron cultures via its free radical scavenger function [12]; while other studies showed that α-tocotrienol protects neurons from glutamate-induced cell death by the c-src activation molecular pathway [13]. However, not many studies have address the possible pro-apoptotic tendency of vitamin E in neurons, especially tocotrienols, which has shown to have greater apoptotic activity towards various cancer cell lines such as mammary tumor cells and prostate tumor cells compared to tocopherols [14,15]. Current studies has shown that tocopherol and vitamin E analogues were able to induced apoptosis in murine C6 glioma cell line and Tet21N neuroblastoma cell line [16,17] but not tocotrienols.
γ-Tocotrienol (γT3) was reported to activate the apoptosis pathway via the mitochondrial death pathway of the Bcl-2 family proteins in pancreatic stellate cells [18], while tocotrienols was shown to be anti-proliferative in mammary epithelial cells by reducing PKCα (Protein Kinase C) activation [19]. Our previous studies in primary rat's astrocytes and cerebellar neuron cultures revealed that high dosage of γT3 was cytotoxic and have a high tendency to induce the expressions of proteins that were involved in the apoptosis pathway such as Bax, p53 and p38 MAPK [20,21]. Another study also showed that high doses of vitamin E and vitamin C enhanced the toxic effect of H2O2 to cells [22]. Since most trials of vitamin E supplementations utilized high dosage of vitamin E for maximum effects, the concern for the safety of vitamin E supplementation at the molecular and cellular level has yet to be fully addressed. DS cells are known to be highly susceptible to oxidative damage compared to normal cells [23]. Genomic and functional profiling of DS neural progenitor cell line exposed to S100B suggested that dysregulation of chromosome 21 genes led to increased ROS and thereby altered transcriptional regulation of cytoprotective genes in response to oxidative stress [24]. Vitamin E treatment induced neuroinflammatory processes by increasing microglial activation in animals overexpressing S100B, which is involved in the neuropathology of DS and AD [25]. Therefore, the purpose of this study is to further investigate the effects of αT and γT3 in the apoptosis signaling pathway of human DS neurons as a model of oxidative stress susceptible system, while normal human neurons were used as control.
Materials and methods
The Malaysia Palm Oil Board (MPOB) supplied the palm γT3 and αT isomers of 87% and 80% purity respectively, which was isolated as described previously [26]. Culture dishes were from Nunc while antibodies (p53, Bax, Bcl-2, cPKC, PKC-δ, β-actin) were from Santa Cruz Technologies. Reagents for 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were from Promega while single-stranded DNA (ssDNA) Apoptosis ELISA kits were from Chemicon. All other chemicals and reagents were from Sigma unless indicated.
Primary cortical neuron cultures from normal and DS fetal brain
The cultures were established using human cortical brain tissues obtained from normal euploid and DS legally aborted fetuses at 14-21 weeks of gestation. The permission to use human fetal tissues was obtained from the ethics committee of the Spanish National Research Council (CSIC) (Approval date: August 6th, 2009; ref. no: SAF2009-13093-C02-02). Enriched neuron cultures were prepared as described elsewhere [27].
Cell culture treatments
Neuron cultures were incubated with varying concentrations of γT3 and αT, with αT as a positive control based on previous studies showing αT having non-toxic and neuroprotective effects on neurons [12,21]. Stock solutions of 0.5 M γT3 and αT (in 100% ethanol) were first resolved overnight in fetal calf serum at 37°C and diluted to 100 times the final concentration with culture media containing 50% ethanol. Final dilution of αT and γT3 in the cell culture contained 0.5% ethanol, which did not significantly affect cell survival (data not shown). All experiments utilized freshly prepared dilutions of H2O2, αT and γT3.
Cytotoxicity of γT3
The human DS neuron cultures were incubated with γT3 (1-200 μM) for 24 hours at 37°C. DS neurons were also given γT3 treatment of 7 days and 14 days to determine possible cytotoxicity or protective effects of γT3. Cytotoxicity of γT3 and αT was assessed by propidium iodide (PI) assay. Briefly, for PI assay, the cultures in 96 wells were stained with PI (7 μM) for one hour prior to the end of the incubation period (24 h. 7 days and 14 days). At the end of the incubation period, the fluorescence intensity was determined and expressed relative to cultures treated with 0.2% Triton X-100 (to permeabilize all cells). The fluorescence signal was measured by a fluorescence plate reader (Molecular Devices, USA) at 530-nm excitation ⁄ 645-nm emission to quantify cell membrane damage as described elsewhere [28].
Detection of Cell Survival
DS and euploid cortical neurons were pre-treated with varying concentrations of αT and γT3 (1-100 μM) for one hour at 37°C, followed by addition of H2O2 (100 μM) to the cells and a further incubation for 24 hours at 37°C before cell viability and apoptosis were assessed. Cell viability was assessed using MTT assay. Briefly, the cell culture media was loaded with 0.5 mg/mL MTT to detect any decrease in the cell metabolic activity using MTT reduction assay following standard procedures [29]. Meanwhile, the rate of apoptosis was measured using the ssDNA ELISA kit as described previously [20]. In addition, cell viability was also assessed utilizing the PI assay as described above. For cell imaging, DS neuron cultures were stained with of Neu-N (neuron- specific nuclear protein) antibody to confirm the results shown by the MTT and ssDNA ELISA assay. Briefly, cells were fixed with 4% paraformaldehyde before being permeabilized with 0.25% Triton in PBS for 30 mins. The cells were then washed with PBS, followed by incubation with goat serum at room temperature to block unspecific binding site, and incubation with mouse Neu-N antibody (Chemicon, USA) in 1:200 dilution overnight at 4°C. Subsequently, cultures were washed with PBS and incubated with anti-mouse Alexa Fluor 488 (Molecular Probes, The Netherlands) in 1:2000 dilution for 1 hr at room temperature. After washing with PBS, nuclei were counterstained with Hoescht before visualization under fluorescence microscope (Nikon, Japan) at 40× magnification. The intracellular production of ROS was determined using DCFH-DA assay. Non-fluorescent DCFH-DA was permeable to cell membrane and oxidation of hydroperoxides produced fluorescent 2',7'-dichlorofluorescein (DCF), which was detected by fluorescence plate reader at 485 nm excitation/530 nm emission [30].
Determination of Vitamin E Uptake by HPLC
The uptake of γT3 and αT was analyzed using reverse-phase high performance liquid chromatography (HPLC) Fluorescent EM 330 nm, EX 294 nm detector (Shimadzu, Japan) as described previously [20]. Concentration peaks of the samples were compared with tocotrienol rich fraction (TRF) standard and the concentrations of αT and γT3 uptake in cells were calculated as μM/106 cells.
SDS-PAGE and Western Blot
Western blot of DS and euploid (normal) cortical neurons in various treatment groups were used to elucidate the expression of proteins involved in the apoptosis signaling pathway including p53, Bax, Bcl-2, cPKC (for detection of common isoforms PKC-α, PKC-β and PKC-γ) and PKC-δ; while β-actin were used as housekeeping protein and loading control. A maximum protective dosage of 10 μM γT3 and αT was used to test if this concentration could induce apoptosis in DS and euploid neurons. The western blots were performed as previously described.
Statistical Analysis
Each experiment of cultures in microplates was carried out in triplicate wells with at least three independent cultures. The data were reported as mean ± SD of at least three experiments. Comparison between groups was made by the Student's t test, one-way ANOVA and Bonferroni adjustment as well as two-way ANOVA for multiple comparisons. p < 0.05 was considered as statistically significant for Student t-test whereas p < 0.0001 was considered as statistically significant for multi-factor comparisons.
Results and discussion
From the PI assays, 1 μM and 10 μM of γT3 and αT maintained cell viability but did not improve cell survival when it was added to the DS culture for 14 days, γT3 was cytotoxic to DS cortical neurons at concentration ≥ 100 μM compared to αT, with increased apoptosis of 25-35% and 5-8% respectively [Figure 1 (a)]. For neurons incubated with γT3, more apoptotic cells were observed at 50 μM, and at 100 μM almost all of the cells undergo apoptosis. From Figure 1 (b), a short 1 day incubation of γT3 at 10 μM and 100 μM in DS neurons was only able to reduced apoptosis by 10%. The previous landmark study has shown that αT was able to attenuate apoptosis and improve cell viability [11], whereas prolonged incubation time of γT3 up to 14 days increased membrane damage and apoptosis to DS neurons, as detected from the PI assay at a dose dependent manner. Similar to our previous studies, long term incubation of γT3 was shown to be cytotoxic to neurons at high dose (≥ 100 μM) while αT showed minor toxic effects to human neurons as illustrated in Figure 1 (a) and Figure 1 (b)[20,21]. Fluorescence detection of DCF showed that γT3 act as a free radical scavenger by reducing ROS generated by H2O2 in a dose dependent manner at concentration ≤ 1000 μM of γT3 after one hour H2O2 exposure [Figure 1 (c)]. The HPLC analysis revealed that the uptake of γT3 was higher compared to the uptake of αT in neurons [Figure 1 (d)]. The absorption of αT had been associated to ATP-binding cassette (ABC) transporter (MDR1) which acts as a cellular exporter of tocopherols to ApoA-I and HDL by ABCA1-dependent and ABCA1-independent processes [31,32]. Hence, this may explain why high doses of αT did not exhibit toxicity to neurons, as αT was effluxed from the cells by ABC transporter 1 (MDR1) to eliminate excess αT. On the other hand, γT3 was not selectively transported by TTP. Thus, excess γT3 was not being effluxed from the cells, leading to the accumulation of γT3 that might exacerbate cell death and contribute to the possible toxicity of γT3 at high dose.
Figure 1. (a) The effects of α-tocopherol (αT) and γ-tocotrienol (γT3) towards DS neuronal cell death for 14 days in vitro as detected by Propidium Iodide (PI) assay. *Denotes P < 0.05 compared to control while # denotes P < 0.05 when γT3 was compared to αT at their corresponding concentration. The data are presented as mean ± SD, n = 6. (b) The effects of γT3 at various incubation periods (1 day, 4 days and 14 days). *Denotes P < 0.05 compared to the control, while # denotes P < 0.05 compared to day 1 and $ denotes P < 0.05 compared to day 4. The data are presented as mean ± SD, n = 3. (c) The effects of γT3 on ROS generated by H2O2 in DS neurons with intracellular accumulation of hydroperoxides after one hour exposure to the indicated concentration of H2O2 measured by DCFH oxidation to DCF. *Denotes P < 0.05 compared to control while # denotes P < 0.05 compared to 10 μM γT3. The data are presented as mean ± SD, n = 3. (d) Uptake of α-tocopherol (αT) and γ-tocotrienol (γT3) in human cortical neurons measured by HPLC. The cellular uptake of γT3 and αT was significantly higher than the untreated control, with the uptake of γT3 significantly higher than that of αT. *Denotes P < 0.05 compared to the control, while # denotes P < 0.05 when γT3 uptake of γT3 incubated cultures were compared with αT uptake of αT incubated cultures. The data are presented as mean ± SD, n = 3.
In both DS and euploid neurons, the pre-treatment of γT3 and αT at concentration ≤ 50 μM was able to reduce cell death induced by H2O2 [Figure 2 (a) and Figure 3 (a)] and increase cell viability [Figure 2 (b) and Figure 3 (b)]. Figure 2 (c) and Figure 3 (c) shows that both pre-treatments of up to 10 μM γT3 and 50 μM αT were able to attenuate apoptosis in H2O2-induced DS and euploid neurons respectively. However, the effectiveness of γT3 and αT pre-treatment was more pronounced in euploid neurons compared to DS neurons, as two-way ANOVA analysis showed that vitamin E isomer type and concentration contributed significantly to cell viability and apoptosis rate in euploid neurons but not in DS neurons. This indicated that the protective effect of the vitamin E in euploid neurons is dose and isomer type dependent, while protective effects of vitamin E in DS neurons is not dependent on dose and isomer type. Figure 4 shows the morphology of H2O2-treated DS neurons as stained by Neu-N, a marker for differentiated neurons [33] which had undergone apoptosis; pre-treatment with 10 μM of αT or γT3 retained some of the neurons' viability but cell morphologies were not fully maintained. Reduced Neu-N expression in differentiated neurons indicated perturbed cell morphology induced by H2O2 assault, which pre-treatment of either αT or γT3 were not substantial enough to protect cells from oxidative assault. However, comparatively αT seems to be more protective to the DS neurons than γT3 as more cells were stained with Neu-N, although the staining was dim. This is not surprising since the previous study has shown that in vitro DS cortical neuron culture had higher sensitivity to H2O2-induced oxidative damage compared to euploid neurons [27]. H2O2 directly induces cellular damage and has been reported to induce parallel apoptosis and autophagy [34], making it more difficult for αT and γT3 to protect the cells especially if the pre-treatment incubation period was short. Since our previous studies have shown that high dose of vitamin E compounds the toxic effect of H2O2, here we further investigate whether non-lethal doses of high concentration αT and γT3 will further exacerbate the detrimental effects of H2O2 via the mitochondrial Bcl-2 family pathway or the PKC signalling pathway as both cPKC and PKC-δ are redox sensitive and were reported to be involved in the initiation of apoptosis signalling [35,36].
Figure 2. The effects of α-tocopherol (αT) and γ-tocotrienol (γT3) against H2O2-induced cell death in human DS neuron cultures, whereby (a) the cell death was determined using propidium iodide (PI) assay, (b) the cell viability was determined using MTT assay, (c) the apoptosis assay was determined using ELISA kits for ssDNA. The neurons were pre-treated with varying concentrations of αT and γT3 for one hour before the exposure to 100 μM H2O2 for 24 hours at 37°C. *Denotes P < 0.05 compared to control, † denotes P < 0.0001 compared to H2O2. The data are presented as mean ± SD, from 3 independent experiments of triplicate wells (n = 9). One-way ANOVA showed that there are significant differences between groups, F13, 83 = 14.47, P < 0.001. Meanwhile, two-way ANOVA showed both types of vitamin E isomer and vitamin E concentration are not significant factors contributing to the cell survival and no significant interaction between types of vitamin E isomer and vitamin E concentration (F5, 71 = 0.45).
Figure 3. The effects of α-tocopherol (αT) and γ-tocotrienol (γT3) against H2O2-induced cell loss in human euploid neuron cultures, whereby (a) the cell death was determined using propidium iodide (PI) assay, (b) the cell viability was determined using MTT assay, and (c) the apoptosis assay was determined using ELISA kits for ssDNA. The neurons were pre-treated with varying concentrations of αT and γT3 for one hour before the exposure to 100 μM H2O2 for 24 hours at 37°C. *Denotes P < 0.05 compared to control, † denotes P < 0.0001 compared to H2O2. The data are presented as mean ± SD, from 3 independent experiments of triplicate wells (n = 9). One-way ANOVA showed that there are significant differences between groups, F13, 83 = 9.81, P < 0.001. Two-way ANOVA showed both types of vitamin E isomer and vitamin E concentration are significant factors contributing to the cell survival and no significant interaction between types of vitamin E isomer and vitamin E concentration (F5, 71 = 27.15, P < 0.0001).
Figure 4. Fluorescence image of DS neurons stained with anti Neu-N antibody in various treatments: control, 100 μM of H2O2 only, 10 μM of αT or γT3 pre-treatment followed by H2O2 exposure
From our investigation, euploid neurons showed a completely different protein expression profile compared to DS neurons. In the control group, DS neurons were shown to have lower Bcl-2/Bax ratio and p53 expression compared to euploid neurons, while cPKC and PKC-δ expressions were higher in DS neurons. Figure 5 (a) (i), Figure 5 (a) (ii) and Figure 6a revealed that for DS neurons, pre-treatment of γT3 followed by H2O2 has significantly lower Bcl-2/Bax ratio than the controls, whereas other treatment showed changes which were not statistically significant. This result suggests that γT3 does not contribute to the survival of DS neurons under H2O2 assault via the Bcl-2/Bax heterodimer complex formation. However, the analysis of Western blot for the euploid neurons in Figure 5 (b) (i), Figure 5 (b) (ii) and Figure 6 (a) showed a different picture: Bcl-2/Bax ratio increased significantly in euploid neurons when neurons were pre-treated with either γT3 or αT followed by H2O2 which suggested that γT3 pre-treatment attenuated apoptosis and improved cell survival of normal euploid neurons. However, p53 expression was not significantly different across various treatments in both DS and euploid neurons, as depicted in Figure 5 (a) (iii), Figure 5 (b) (iii) and Figure 6 (b). Nevertheless, the comparison of p53 expression between DS and euploid neurons showed lower p53 expression in DS neurons for these treatment groups: control, H2O2, γT3 followed by H2O2 and γT3 treatments only. Taking it all together, these results show that lower Bcl-2/Bax ratio DS neurons in the untreated groups, was in agreement with the results from previous study which reported that fetal DS neurons had increased Bax and p53 expressions mediated by the transcription factor est-2 when treated with H2O2 [37]. On the other hand, another study reported that APO-1, caspase-3 and Bcl-2 protein expression levels were unaltered in the fetal DS neurons [38]. A previous study also stated that incubation of αT induced the up-regulation of Bcl-2 as preventive effects from neuronal cell death [39]. Thus, treatment of only αT and γT3 without the presence of H2O2 in human neuron did not show pro-apoptosis tendency (from the Bcl-2/Bax ratio and p53 expression) compared to rat cerebellar culture as reported previously [21].
Figure 5. Bcl-2, Bax, p53, cPKC and PKC-δ were differentially expressed in euploid and DS neurons which were given various treatments in the following fashion: untreated control, incubation of neurons with H2O2 for 24 hours (H2O2), incubation of neurons with γT3 (10 μM) for 24 hours (γT3), one hour of γT3 (10 μM) pre-treatment in neurons followed by H2O2 incubation for 24 hours (γT3 + H2O2), incubation of neurons with αT (10 μM) for 24 hours (αT) and one hour of αT (10 μM) pre-treatment in neurons, followed by H2O2 incubation for 24 hours (αT + H2O2). (a) Western blot of Bcl-2, Bax, p53, cPKC and PKC in DS neurons; (b) Western blot of the same proteins in euploid neurons.
Figure 6. Densitometric analysis of Bcl-2 and Bax as Bcl-2/Bax ratio in (a), p53 in (b), cPKC in (c) and PKC-δ in (iv). * Denotes to P < 0.05 compared to control, † denotes P < 0.05 compared to H2O2-treated neurons, ‡ denotes P < 0.05 compared to cells pre-incubated with γT3 or αT followed by H2O2 treatment while # denotes P < 0.05 compares to euploid neurons.
Aberrant expression of PKC signalling has been reported in fetal DS post-mortem tissues [40], while DS patients' fibroblast was reported to be hyposensitive to PKC [41]. However, both studies did not specify the location of PKC isoforms involvement. From previous study, cPKC was shown to be activated when exposed to oxidative stress in neuronal death induced by ischemia, hypoxia and exitotoxicity [42], whereas other studies revealed that an increase in PKC-δ expression was needed for glutamate-induced neuronal death [43], Parkinson's disease model [44] and AGE-induced neuronal death [45] as well as H2O2-induced oxidative stress [46]. Across the various treatment groups, the expression of cPKC was higher in DS neurons compared to euploid neurons. However, the cPKC expression of DS neurons was down-regulated in all other treatment groups (γT3 followed by H2O2, γT3, αT followed by H2O2, and αT), as shown in Figure 5 (a) (iv) and Figure 6 (c). From Figure 5 (a) (v) and Figure 6 (d), DS neurons showed a 2-fold increase of PKC-δ expression in H2O2-treated neurons, suggesting an accumulation of PKC-δ in the cytosol, which signified pro-apoptotic activities in neurons [44] were suppressed by the pre-treatment of γT3, while the pre-treatment of αT did not alter PKC-δ expression. In normal euploid neurons, H2O2 induced increased cPKC expression [Figure 5 (b) (iv) and Figure 6 (c)] but suppressed PKC-δ [Figure 5 (b) (v) and Figure 6 (d)]. Pre-treatment of γT3 suppressed the cPKC expression but elevated the PKC-δ expression; while the pre-treatment of αT was found to increase the cPKC expression but down-regulated PKC-δ expression [Figure 6 (c) and Figure 6 (d)]. αT has been known to inhibit PKC-α activities [47,48] which was not shown in euploid neurons pre-treated with αT in Figure 5 (b) (iv) and Figure 6 (c). Meanwhile a high concentration of αT (500 μM) has been shown to inhibit PKC-δ activation in AGE-induced neuronal death [45] in which lower dose of αT pretreatment in euploid neurons showed similar result [Figure 5 (b) (v) and Figure 6 (d)]. However, the incubation of only γT3 also showed a decrease in cPKC, similar to a previous study which showed that γT3 suppressed PKC-α expression [19]. This suggests that besides functioning as an antioxidant, γT3 might also play a role in modulating PKC-δ expression as PKC-δ is a redox sensitive molecule.
This study revealed that in DS neurons, even though γT3 pre-treatment provided initial slight improvement in neuron viability, the protection from both αT and γT3 pre-treatment was not substantial to protect DS neurons from H2O2 assault. Furthermore, pre-treatment of γT3 would reduce the Bcl-2/Bax ratio that indicates cell survival while αT pre-treatment did not suppress pro-apoptotic PKC-δ expression in the cells. However, in non-oxidative stress condition, αT and γT3 did not exert strong pro-apoptosis tendency in human DS and euploid neurons compared to our previous studies in rat neurons [21]. DS neurons has been shown to have chronic overexpression of S100B, in which oxidation of S100B preferentially induced the neurotrophic processes (which is beneficial for cell survival and differentiation) over neuroinflammation processes. However treatment with antioxidants such as vitamin E interrupts this feedback and leads to increase glial activation and cell death [25]. This may explain the reason why in DS neurons, αT and γT3 at the concentrations used in this study may aggravate cellular damages in a highly susceptible neuronal cell model subjected to oxidative stress. Our present results further underlie the importance of more study to be done on the safety of vitamin E supplementation in neurodegenerative diseases such as DS and AD. Although γT3 act as a free radical scavenger which could quench ROS generated from H2O2, it may also synergistically induce apoptosis and autophagy through the mitochondrial death pathway, including the Bcl-2 family proteins [49]. Meanwhile, our study also showed that αT has a different mechanism of action compared to γT3, which remains to be further elucidated.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
SMT carried out the most of the experiments, performed the statistical analysis and drafted the manuscript. WZWN and MM played a major role in the experimental procedures of this study and revised the manuscript. CS carried out the cell imaging work and help revised the manuscript. Meanwhile, GMT extracted the vitamin E isomers and tested the purity of the vitamin E isomers. All authors have read and approved the final manuscript.
We are grateful to Dr Santiago Barambio, Yolanda Trejo and Raquel Lopez from Tutor Medica Clinics, Barcelona, and Dr Nuria Toran from Hospital de la Vall d'Hebron, Barcelona, for their assistance in providing the tissue samples for this work. This study was funded by the Ministry of Science, Technology and Innovation Malaysia under the Intensified Research Prioritized Area (IRPA) grant 06-02-02-002/PR0008/09-07, SAF2009-13093-C02-02 from MICINN and RD06/0013/1004 from ISCIII, Spain.
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Int J Cancer 2010, 127:1823-1832. PubMed Abstract | Publisher Full Text
Rickmann M, Vaquero EC, Malagelada JR, Molero X: Tocotrienols induce apoptosis and autophagy in rat pancreatic stellate cells through the mitochondrial death pathway.
Gastroenterol 2007, 132:2518-2532. Publisher Full Text
Sylvester PW, McIntyre BS, Gapor A, Briski KP: Vitamin E inhibition of normal mammary epithelial cell growth is associated with a reduction in protein kinase Calpha activation.
Cell Prolif 2001, 34:347-357. PubMed Abstract | Publisher Full Text
Mazlan M, Sue Mian T, Mat Top G, Zurinah Wan NW: Comparative effects of alpha-tocopherol and gamma-tocotrienol against hydrogen peroxide induced apoptosis on primary-cultured astrocytes.
J Neurol Science 2006, 243:5-12. Publisher Full Text
Then SM, Mazlan M, Gapor MT, Wan Ngah WZ: Is vitamin E toxic to neurons?
Cell Mol Neurobiol 2009, 29:485-496. PubMed Abstract | Publisher Full Text
Shang F, Lu M, Dudek E, Reddan J, Taylor A: Vitamin C and vitamin E restore the resistance of GSH-depleted lens cells to H2O2.
Free Radic Biol Med 2003, 34:521-530. PubMed Abstract | Publisher Full Text
Zana M, Janka Z, Kalman J: Oxidative stress: a bridge between Down's syndrome and Alzheimer's disease.
Neurobiol Aging 2007, 28:648-676. PubMed Abstract | Publisher Full Text
Esposito G, Imitola J, Lu J, Filippis D, Scuderi C, Ganesh VS, Folkerth R, Hecht J, Shin SJ, Luvone T, Chesnut J, Steardo L, Sheen V: Genomic and functional profiling of human Down syndrome neural progenitors implicates S100B and aquaporin 4 in cell injury.
Bialowas-McGoey LA, Lesicka A, Whitaker-Azmitia PM: Vitamin E increases S100B-mediated microGLIA activation in an S100B-overexpressing mouse model of pathological aging.
Glia 2008, 56:1780-1790. PubMed Abstract | Publisher Full Text
Choo YM, Ma AN, Basiron Y: A method of chromotagraphic isolation for vitamin E isomers.
European Patent EP1122250; United States Patent 6656358
Sebastia J, Cristofol R, Pertusa M, Vilchez D, Toran N, Barambio S, Rodriguez-Farre E, Sanfeliu C: Down's syndrome astrocytes have greater antioxidant capacity than euploid astrocytes.
Eur J Neurosci 2004, 20:2355-2366. PubMed Abstract | Publisher Full Text
Rosa R, Sanfeliu C, Suñol C, Pomés A, Rodríguez-Farré E, Schousboe A, Frandsen A: The mechanism for hexachlorocyclohexane-induced cytotoxicity and changes in intracellular Ca2+ homeostasis in cultured cerebellar granule cell neurons is different for the c and d isomers.
Toxicol Appl Pharmacol 1997, 142:31-39. PubMed Abstract | Publisher Full Text
Hansen MB, Nielsen SE, Berg K: Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill.
J Immunol Methods 1989, 119:203-210. PubMed Abstract | Publisher Full Text
Sebastia J, Cristofol R, Martin M, Rodriguez-Farre E, Sanfeliu C: Evaluation of fluorescent dyes for measuring intracellular glutathione content in primary cultures of human neurons and neuroblastoma SH-SY5Y.
Reboul E, Trompier D, Moussa M, Klein A, Landrier J, Chimini G, Borel P: ATP-binding cassette transporter A1 is significantly involved in the intestinal absorption of alpha- and gamma-tocopherol but not in that of retinyl palmitate in mice.
Am J Clin Nutr 2009, 89:177-184. PubMed Abstract | Publisher Full Text
Oram J, Vaugham A, Stocker R: ATP-binding cassette transporter A1 mediates cellular secretion of alpha-tocopherol.
J Biol Chem 2001, 276:39898-39902. PubMed Abstract | Publisher Full Text
Mullen RJ, Buck CR, Smith AM: NeuN, a neuronal specific nuclear protein in vertabrates.
Development 1992, 116:201-211. PubMed Abstract | Publisher Full Text
Zhang H, Kong X, Kang J, Su J, Zhong J, Sun L: Oxidative stress induces parallel autophagy and mitochondria dysfunction in human glioma U251 cells.
Toxicol Sci 2009, 110:376-388. PubMed Abstract | Publisher Full Text
Majumder P, Mishra N, Sun X, Bharti A, Kharbanda S, Saxena S, Kufe D: Targeting of protein kinase C delta to mitochondria in the oxidative stress response.
Cell Growth Differ 2001, 12:465-470. PubMed Abstract | Publisher Full Text
Yamaguchi T, Miki Y, Yoshida K: Protein kinase C δ activates IκB-kinase α to induce the p53 tumor suppressor in response to oxidative stress.
Cell Signalling 2007, 19:2088-2097. PubMed Abstract | Publisher Full Text
Helguera P, Pelsman A, Pigino G, Wolvetang E, Head E, Busciglio J: est-2 promotes the activation of a mitochondrial death pathway in Down's syndrome neurons.
Engidawork E, Balic N, Juranville JF, Fountoulakis M, Dierssen M, G L: Unaltered expression of Fas (CD95/APO-1), caspase-3, Bcl-2 and annexins in brains of fetal Down syndrome: evidence against increased apoptosis.
J Neural Transm Suppl 2001, 61:149-162. PubMed Abstract
Numakawa Y, Numakawa T, Matsumoto T, Yagasaki Y, Kumamaru E, Kunugi H, Taguchi T, Niki E: Vitamin E protected cultured cortical neurons from oxidative stress-induced cell death through activation of mitogen-activated protein kinase and phosphatidylinositol 3-kinase.
J Neurochem 2006, 97:1191-1202. PubMed Abstract | Publisher Full Text
Peyrl A, Weitzdoerfer R, Gulesserian T, Fountoulakis M, Lubec G: Aberrant expression of signaling-related proteins 14-3-3 gamma and RACK1 in fetal Down syndrome brain (trisomy 21).
Electrophoresis 2002, 23:152-157. PubMed Abstract | Publisher Full Text
Govoni S, Bergamaschi S, Gasparini L, Quaglia C, Racchi M, Cattaneo E, Binetti G, Bianchetti A, Giovetti F, Battaini F, Trabuechi M: Fibroblasts of patients affected by Down's syndrome oversecrete amyloid precursor protein and are hyporesponsive to protein kinase C stimulation.
Neurology 1996, 47:1069-1075. PubMed Abstract
Durkin JP, Tremblay R, Chakravarthy B, Mealing G, Morley P, Small D, Song D: Evidence that the early loss of membrane protein kinase C is a necessary step in the excitatory amino acid-induced death of primary cortical neurons.
J Neurochem 1997, 68:1400-1412. PubMed Abstract | Publisher Full Text
Choi B, Hur E, Lee J, Jun D, Kim K: Protein kinase C delta-mediated proteasomal degradation of MAP kinase phosphatase-1 contributes to glutamate-induced neuronal cell death.
J Cell Sci 2006, 119:1329-1340. PubMed Abstract | Publisher Full Text
Kanthasamy AG, Anantharam V, Zhang D, Latchoumycandane C, Jin H, Kaul S, Kanthasamy A: A novel peptide inhibitor targeted to caspase-3 cleavage site of a proapoptotic kinase protein kinase C delta (PKCδ) protects against dopaminergic neuronal degeneration in Parkinson's disease models.
Free Radic Biol & Med 2006, 41:1578-1589. PubMed Abstract | Publisher Full Text
Nitti M, d'Abramo C, Traverso N, Verzola D, Garibotto G, Poggi A, Odetti P, Cottalasso D, Marinari UM, Maria A, Pronzato MA, Domenicotti C: Central role of PKC-delta in glycoxidation-dependent apoptosis of human neurons.
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Carvour M, Song C, Kaul S, Anantharam V, Kanthasamy A, Kanthasamy A: Chronic low-dose oxidative stress induces caspase-3-dependent PKCdelta proteolytic activation and apoptosis in a cell culture model of dopaminergic neurodegeneration.
Ann N Y Acad Sci 2008, 1139:197-205. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Ricciarelli R, Tasinato A, Clement S, Ozer NK, Boscoboinik D, Azzi A: alpha-Tocopherol specifically inactivates cellular protein kinase C a by changing its phosphorylation state.
Biochem J 1998, 334:243-249. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Carter CA, Kane CJ: Therapeutic potential of natural compounds that regulate the activity of protein kinase C.
Curr Med Chem 2004, 11:2883-2902. PubMed Abstract | Publisher Full Text
Yap WN, Chang PN, Han HY, Lee DTW, Ling MT, Wong YC, Yap YL: gamma-tocotrienol suppresses prostate cancer cell proliferation and invasion through multiple signalling pathways.
Brit J Cancer 2008, 99:1832-1841. PubMed Abstract | Publisher Full Text | PubMed Central Full Text


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Gary Shaw calls new ProElite "a joke" & discusses Kimbo Slice's boxing debut
"I think it's laughable. People who are running it are jokes. It's not real and the only game in town is the UFC." – Gary Shaw
Fight Nerd correspondents Syl Peterkin and Elizabeth Amadeo caught up with Gary Shaw at the NYC press conference for an upcoming boxing pay-per-view entitled "Believe It or Not: Hopkins vs. Dawson". Shaw, one of the key people behind the original ProElite and EliteXC, discussed his thoughts on the light-heavyweight world championship match between Bernard Hopkins and "Bad" Chad Dawson, and gave us his opinion on the new version of ProElite which he does not look at fondly. We also ask if he was asked to be involved with the new company and if he thinks anyone can ever overcome the UFC. Lastly, we ask Shaw's thoughts on Kimbo Slice's boxing debut and if he still has what it takes to be a star in combat sports.

subset: commoncrawl


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Monster Hunter Freedom 3 Finally Falls From 1st Place
January 20, 2011 - How do you make a PSP Monster Hunter game fall from the top spot in Japan's weekly sales chart? With another PSP game! And it doesn't hurt if you have some widespread hardware shortages too.
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The operations are set to continue in the sequel to the hit PSP title, Metal Gear Solid: Portable Ops.


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A man who stabbed an officer in the head last year outside the police station pleaded guilty Wednesday while telling the judge medication is controlling his bipolar disorder.
John Heffner, 42, entered a guilty plea to one count of felonious assault on a police officer, a first degree felony. He faces up to 11 years in prison when he is sentenced, which is scheduled for Feb. 20.
Heffner also told the judge the medication does not cloud his mind, and he was entering the plea fully aware of what he was doing.
Heffner previously pleaded not guilty by reason of insanity and had sought at least two psychological exams, but the results apparently didn't come back in his favor. The judge ruled he should stand trial.
The charge was for the March 27 stabbing of Sgt. Dave Gillispie outside the front door to the Lima Police Department on Market Street. Heffner called officers outside and charged once they stepped out. He used a screwdriver to stab Gillispie in the head.
Gillispie managed to draw his weapon despite the injury and fired several shots, striking Heffner twice, to stop the attack. Gillispie was taken to the hospital following the attack. The sergeant did not have a life-threatening injury.
Heffner also made a statement to a detective about the incident. Police officials said Heffner wanted to act in such a way that a policeman would shoot and kill him, a phenomenon known as "suicide by cop."
By Greg Sowinski - The Lima News, Ohio (MCT)
©2014 The Lima News (Lima, Ohio)
Visit The Lima News (Lima, Ohio) at www.limaohio.com
Distributed by MCT Information Services


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Jean-Louis Borloo
Head of Union of Democrats and Independents party Jean-Louis Borloo... NachrichtenfotoAm Telefon,Drahtlose Technologie,Frankreich,Ganzkörperansicht,Handy,Jean-Louis Borloo ,Kommunikation,Nationalversammlung,Paris,Politik,Politische Partei,Reden,Telefon,VertikalPhotographer AFPCollection: AFP 2012 AFPHead of Union of Democrats and Independents (UDI) party Jean-Louis Borloo speaks on the phone on November 21, 2012 in Paris at the National Assembly. Former campaign staff members of defeated candidate for the leadership of the UMP right-wing opposition party, Francois Fillon, announced that they are contesting the election results. AFP PHOTO / THOMAS SAMSON (Photo credit should read THOMAS SAMSON/AFP/Getty Images)


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Sunset Beach Resort and Waterpark
Montego Bay, Jamaica, Caribbean
Sunset Beach Resort and Waterpark Introduction
Amidst swaying palms, shimmering water and beautiful beaches, the all-inclusive Sunset Beach Resort and Waterpark offers something for everyone. Sparkling restaurants and bars with sleek, contemporary designs await guests with an array of new, all-inclusive dining options. Playful and exciting, you'll splash your way through the magnificent, Pirates Paradise waterpark- with twist, turns, rides and slides for all ages. Simmer in the beachfront hot tub, or stroll through the tropical gardens. Spend a day pampering yourself with a treatment in the Spa or partake in one of many land and water sports. Friendly and pleasant staff will ensure that you will have a holiday to remember.
Sunset Beach Resort and Waterpark Facilities
• Resort View room for up to 3 adults consists of • Kingsize or two double beds • Rollaway bed (on request) • Bath, shower and wc • Hairdryer • Desk • Iron and iron board • Tea and coffee making facilities • Satellite TV • Safety deposit box • Air conditioning • Balcony • Bay View room and Ocean View rooms available at a supplement
• Tennis • Basketball • Volleyball • Minigolf • Shuffleboard • Giant chess and checkers • Pirates' Paradise Waterpark including giant waterslides and lagoon • Watersports including sailboats, paddle boats, kayaking and snorkelling • Giant Jacuzzi • Spa with a range of treatments
All Inclusive means
• Meals: Breakfast, lunch and dinner with buffet and full board options. • Drinks: Unlimited selected beverages and cocktails from the six bars. • Activities: Tennis (Floodlit). Air conditioned gym which includes aerobics and weights. Non-motorised watersports. • Entertainment: Daily activities. Nightly entertainment including Caribbean floor shows, fashion shows and beach party nights. Splash and play at the Pirates' Paradise Water Park. • For Children: Kids' Club (2-12 years) running an activities program. Pirates Cove Teen Centre (13-19 years) with entertainment program.
Extra Value
• Free non-motorised watersports
• 3 swimming pools, one is Adults only with swim up bar • Gift shops • WiFi access available
For Children
• Kids' Club (2-12 years) with extensive activities • Teens Club (13-19 years) including billiards, ping pong, poster competitions, dance contests and Disco night
Key Facts
• 430 rooms • Lift • Child age: 2-16 years • Some facilities and activities may incur a local charge
• On private peninsula
Meals and Drinks
• Banana Walk buffet restaurant including healthy options and kids' station • La Bella Vita Italian restaurant (dress code/reservations apply) • Silk Road serving a contemporary Asian menu • Byron's Caribbean fusion • Sunset Beach Grill serving jerk chicken, burgers, hotdogs etc until late • Sundowner Bar also with swim up bar • Lobby Bar • Tamarind bar • Ortanique Bar • 2 Beach bars
Selected Highlights
• Families • Spa Treatments • Watersports • Central location
If you are interested in this holiday, please select your preferred travel options from the panel on the left.


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F1 in Austin
- F-1 Is Over But Traffic Woes Remain
- UT Football And F-1 Take Over Austin
- Traffic Hassles Pile Up As F1 Crowds Head Downtown
- Public Safety A Priority For F1
- City of Austin, Cap Metro, ABIA Prepare for Formula 1
- F1 Fans Pushing Austin Flood Victims Out Of Hotels
- Austin Fan Fest Line-up Announced
- Multiple Big Events To Snarl Downtown Traffic This Weekend
- UT's 2013 Football Schedule Shows OSU Game During F1 Weekend Race
- Austin Airport Busy With F1 Attendee Departures
- Austin F1 Race Over, but Debate on Tax Subsidies Continues
- F1 Weekend is a Tax Windfall -- For Some
- Central Texas Hotels Already Gearing Up For 2013 Formula One Race
- Retailers Outside Austin See Spike In Sales For F1
- 117,000 Fans Attend Austin's Inaugural Formula 1 US Grand Prix
- Brit Lewis Hamilton Wins Formula 1 U.S. Grand Prix In Austin
- Things Run Smoothly At F1 Track
- 82,000 Attend Formula 1 U.S. Grand Prix Qualifying In Austin
- Business Down For Some Downtown Austin Stores Due To Fan Fest
- Fans Mellow So Far At Austin Fan Fest
- F1 Fan Fest Draws Big Crowds To Downtown Austin
- Austinites Check Out 1st Day Of F1
- Download F1 Road Closures Map
- Turning Laps In The Sky For Austin F1
- KEYE TV News Tests the F1 Shuttles For You
- 24 Hours Emergency Operations Center Open For F1 Weekend
- F1 Austin Fan Fest Begins Today
- Officials Make Final Austin F1 Traffic Plans
- Fan Fest Chaos Poses Challenges For Downtown Austin Businesses
- Austin Fan Fest Street Closures Continue
- Law Enforcement Offers Austin F1 Risk Assessment
- Formula 1 Track Shows Off Race Day Emergency Plan
- Cap Metro Makes F1 Backup Plans As Union Threatens Strike
- South Austin Neighborhoods Not Happy With F1 Helipads
- Austin Council Eyes F1 Helipad Code Changes After Neighborhood Outcry
- Austin Council Annexes F1 Race Track Land
- Bike Route Planned From Downtown Austin To Formula 1 Track
- Austin Formula 1 Racetrack Opens Saturday For Running Event
- Capital Metro To Provide Free Bus Service Around Downtown For F1 Weekend
- F1 Marks First Lap, Leaders to Discuss Preparedness
- Helipads Approved In South Austin For F1
- F1 Announces More Bands For Race Weekend
- Police, Nonprofits Bracing For Human Trafficking Increase
- How To Avoid Buying Counterfeit Tickets
- Austin F1 Transportation Plan Announced
- Austin Race Track Gets Green Light As Deadline Nears
- Pflugerville Wants To Study Impact Of F1
- Nelly Joins Aerosmith As Top Acts At F1 Grand Prix Concerts
- Condo Owners Wanting To Make Some Quick Cash Could Be Fined Thousands
- Festivals Planned During Austin's F1 Race Weekend
- Tens Of Thousands To Flock To Downtown Austin For F1 Grand Prix Events
- F1 Bringing Aerosmith To Austin During Race Weekend
- Texas Gov. Perry Promotes Austin F1 Race During Italy Trip
- Workers Needed For F1 Race
Austin Council Annexes F1 Race Track Land
Updated: Thursday, November 8 2012, 06:17 PM CST
The Austin City Council approved the annexation of the racetrack that will hold the upcoming Formula One race.
It's about 1,500 acres of land east of Austin-Bergstrom International Airport. Most of the land is owned by Circuit of the Americas, the operator of the track.
Austin's first Formula One race is scheduled Nov. 16-18.
Austin Fan Fest
View Austin Fan Fest in a larger map
Austin Grand Prix Parking
View Formula 1 Austin Grand Prix: Parking in a larger map


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By ajcolores
Posted in IndustryMusicVideo | Tags : Arizona, gadsen, gadsen records, indie, music, Phoenix, Record Label
In episode four of #WeezyWednesdays we catch up with Tune, Mack and Tyga at the studio. After that, we follow the YMCMB crew to Label where Wayne recently hosted a party.
Previously: Lil Wayne – The Moment


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Gaw, a blogger at The Dabbler, can't stomach traditional Cockney cuisine:
Jellied eels somehow seem a more appropriate accompaniment to a funeral than baked meats. I can see the mourners exchanging regretful comments between lugubrious spoonfuls of silvery eel and golden jelly, turning away occasionally to suck on the odd piece of protuberant cartilage.
… I can't think of another instance where I haven't liked the local, traditional, everyday food of a place (unless it's featured tripe): wurstl, pizza, croque monsieur, balti, burgers, gallettes, chips and mayo, waffles, herring (pickled and smoked), tapas, salt beef, stovies, noodles, fish and chips, tacos, kebabs, knishes, dosas, phô, pasties, pelmeni (OK, perhaps I'll make an exception for that last one – the Russian equivalent of the Cockney pie). And so on. No, I really would like to enjoy Cockney grub. But I just can't – it's simply no good.
Just stay a while for the spotted dick and bubble and squeak. And yes, I grew up on both, which may explain this. George Orwell's classic defense is here. It reads like a blog-post avant-la-lettre.
(Photo of jellied eels by Flickr user Malias)


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25th Annviersary Menu
Patrick Hession had a stunning intuition the first time he heard Maynard Ferguson play. Even at that early age, he intuitively knew he would play the lead book on Maynards band - and even more, that he would be Maynards last lead player. That powerful psychic glimpse of the future helped propel Patrick into a career most lead trumpet players have only experienced in their dreams.
For the last six years of Maynard Fergusons career up until his very last gig Patrick Hession played the lead trumpet book on Maynards band. Thats the longest time anyone ever exclusively played the lead book without a break on the band. And as legendary Maynard lead trumpet player Stan Mark has said publicly many times, Patrick was the perfect choice to play the book in those most challenging of times. Stan, a vereran of 10 years (a record!) in Maynards trumpet section, knows that of which he speaks. Patrick had to play not only the lead book, but also cover Maynards solos from time to time - including playing the entire re-tooled MF HIT MEDLEY every night.
When Patrick first visited the shop years ago, Dave told him he played with his head tilted up too far, and that his throat was closed off enough that Monette mouthpieces would not work for him. The next time Dave saw the band, Patrick had switched to a bent mouthpiece, which allowed him to lower his head and open his throat. Dave was delighted, and they started a friendship and working relationship that has grown more and more rewarding every year since!
In this interview, Patrick tells funny stories from the road, gives us some insight into the crazy schedule the band endured (and also enjoyed!), and shares his thoughts on how to survive playing extreme lead trumpet. Patrick wowed the audience at our 25th anniversary party with several displays of his outrageous power, range and musicianship. Videos including Patrick in performance are found elsewhere in this newsletter and on our YouTube site. We hope you enjoy Patricks video interview. Thanks for all the great hangs, Patrick!
6918 NE 79th Court Portland, Oregon 97218 USA (503)255-5552 FAX (503)255-5593
Copyright ©2006 David G. Monette Corporation


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Whether it's been called the Radiation Laboratory (Rad Lab), Lawrence Berkeley Laboratory (LBL), Lawrence Berkeley National Laboratory (LBNL), or simply Berkeley Lab, this facility has been on the cutting edge of science and technology research for over 80 years.
Located on a 200-acre site in the hills above the University of California's Berkeley campus, LBNL is the oldest U.S. Department of Energy (DOE) national laboratory.
Managed by the University of California (UC) for the DOE, the Laboratory conducts unclassified research that fosters groundbreaking fundamental science that enables transformational solutions for energy and environment challenges, using interdisciplinary teams and by creating advanced new tools for scientific discovery.


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Hi everyone,so I am trying to mod this pair of silnylon "hipsacks" which I carry behind the hipbelt pockets on either side of my SMD Comet for more hip pocket storage space. They are currently kept closed by cord that just loops through the top (like a normal stuff sack) and is then fastened with a cord lock. Here they are:However, it's not easy to tighten the cordlock one-handed and I have lost some gloves that way (sack not properly tightened)... I have replaced the loop with some stretch cord and ideally I would like to be able to tighten it down with one hand. The best option that I can think of are 1-handed cord locks, you know, the type that is itself attached to the little stuff sack.Like this one here (I think):So can you think of what the most minimalist type of cord lock is that would do the job? Where could I get it? Would you recommend 1/16" Shock Cord like zpacks is selling it to keep that sack shut? How should I fix that cord lock in place?Your help is very much appreciated. As you can probably tell, I am a noob in MYOG matters.
Hi JohannesAll sorts of bits and pieces of hardware and fabrics etc can be easily bought from OWFINC, Seattlefabrics, and others. They will ship overseas too. Google those names.Cheers
Hi Roger,thank you for the suggestion. I had checked out OWFINC's website (and the others) before posting and was hoping for some advice from the experience of people who have already played around with that sort of thing in real life. Maybe my question it too trivial.peace,johannes
Here is exactly what you are looking for.FECWS - with slot on each side for tie down with flat draw string Black, Desert Tan, Foliage .34 .22http://owfinc.com/Hardware/phardware.asp#CordlocksSome thin webbing or drawcord to hold it to the seam and some 1mm drawcord or 1/16"shockcord. The zpacks stuff should work but will stretch a lot so almost all of the cord will be hanging off the cordlock. Should mean that it seals very well, but could have limitations i hadn't considered.-Tim
You can easily use any cordlocks this way, attaching them with a loop that goes through the same hole as the drawcord. This is the way it has been done before all the special cordlocks appeared.
thank you! I´ve rigged something up that looks like it might work.
Yep, to echo Jan, securing the cord lock by the same cinch hole is the way to go. A lot of mil/tactical gear use the ITW Cordloc you pictured in this fashion, and in my experience it works better than using the auxiliary slots.
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October 16, 2002
In May 2001, 7.8 million persons worked at multiple jobs in the United States, a figure representing 5.7 percent of all workers. Why did these persons choose to work more than one job?
Results from a supplement to the May 2001 Current Population Survey reveal that more than 1 in 3 moonlighters worked multiple jobs in order to earn extra money, a category that could include saving for the future or getting extra money to buy something special. An additional 27.8 percent moonlighted in order to meet current expenses or pay off debt.
Among the other common reasons for working multiple jobs, enjoyment of the second job was reported by 17.4 percent, and 4.6 percent wanted to build a business or get experience in a different job.
These data are from a supplement to the Current Population Survey. For additional information on reasons for moonlighting, read "Twenty-first century moonlighters," Issues in Labor Statistics (PDF 169K), Summary 02-07.
Bureau of Labor Statistics, U.S. Department of Labor, The Economics Daily, Moonlighting in 2001 on the Internet at http://www.bls.gov/opub/ted/2002/oct/wk2/art02.htm (visited October 10, 2015).
New estimates of personal taxes in Consumer Expenditure Survey
In 2013, the Consumer Expenditure Survey improved its personal tax data.
Trends in long-term unemployment
Long-term unemployment reached historically high levels following the recession of 2007–2009.
Housing: before, during, and after the Great Recession
looks at consumer expenditures on household items, employment in residential construction, prices for household items, and injuries in occupations involved in building and maintaining our homes.


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June 16, 2007 11:41 AM EDT
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Glad to be "green"
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My wife and I started an all-natural (green) cleaning business. Although I admit we`re kinda cheaters...We aren`t 100% green. As of now we`re mainly targeting commercial customers and they don`t seem to really go for the green cleaning products as much as say upper middle-to-upper income households would. So we`re trying to stay as green as possible, unless they request a commercial toxin based product, and/or we just simply cannot find a natural product to do the particular job.
May 26, 2007 12:46 PM EDT
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May 24, 2007 8:32 AM EDT
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May 22, 2007 11:42 AM EDT
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Not exactly. Having a sign does not make your vehicle 100% deductible for tax purposes. You need to keep a very detailed mileage log for that.You can get a background check online, or through your local police department. Even people in the military have records - no joke. So that would be some type of document I would print out, make copies of, and offer to customers along w/the list of references, resume, etc.???
May 22, 2007 11:21 AM EDT
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Help with Marketing for my new business
in the forum
I work for tip income and I absolutely would NOT include tip information in your pricing. That is a completely rude turnoff. I would, however, post on your vehicle about your service. As you drive around town you get free advertising.Your type of business does not need a website at this time. When you have an income and want to sell your green products, then you need a website. Your money can be better spent otherwise.Are you bonded and insured? I`ve wanted to hire some occasional help and changed my mind when I found out they were not bonded. Also, do you provide a scrubbing service? So many cleaning companies damp mop the floors and simply are pushing the dirt around.Do you require a top to bottom cleaning first with maintenance cleaning afterwards? Will you do one room cleaning only if that`s all the customer wants?Have you had a police background check so that you can offer that assurance to your customers?Jillybeans The magnet sign on the vehicle was something we were really thinking about. Besides great advertisement, wouldn`t that also enable us to write the vehicle off on our taxes as a business expense? Bonded and insured we are working on that. Actually, that was probably going to be another post...the best ways to go about this, suggestions for companies to go through, etc. Scrubbing service, I`m not sure...Can you give more detail on what exactly that would be? Do you mean like wiping down baseboards, getting up grime, mildew, etc., and not just mopping over? That`s what we were planning, do an initial cleaning, likely charge more that first time or two, then after it`s been given a through, treat all normal, repeat cleanings after that as maintenance cleanings. For police background...I`m not sure how to go about that? I just got out of the military 2 months ago, had a secret clearance when I was in Iraq a few years ago...So are you saying this is just something you tell the customers? Or would you have an actual document from the police dpt. showing a background check? If you mean having an actual hardcopy of something, then no we would have to do that, if that`s not what you mean, then I believe we should be ok there. Thanks for all the suggestions!
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Microsoft will soon debut the new Windows 8.1 OS update and company CEO Steve Ballmer hasn't missed the occasion to praise it during the Worldwide Partner Conference 2013 in Houston.
Ballmer emphasizes that Windows 8.1 is proof that Microsoft is trying to get much closer to its users, so the return of the Start button plus the rest of improvements included in the first Windows 8 overhaul are nothing more than a living proof that the company is now listening to consumers.
"We're going to spend a lot of time today also talking about Windows 8.1, because we listened and we learned, and as good as Windows 8 is I would just encourage you to really take a look at the Windows 8.1," Ballmer said in the beginning of keynote.
"It really speaks to the feedback that we've gotten. It builds on the exciting new user interface, the embrace of new platforms, the new programming model. But we've also suggested the feedback that we've gotten from people on how it works in the enterprise, what we need for serious desktop users, and I think you'll be incredibly impressed by the work on Windows 8.1."
As far as Windows 8 is concerned, Microsoft's big honcho claimed that everything goes according to the plan, despite all statistics pointing to a rather slow update for this particular operating system.
Ballmer pointed out that all Microsoft executives are very satisfied with Windows 8, so everybody expects more people to make the move to this version in the coming months, especially thanks to the arrival of the brand new Windows 8.1 update.
"Windows 8 was nothing short of the most remarkable replatforming of Windows basically since 1995. And we've gotten a lot of feedback on Windows 8. I think we did a heck of a good job on Windows 8," he said.


text:
well i think its cause the ford already has the problem circled
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An organization that has touted itself as being all about saving women's lives made it clear that women's lives don't mean much in the face of political opposition to abortion.
The 800 pound gorilla of breast cancer marketing and research, the Susan G. Komen Foundation, announced it was severing ties with Planned Parenthood because of a new policy that bars grants to organizations that are under investigation by local, state or federal authorities. According to Komen, because Rep. Cliff Sterns (R-FL) launched a congressional investigation into whether or not Planned Parenthood was using public money on abortions, that means no more grant money. Period.
The Republican attacks on Planned Parenthood have been relentless and baseless. They are nothing short of vindictive, politically-motivated smears targeting an organization that makes it possible for women to have access to safe, affordable health care. They are fueled by an animus toward women and a belief that women's bodies are, essentially, community property at best.
The Komen Foundation grants to Planned Parenthood totaled about $680,000 last year and $580,000 the year before, with that money mainly going to breast-cancer screening and other breast-health services. Translated to actual women's lives, it looks like this: almost 170,000 clinical breast exams and more than 6,400 mammograms.
It's a move that is not only disappointing, it is deadly. "We are alarmed and saddened that the Susan G. Komen for the Cure Foundation appears to have succumbed to political pressure. Our greatest desire is for Komen to reconsider this policy and recommit to the partnership on which so many women count," said Cecile Richards, president of Planned Parenthood Federation of America.
Planned Parenthood and its affiliates are looking beyond political pressure and focused only on how to help save women's lives. "While this is deeply disturbing and disappointing, we want to assure women who rely on Planned Parenthood for breast care that we're still here for them, and we always will be. The new fund we're launching to support these services will ensure that the Komen Foundation's decision doesn't jeopardize women's health," added Richards.
Thanks to a grant by Amy and Lee Fikes' Foundation Planned Parenthood established a Breast Health Emergency Fund to provide an immediate source of funding to ensure that Planned Parenthood health centers can continue to provide critical cancer screenings and care that actually saves lives.
The Susan G. Komen Foundation will slap a pink ribbon on just about anything, from NFL football jerseys to a bucket of Kentucky Fried Chicken. Yet they won't stand up to anti-abortion zealots. The result puts real women's lives at risk. If you care about women, and if you want to support an organization that knows that providing women access to life-saving cancer screens means being stronger than political pressure from the religious right, I suggest sending that support directly to Planned Parenthood. The Susan G. Komen Foundation, it seems, has other concerns.
Related Stories:
BREAKING: Komen Cancer Charity Pulls Aid To Planned Parenthood
Read more: abortion, breast cancer, planned parenthood, Susan G. Komen, war on women, war_on_women
Photo from jdmith1021 via flickr.
I could deliver a sermon on what's wrong with PP, but everyone deserves to have that choice to make themselves.
I realize that Komen G for the cure and Nancy Brinker has really messed things up, but this non profit organization located in the United States of America is right neck and neck with her with her antics and pink washing schemes, only she hides behind her MD title.
Petition signed!!! I used to walk in our support for Susan G. Komen Foundation, but not any more. I will Never donate or walk for them again. From now on my donation will go directly to Planned Parenthood.
Thanks for sharing :-(
It's not too uncommon for something that starts out small with good intentions to balloon into something that greedy people use for their own self-enrichment more than anything. When a so-called charity gets this big and SELLS stuff, that sounds like a business to me ... a for-profit business. When the heads of these "charities" make astronomical wages and benefits, that's when the whole thing starts to smell bad. When you get "political" people from high places working in charities or religious organizations, you have trouble ... and the money usually gets steered towards their favorite "earmarked" projects, etc.
Sometimes things need to come around full circle. When the rich get involved, the rich get richer and the disadvantaged get screwed over.
SGK always smelled of a sham to me.
Shame on Komen Foundation... And I will never wear a Pink Ribbon again... Back stabbers and hypocrites is what they are, afraid to defend the women of this country and go up against the rich, uncaring politicians who are making us women their victims...
As women, we are the birthers of yesterday, today & tomorrow. And as such, we alone have the ONLY RIGHT to dictate what goes on with our bodies, except maybe the Creator. This stopping of funding is a hypocracy when there are so much governmental funds just bring put into the raising, including paying foster parents, of children of drug users, unfit parents, least to mention just plain orphans. Yes, too much is going there & more will follow if we have our womanly rights taken away. Ladies, it is time to band together, like those of another era who won us our right to vote. There is funding folks, it is just the government is spending elswhere & the hell with us in this country, or how much did they waste on a senseless war the killed our future fathers and mothers needlessly... Yes, needlessly because it was more than the Twin Towers, or Bin Laden, it was, in reality, a war to secure oile rights, nothing more.
Komen donations aren't up in my area. the only thing up is the number of women banding together and boycotting anything with a pink ribbon and pulling out in droves from the race for the cure. Sad when political nonsense is more important than a mother who might be able to see her children grow up had she had access to a mammogram. I would have died had i not had access to a pap smear. Grow up people and quit holding the American public hostage as a tool for dirty politics.
Supported in their decision, seems the Susan Goodman Komen Foundation's donations are up over 100% since their withdrawal from PP.
Disclaimer: The views expressed above are solely those of the author and may
not reflect those of
Care2, Inc., its employees or advertisers.
This is very nice. I love this kind of people who help animals in need.
Here is the BBC I-Player link, I'm not sure if it works in the USA though (I hope it does). http…
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text:
Minutes 06.04.09
University of Virginia School of Medicine
Curriculum Committee
Minutes 06.04.09
Pediatric Conference Room, 4:00 p.m.
Present (underlined) were: Gretchen Arnold, Dan Becker, Robert Bloodgood, Megan Bray, Eugene Corbett, Thomas Gampper, Wendy Golden, Donald Innes (Chair), Keith Littlewood, Veronica Michaelsen, Mohan Nadkarni, Chris Peterson, Jerry Short, Linda Waggoner-Fountain, Bill Wilson, Mary Kate Worden, Lisa Herrmann, Evan Lapinsky, Debra Reed (secretary) Guests: Brad Bradenham, Pam Herrington
- Announcements: The Roanoke Times reports that the new medical school - Virginia Tech Carilion School of Medicine, in Roanoke has received preliminary accreditation and can now begin to recruit students. The school will open by August 2010 with approximately 42 students.
Psychiatric Medicine Clerkship Review. Pam Herrington met with the Curriculum Committee to discuss the recently submitted Psychiatric Medicine clerkship report. Dr. Herrington outlined the program and how it works at the various sites (Roanoke Carilion, Salem VA, Western State and at UVA). Students are require to complete the on line patient logs and a Passport. The Curriculum Committee encourages continued review of the patient logs and passports by the course director at midpoint of the rotation to determine student compliance and to alert students by e-mail if at least 50% has not been completed. All of the sites provide an inpatient experience. Electives in an outpatient psychiatric setting are being discussed but are difficult to arrange. There is one two week selective in an outpatient setting at the present time but it is not optimal due to the short time frame students can follow a patient.
The clerkship has approximately 14 students per month - equally spread between Roanoke/Salem (2-3 each), UVA (4-5) and Western State (4-5).
ED-2 items are listed on the passport and must be signed off prior to the student taking the shelf exam. A completed passport is often referred to as the "ticket" to take the shelf exam.
The course director is aware of variation in the orientation packets/lecture notes at the various sites. A consolidated outline to be used at all sites would be ideal but will take some time to prepare.
The Clerkship Director communicates with faculty both inside and outside of UVA via e-mail. One on one face-to-face communication is more common within the University of Virginia at faculty meetings and on the floors.
The Clerkship Coordinator position is occupied by Rebecca Johnson who is an outstanding addition to the course.
The Curriculum Committee thanks Dr. Herrington for her detailed report and the well run Psychiatric Medicine clerkship. The Committee also offers support to the clerkship if the director feels it is needed.
Institutional Programs and Educational Activities to Address the Needs of Gay, Lesbian, Bisexual and Transgender (GLBT) Students and Patients. Brad Bradenham brought a recommendation proposal from the Joint AAMC-GSA and AAMC-OSR regarding Institutional Programs and Educational Activities to address the needs of GLBT Students and Patients. Meredith Rogers (SMD 2010) attended a meeting of the GSA where this topic was addressed. She has since met with SMEC (Brad Bradenham) to develop a specific plan to implement curriculum revisions to address these issues. A plan, including modifications in SIM, PoM1 and 2 CPD, the 2nd year Psychiatric Medicine course, the Sexual Health Clinical Connection as well as the possible addition of an away elective at the Cllen-Lorde Community Health Center were discussed. Development of a faculty development component also seems appropriate. It was noted by the committee (Bill Wilson) that these topics are covered in the current curriculum but are perhaps not as apparent as they could be. The Committee supports Meredith Rogers in her attempt to integrate this topic into the curriculum and encourages her to work with course directors, Dana Redick (Clinical Connections) and particularly Fern Hauck (Cultural Competency) to further enhance this topic.
"Next Generation" Preliminary Calendar. Proposed drafts of possible "Next Generation" calendars were distributed. The number of weeks for each component of the curriculum are still not fixed and will be changed as necessary. The number of weeks assigned now is in line with those at other medical schools who have a Systems Based Curriculum.
The possible addition of a "Bugs and Drugs" course was briefly discussed. This course would not replace these items in the individual systems but rather be an introductory component/overview prior to the systems.
The Committee was asked to consider the topics, orders, etc. and share comments with Don Innes.
Donald Innes


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text:
Weekend may bring snow to Upstate
ANDERSON - More snow may be on the way for Anderson County.
State and local officials are gearing up for an approaching storm that forecasters say could dump four to six inches of snow across northeast Georgia and Upstate South Carolina on Monday.
"We are getting all of our equipment ready to go," said Dusty Turner, a state Department of Transportation assistant district maintenance engineer.
Turner said hundreds of transportation employers will be deployed throughout the Upstate on Sunday to place salt and sand on Interstate 85 and other major highways before conditions deteriorate.
AnMed Health officials also are keeping a close eye on the weather, spokeswoman Heidi Charalambous said Friday.
"Anytime there's a chance of inclement weather, AnMed Health works closely with Anderson County Emergency Services to monitor the storm and take the proper precautions," Charalambous said.
"Engineering crews are prepared to come in Sunday and pre-treat the grounds with ice melt. They'll also start blocking any areas that could become slip hazards, including steps and steep inclines. The goal is to get ahead of the bad weather as much as possible and prevent accidents before they happen."
AnMed Health has a number of physicians' practices across the Upstate and Charalambous said patients with appointments will be contacted if the weather causes closings or delays.
Oconee Physician Practices also will notify news media if the weather leads to any changes in business hours.
"We want to remind our patients to look to these local news resources for information and prevent them going out on the roads when they are potentially unsafe," said Matthew Hedtke, executive director of Oconee Physician Practices.
The National Weather Service forecast calls for sunny skies and breezy conditions Saturday with a high of 43 in Anderson. Sunday also will be sunny with the temperature topping out at 39 degrees
Snow should start falling before dawn Monday and continue until midday, when there is a chance that the precipitation will change over to sleet.
The extended outlook calls for cold temperatures next week with highs in the 30s and overnight lows dipping into the teens.
Subscribe to the Independent Mail
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Monday-Friday: 6am-4:30pm / Saturday: 7am-11am / Sunday: 7am-2pm


text:
The Administration of Barack Obama initially tried to downplay reports that the Internal Revenue Service placed undue scrutiny on conservative groups over the past several years, but a damning Treasury report on the IRS-Tea Party scandal released this week introduces Administration foot to Administration mouth.
The official narrative from top Federal officials that the IRS abuse of conservatives was relegated to the agency's Cincinnati-based Determinations Unit was destroyed by the report's repeated references to the IRS's Washington, D.C.-based Exempt Organizations Unit and guidance specialists also working for the IRS in Washington. In fact, the Treasury Inspector General's report notes that 10 of 12 IRS offices involved in the dubious practices are in Washington.
The White House was forced to issue a fickle statement on the explosive report.
"The report's findings are intolerable and inexcusable," Obama said in a statement. "The federal government must conduct itself in a way that's worthy of the public's trust, and that's especially true for the IRS. The IRS must apply the law in a fair and impartial way, and its employees must act with utmost integrity. This report shows that some of its employees failed that test."
ABC, which recently took the Obama Administration to task over the Benghazi, Libya, cover-up, has been at the forefront of coverage on much of the IRS scandal as well. Most recently, ABC political analyst Trey Hardin said that as more information comes to light about the IRS ordeal, it will likely become apparent that top Administration officials were involved in green lighting the agency's underhanded practices.
"I will tell you this on the IRS front. I've worked in this town for over 20 years in the White House and on Capitol Hill, and I can say with a very strong sense of certainty that there are people very close to this president that not only knew what the IRS were doing but authorized it," he said, tiptoeing around directly blaming Obama for the scandal.
"It simply just does not happen at an agency level like that without political advisers likely in the West Wing certainly connected to the president's ongoing campaign organization," he added.


text:
Leicestershire's retiring wicketkeeper Paul Nixon has urged England's selectors to pick his team-mate James Taylor again soon.
Taylor made his England debut in last Thursday's one-day victory over Ireland in Dublin but has been overlooked for the forthcoming five-match NatWest series against India.
Nixon, who will retire after next month's Champions League in India following Leicestershire's gripping Friends Life t20 final win over Somerset, believes that Taylor is a more likely match-winner than Warwickshire's Jonathan Trott.
"James Taylor should have been playing for England weeks ago," Nixon said. "I rate Jonathan Trott highly as a high-class Test batsman, as good as anyone in the game at the moment and his record proves that.
"But for one-day cricket I don't think Jonathan's game suits to bat through and try to go hard at the end. I think Jonathan Trott will do a job for England on a very juicy wicket where a workmanlike innings is needed. Ian Bell bats three for Warwickshire, Jonathan Trott bats four.
"James Taylor averages 50 in one-day cricket and 34-35 in Twenty20 cricket. He's a finisher, he hits the ball out of the ground. Anyone who was here a few weeks ago against Warwickshire saw how he got an amazing 50 and won the game.
"He cleared the boundary with ease and that's what he can do. He's got every option. He's got Dilshan scoops, he can hit you down the ground straight, he walks across his stumps and hits you out of the ground.
"This guy has got magic, I promise you. In his understanding of where he is in his game, he's a 34-year-old experienced pro. He knows his options, he knows his game.
"He ticks every box. If you say right the ball is swinging away at 84mph and this is the field where do you score your runs, he knows. He's a little professor of his industry."


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Our giveaway today is perfect for the upcoming holiday season!!!
Party Inspiration, Printables + Sweet Party Supplies
Kim Stoegbauer, Creative Director + Founder of The TomKat Studio Blog + Shop. Kim frequently styles for HGTV and DIY Network, and brand partners including Pottery Barn Kids, Barbie, Shutterfly + Cricut. Her work has been featured by Martha Stewart, Hostess with the Mostess and Project Nursery, as well as in national magazines including Parenting, Babytalk, Celebrate, Mingle + Fresh Style.
Jenni says
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I just became a fan of TomKat om facebook! =)
travelgirl18 says
I'm a FB fan of TomKat!!
travelgirl18 says
It's so hard to pick a favorite outfit from shabby apple but I've narrowed it down to two, LOL! Bette D and Tender is the night. Thank you Fawnice
Michelle says
I'm a fan of TomKat on facebook
Michelle says
I am now a fan of Shabby Apple on facebook
Michelle says
It is so hard to pick a favorite but I I love the Champs Elysees and the Ingrid
Devon and Josh says
I am LOVIN" the Red Fox dress!
Devon and Josh says
I am & have been a fan of TomKat! I love you gals!
Devon and Josh says
Just became a fan of Shabby Apple! LOVE their dresses!
Debbie Thompson says
I love the color and style of Alto Sax.
Debbie Thompson says
I'm also a exiting fan and have been for some time of TomKat Studio….
Stephanie Ann says
I'm your fan on Facebook!
Stephanie Ann says
I like Shabby Apple on facebook!
Stephanie Ann says
I love the black wrap dress…shoot I already forgot what it's called! But i love love love it!
Tonia V says
If I had to choose just one, it would be the Miles dress. I'm a sucker for all things houndstooth… Thanks TomKat for introducing me to Shabby Apple!
Destiny says
Fan of TomKate
Destiny says
I must have the VP in BLACK!! WOW:0)
jill says
I am already a fan of Tomkat on FB
jill says
I really like the ballerina dress.
Mary Craig says
I love the Sierra Nevada dress!
Mary Craig says
I'm a fan of TomKat on FB!
mary Craig says
I'm a fan of Shabby Apple on FB!
Amy says
i'm a fan of shabby apple on facebook
Girl from Pennsylvania says
I absolutely LOVE the last dress you posted a picture of. Burnt orange is one of my favorite colors perhaps bc I went to school at the University of Tennessee!
Crystal says
Ribbon Falls is my current fave.
Crystal says
Shabby Apple Facebook fan
Crystal says
TomKat Facebook fan
coolmom:) says
I am already a huge fan! But really love finding out about Shabby Apple!! Telling ALL my friends! Excited for your contest as well!
Shannon says
I'm a fan on FB – thanks for the chance to win!
Shannon says
And now I'm a fan of Shabby Apple. I am totally in love with the Queen of Hearts girls dress – beautiful!
Monica P. says
I love the orange cider dress and am dreaming of the Cleopatra.
Monica P. says
I am a FB Fan of TomKat!
Veronica says
There are too many choices! I'd have to say Sierra Nevada, Tanglewood, L'Artiste or Le Soleil.
vbbradford at gmail dot com
Veronica says
I'm a fan of TomKat on Facebook
vbbradford at gmail dot com
Veronica says
I'm a fan of Shabby Apple
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Dawn says
I really love Black Oak
Dawn says
I am a fan of Shabby Apple on FB!
Dawn says
I am already a fan of TomKat on FB!
mymomistoocoolforasu says
I love the little black dress named "New Orleans" which happens to be one of my favorite places in the world!
mymomistoocoolforasu says
I "like" TomKat on Facebook
mymomistoocoolforasu says
I "like" Shabby Apple on Facebook!
jeavin blog says
A BIG THANK YOU! I am so excited to get a new dress from Shabby Apple! I just emailed Toni. You guys are the best!


text:
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text:
Little Moreton Hall
How to get here
By train
Kidsgrove 3 miles; Congleton 4½ miles
By road
on east side of A34. From M6 exit 17 follow signs for Congleton and join A34 southbound (signed Newcastle) from Congleton
Parking: 100 yards
By foot
leave the Macclesfield canal at bridge 86. Map to Hall displayed on towpath
By bus
services from Alsager to Congleton (passing close to Kidsgrove), infrequent (not Sundays)
By cycle
List view
Map view
- Key
- National Trust
place to visit
- Location
- Cluster of pins Shows number of grouped pins. Zoom in to see individual pins


text:
If you think that you have seen more than you share fair of netbook by now, wait until you lay your eyes on this low power (in more ways than one) Datawind creation, the UbiSurfer9 being one of those ARM powered smartbooks that first started to pop up in early 2009.
Powered by an ARM processor this device comes as pretty low end portable computing solution, Datawind adding an extra two inches to the screen compared to their previously released UbiSurfer7 while retained pretty much the same hardware configuration that relies on an ARM processor as well as an unimpressive 128MB of RAM and 1GB of storage (expandable via a SD slot).
In case that you wonder how does the UbiSurfer9 manages to go by with such low system specs, the answer lies in the operating system used, Datawind's offering coming with Windows CE pre-installed.
However, the most important thing to note about the UbiSurfer9 is its connectivity options since this is actually built for Internet surfing, apart from WiFi this also coming in with an embedded GPRS wireless modem and a special data plan that brings 30 hours of surfing each month free of charge for the first year you own the product thanks to a Vodafone UK SIM.
After the first year passes by, the same plan is available for £29.99 per year, an unlimited traffic subscription being available for an additional £5.99 a month.
"We continue to see strong demand for low cost, connected netbooks. While many new products and form factors move towards higher costs, the need continues to grow for low cost devices that deliver web access at affordable rates," said Suneet S. Tuli, CEO of Datawind
"Studies show that one in four adults in the UK have never used the Internet.
We strive to help lower the cost of Internet access, while making the process even simpler," concluded the company's CEO.
As you probably guesses by now, this device will be available in the UK for now, the Datawind UbiSurfer9 coming to the Ideal TV home shopping channel this weekend, priced at £149.99. (via pocket-lint)