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Growing experimental and clinical data highlight a complex interaction among lipids, immune cells, and the hepatic inflammatory responses [1–4]. Accumulation of lipid leads to inflammatory cell infiltration and activation which promotes secondary tissue injury and organ dysfunction [1]. Key aspects in the regulation of this process remain unclear, particularly the intersection of lipid metabolism and immune cell function whether it be direct or indirect through hepatocellular stress/damage. Peroxisome proliferator activated receptor alpha (PPARα) is a nuclear hormone receptor associated with proliferation of peroxisomes in the hepatocytes of rodents in response to a number of naturally occurring as well as synthetic compounds [5]. PPARα is also a regulator of the production of a number of enzymes including acyl Coenzyme A oxidase (AOX) involved in the metabolism of fatty acids within the liver [6, 7]. As a result, mice deficient in this AOX present with an age dependent increase in the accumulation of hepatocellular fat or steatosis.
PPARα also plays a prominent role in inflammatory response [8–10]. For example, foam cell formation is reduced by the ligand-specific activation of PPARα in a model of hypercholesterolemia induced atherosclerosis [11]. Human monocyte-derived macrophages have also shown sensitivity to PPARα ligand activation with increased levels of apoptosis [9, 10, 12]. Further investigation has revealed an inhibitory effect of PPARα on the pro-inflammatory transcription factor nuclear factor kappa B (NFκB), a possible mechanism for its anti-inflammatory actions [13]. Jones et al. also report the presence of PPARα in CD4+ T lymphocytes in rodents [14]. As with macrophages, PPARα in T lymphocytes appears to regulate the activity of NFκB suggesting a common mechanism and role in immune cell function [14]. Interestingly, studies have also demonstrated a dysregulation of cytokine production in T lymphocytes from PPARα-deficient (PPARα−/−) mice whereby deficient cells produce significantly larger quantities of interferon gamma (IFNγ) in response to anti-CD3/anti-CD28 activation [15]. Such data would suggest that PPARα is capable of modulating the function and immunological response of a variety of immune cells from macrophages to T cells and therefore may play a significant role in the determination of T cell responsiveness in vivo.
Concanavalin A (ConA) is a plant lectin capable of inducing severe T cell mediated hepatitis in the mouse [16]. ConA activates CD1d-dependent intrahepatic natural killer T (NKT) cells to produce a number of proinflammatory mediators including tumor necrosis factor alpha, interleukin 4 (IL4), and IFNγ [17–19]. Given the presence of PPARα in T cells, its apparent regulation of T cell and macrophage activation, and its influence on hepatocellular lipid metabolism, PPARα lies at the unique nexus of lipid metabolism and immunological function. The current study was thus aimed at understanding the impact of PPARα in the complex setting of T cell-mediated hepatitis. To this end, we have administered ConA to wild type and PPARα−/− mice and revealed a surprising and profoundly protective effect of PPARα deficiency on ConA-mediated, T cell-dependent liver injury, a protection likely related to reductions in hepatic NKT cell number and function.
Eight to twelve week old male C57Bl/6 mice, PPARαdeficient (PPARα−/−) mice [20], or severe combined immunodeficient (SCID) mice on a C57Bl/6 background were purchased from Jackson Laboratories (Bar Harbor, ME). All animals were housed in specific pathogen free conditions with 12 h light/dark cycles and free access to food and water. All subsequent procedures described were approved by the Institutional Animal Care and Use Committee at the University of North Carolina at Chapel Hill and complied with the "Guide for the Care and Use of Laboratory Animals".
Lipopolysaccharide treatment
Male mice, either wild type or PPARα−/−, were administered lipopolysaccharide (LPS; 1 mg/kg, Sigma, St. Louis, MO) by intraperitoneal injection in 200 μl of normal saline or saline alone as control 6 h prior to sacrifice.
Concanavalin A (ConA) mediated hepatitis
Male mice, either wild type or PPARα−/−, were administered Concanavalin (ConA; Sigma, St. Louis, MO) at a dose of 15 mg/kg in sterile saline via tail vein injection as has previously been described [21]. Mice were then anesthetized with ketamine and xylazine (100 and 10 mg/kg respectively) 10 or 24 h following injection, the diaphragm severed to effect euthanasia, and serum and tissue collected.
α-Galactosylceramide (αGal) treatment
Male mice, either wild type or PPARα−/−, were administered αGal (Funakoshi, Tokyo, Japan) by intravenous injection at a dose of 10 μg/mouse through the tail vein as previously reported [2]. Mice were then euthanized 12 h as described above to assess liver injury and cytokine production.
Liver enzyme assessment
Blood was collected from the inferior vena cava from anesthetized mice 10 h following ConA administration into sterile microcentrifuge tubes. Blood was allowed to clot on ice for a period of 10 min after which it was centrifuged at 12,000×g allowing for collection of serum. Serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured by the Clinical Chemistry Laboratory at the University of North Carolina at Chapel Hill using standard techniques.
Histopathology and immunohistochemistry
Liver tissue was collected at the time of sacrifice and placed in 10% buffered formalin (Thermo-Fisher Scientific, Waltham, MA) at 4 °C for 24 h. After fixation, the tissue was embedded in paraffin and 7 μm thick sections cut. Sections were then deparaffinized, rehydrated, and stained with hematoxylin and eosin. Additionally, some sections were stained for the T cell marker, CD3ε (Thermo-Fisher Scientific), as previously described [22]. Sections were examined under routine light microscopy at 100× and 400× magnification and images captured using an Olympus DP70 digital camera.
Terminal UTP nick end labeling (TUNEL) staining
To assess liver cell death, deparaffinized sections were stained for DNA fragmentation using a commercially available kit (In situ cell death detection kit, Roche, Indianapolis, IN, Cat# 11684795910) according to the manufacturer's recommendations as previously described [21]. Stained sections were viewed by fluorescent microscopy and images capture with an Olympus DP70 digital camera. Five random high powered fields were observed and positive cells counted.
Hepatic triglyceride quantification
Liver triglycerides were quantified using kit from Sigma (Triglyceride Reagent, Cat.# T2449, St. Louis MO) according to the manufacturer's recommendations as previously described by our group [2]. Triglyceride content was normalized to wet weight of tissue used in the assay.
Real time polymerase chain reaction
Total RNA (5 μg) isolated with Trizol reagent (Thermo-Fisher) was reverse transcribed using a kit obtained from Applied Biosystems (High Capacity Reverse Transcription Kit Cat.# 4368814, Foster City, CA). For quantification of message expression, 250 ng of cDNA was amplified in a Eppendorf RealPlex2 using the primers listed in Table 1 (except IL15 where primers were purchased from Real Time Primers, Elkins Park, PA) in the presence of Sybr Green I (Maxima Sybr Green Reagent, Cat.# K0221, Applied Biosystems) using 45 cycles of a three step protocol, 95 °C for 10 s, 57 °C for 15 s, and 72 °C for 20 s. All message expression was normalized to the housekeeping gene β actin and expressed as gene expression relative to the wild type 0 h animals using the comparative ct method. Amplification of a single product was verified by analysis of post-amplification product dissociation temperatures (i.e. melt curves).
|Primer sequence
|For 5′-TGCCCGAACTACAGTCACGAAC-3′
Rev 5′-AGTGACCTCGCCTGGTGAAATG-3′
|Tumor necrosis factor alpha (TNFα)
|For 5′-AGCCCACGTAGCAAACCACCAA-3′
Rev 5′-ACACCCATTCCCTTCACAGAGCAAT-3′
|Interferon γ (IFNγ)
|For 5′-TCAAGTGGCATAGATGTGGAAGAA-3′
Rev 5′-TGGCTCTGCAGGATTTTCATG-3′
|Interleukin 12 p40 (IL12p40)
|For 5′-GGAAGCACGGCAGCAGAATA-3′
Rev 5′-AACTTGAGGGAGAAGTAGGAATGG-3′
|Interleukin 4 (IL4)
|For 5′-ACAGGAGAAGGGACGCCAT-3′
Rev 5′-GAAGCCCTACAGACGAGCTCA-3′
|Interleukin 5 (IL5)
|For 5′-AGCACAGTGGTGAAAGAGACCTT-3′
Rev 5′-TCCAATGCATAGCTGGTGATTT-3′
|Interleukin 10 (IL10)
|For 5′-GGTTGCCAAGCCTTATCGGA-3′
Rev 5′-ACCTGCTCCACTGCCTTGCT-3′
|Acyl-CoA oxidase (AOX)
|For 5′-CTTGTTCGCGCAAGTGAGG-3′
Rev 5′-CAGGATCCGACTGTTTACC-3′
|Cluster of differentiation 1d (CD1d)
|For 5′-TCCTAGAGGCAGGGAAGTCA-3′
Rev 5′-AGCATTTGGCAGGAAATCAC-3′
|Peroxisome proliferator activated receptor alpha (PPARα)
|For 5′-GTGGCTGCTATAATTTGCTGTG-3′
Rev 5′-GAAGGTGTCATCTGGATGGGT-3′′
|Liver fatty acid binding protein (LFABP)
|For 5′-GTGGTCCGCAATGAGTTCAC-3′
Rev 5′-GTATTGGTGATTGTGTCTCC-3′
|β actin
|For 5′-AGGTGTGCACCTTTTATTGGTCTCAA-3′
Rev 5′-TGTAGTAAGGTTTGGTCTCCCT-3′
Flow cytometry
Liver mononuclear cells and total splenocytes were obtained as described previously [2, 21]. Isolated cells were stained for the immune cell markers T cell receptor beta (TCRβ; BD Pharmingen, San Jose, CA), CD4 (Thermo-Fisher), pan natural killer cell (DX5; Thermo-Fisher), and the activation marker, CD69 (Thermo-Fisher) at a 1:100 dilution for 30 min at room temperature. For spleen cells, whole spleens were homogenized between glass slides centrifuged at 500×g, and filtered through a 30 μm sterile filter followed by staining with the above listed antibodies. Again, cells were stained with the above listed antibodies. Cells were then analyzed and relative numbers expressed by % of total mononuclear cells and/or % of total liver TCRβ+ cells in liver mononuclear cell fraction.
Enzyme-linked immunosorbent assay
Serum and/or tissue culture media IL12, IFNγ, or IL4 protein was determined using a kit from R&D systems (IL12, Cat#M1270; IFNγ, Cat#MIF00; and IL4, Cat#M4000B) per manufacturer's instruction as previously described [21]. Samples were compared to a standard curve and values expressed per mg of liver protein.
In vitro ConA activation
Wild type or PPARα−/− mononuclear cells were isolated as described previously. For activation studies, 1 × 105 liver mononuclear cells or spleen cells were incubated in a 96 well plate in 300 μl of RPMI media (Invitrogen) in the presence or absence of 1 μg/ml ConA (Sigma) for 72 h at 37 °C and 5% CO2. Following incubation, media was collected and assessed for IFNγ and IL4 protein by ELISA as described above.
SCID lymphocyte reconstitution
Total splenocytes (2 × 107) were isolated as described above from wild type and PPARα−/− mice. Red blood cells were removed by incubation in red blood cell lysis solution for 10 min at room temperature. Cell viability and number were assessed by trypan blue exclusion. Splenocytes (2 × 107) were resuspended in 100 μl of PBS and injected intravenously into SCID recipients through the tail vein. SCID mice administered PBS alone served as controls for these experiments. Seven days following reconstitution, animals were administered ConA (15 mg/kg). Ten hours later, serum and tissue were collected to assess T cell reconstitution, liver injury, and cytokine expression.
Characterization of PPARα−/− mice
PPARα is a known regulator of lipid metabolism with significant relevance to liver function [6, 23]. Figure 1 characterizes the impact of a loss of PPARα on hepatic lipid accumulation as well as lipid metabolizing/transporting gene expression. Ten week old PPARα deficient mice have increased microvesicular lipid accumulation as assessed by routine histopathology (Fig. 1a) and a significant increase in triglyceride content (Fig. 1b). Expression analysis confirms absence of PPARα in our knockout mice (Fig. 1c) and this loss correlates with reduced hepatic acyl-CoA oxidase (Fig. 1d) and liver fatty acid binding protein (Fig. 1e) expression as has previously been reported [6]. Together, these data are consistent with previous reports and highlight the impact of a loss of PPARα on the hepatic microenvironment and provide a platform to study its impact on ConA-induced, T cell-mediated tissue injury.
Deficiency in PPARα inhibits Concanavalin A (ConA)-mediated hepatitis
ConA administration is an established model of T cell-mediated hepatitis in rodents [16–19, 24]. Doses from 10 to 20 mg/kg body weight are associated with significant NKT cell-dependent hepatocellular injury [16, 21]. To determine the role that PPARα plays in ConA-mediated, T cell dependent liver injury, 10 week old wild type and PPARα−/− mice were given 15 mg/kg ConA by intravenous injection. Ten hours following this dose of ConA, serum ALT and AST levels were significantly elevated in wild type mice (Fig. 2a, b) with levels remaining elevated through 24 h post-injection. This increase in serum levels of ALT or AST was not observed in PPARα−/− mice 10 h post-injection (Fig. 2a, b). Consistent with serum measurements of liver injury, histopathological assessment of livers from ConA pre-treated wild type mice revealed large areas of necrosis with the appearance of inflammatory cell infiltrate (Fig. 2c). Examination of liver sections from PPARα−/− mice treated with ConA confirmed the protective effect of this deficiency.
ConA has also been shown to induce liver injury through the induction of hepatocellular apoptosis via a Fas-dependent mechanism [25–27]. To determine if the ConA-induced apoptotic cell death was also disrupted in PPARα−/− mice, liver sections from wild type and PPARα−/− mice were subjected to the TUNEL assay to assess DNA fragmentation, a marker of apoptotic cell death. Consistent with serum enzyme measures and histopathological signs of liver damage, wild type mice given ConA had increased numbers of TUNEL positive cell number when compared to their untreated controls at 10 and 24 h post-injection (Fig. 2d). In contrast, PPARα−/− livers were resistant to ConA-induced increases in hepatocellular apoptosis, a finding consistent with an absence of liver injury. Taken together, these data suggest that PPARα may be involved in the early development of ConA-induced, T cell mediated, liver injury in mice.
Splenic and hepatic T cells are activated in wild type and PPARα−/− mice in response to ConA
ConA is known to activate both peripheral and intrahepatic T cells [18, 19]. More specifically, the activation of intrahepatic CD4+ natural killer T (NKT) cells is a key component of ConA-induced liver injury [17]. Splenic and intrahepatic T cells from wild type and PPARα−/− mice were therefore isolated and stained for the T cell marker CD4 in combination with the early activation marker, CD69 [28]. As shown in Fig. 3, wild type and PPARα−/− splenic and intrahepatic CD4+ T cells were activated to similar levels 10 h following ConA administration. These data confirm that T cells from wild type and PPARα−/− mice respond similarly to ConA exposure.
Deficiency in PPARα alters ConA-induced cytokine expression in liver
A number of studies have demonstrated the ability of ConA to induce pro-inflammatory cytokine expression in the liver and the importance of this cytokine production for the development of hepatocellular injury [18, 19, 24, 29, 30]. Indeed, deletion of interleukin 4 (IL4) or interferon gamma (IFNγ) has been associated with substantial reductions in ConA-induced liver injury [18, 19]. Given the importance of cytokines to the development of ConA-induced hepatitis, it was hypothesized that the cytokine response would be impaired in PPARα−/− mice when compared to their wild type controls. As shown in Fig. 4, wild type mice given ConA present with significant increases in a number of inflammatory mediators associated with acute hepatitis including tumor necrosis factor alpha (TNFγ; Fig. 4a), certain Th1 type cytokines including interferon gamma (IFNγ; Fig. 4b), and interleukin 12 (IL12; Fig. 4c) and certain Th2 type cytokines including interleukin 4 (IL4; Fig. 4d) interleukin 5 (IL5; Fig. 4e), and interleukin 10 (IL10; Fig. 4f). PPARα−/− mice administered the same dose of ConA had reduced expression of key Th1 type cytokines involved in ConA mediated liver injury, specifically IFNγ but similar levels of Th2 type cytokines such as IL4 and IL5 when compared to their ConA-treated wild type controls. These data suggest that PPARα is involved, either directly or indirectly, in the activation of the Th1-dependent, IFNγ mediated, inflammatory response caused by ConA administration.
It is becoming increasingly apparent that certain transcription factors play important roles in the differentiation of T cells towards Th1 or Th2 phenotypes [31–33]. T-bet, a T box transcription factor primarily expressed in T cells, is associated with the expression of Th1-type cytokines including IFNγ [34]. Furthermore, activation of T-bet has been shown to be crucial to the development of ConA-mediated hepatitis [35]. Given the reduction in expression of IFNγ in PPARα−/− mice following ConA when compared to ConA-treated wild type controls, we tested the hypothesis that PPARα positively regulates expression of this Th1-associated transcription factor. T-bet expression is strongly up-regulated in the livers of wild type mice 10 h following ConA administration (Fig. 4g). In contrast, deficiency in PPARα prevents the up-regulation of this Th1-associated transcription factor in the liver (Fig. 4g). Together, these data, in conjunction with the reductions in cytokine expression, suggest that PPARα does indeed play a role, either directly or indirectly, in the activation of the Th1-associated transcription factor T-bet following ConA administration.
To better understand the defects that are associated with a deficiency in PPARα, wild type and PPARαdeficient liver and spleen mononuclear cells (MNCs) were isolated from untreated animals and cultured in the presence or absence of ConA (1 μg/ml) for 72 h. Media was then analyzed for the presence of IFNγ and IL4 by ELISA. As shown in Fig. 5, splenocytes and hepatic MNCs responded to ConA stimulation with the production of large quantities of both IFNγ and IL4. Absence of PPARα led to a significant reduction in IFNγ production by hepatic MNCs. Interestingly, production of IL4 by these hepatic MNCs was not affected by absence of PPARα. Moreover, splenic MNCs from PPARα deficient mice showed significant increases in both IFNγ and IL4 production when compared to similarly treated wild type MNCs. Together, these in vitro data further confirm the selective impairment of liver derived mononuclear cell production of IFNγ.
PPARα−/− mice have reduced numbers of liver NKT cells
ConA-mediated liver injury requires CD1d-dependent NKT cell activation [17]. Given the profound protection against ConA-induced liver injury associated with a deficiency in PPARα, we examined the NKT cell populations in untreated wild type and PPARα−/− mice. As shown in Fig. 6a, untreated wild type mice have significant numbers of TCRβ and pan-NK positive cells, NKT cells (4.8% of total liver mononuclear cells, 21.5% of hepatic TCRβ+ lymphocytes) in the liver. In contrast, PPARα−/− mice have significantly reduced numbers of NKT cells (1.13% of liver mononuclear cells, 7.9% of hepatic TCRβ+ lymphocytes) within the liver despite having similar levels of TCRβ positive and pan-NK negative cells (T cells) and TCRβ negative and pan-NK positive cells (NK cells). Taken together, these data implicate PPARα in the development, recruitment, or differentiation of hepatic NKT cells. Furthermore, these data provide a mechanism by which PPARα may regulate ConA-induced T cell-mediated hepatitis.
To further evaluate the functionality of NKT cells directly, wild type mice or PPARα−/− mice were administered alpha galactosylceramide (αGal), a potent and specific activator of NKT cells [36]. Twelve hours following injection, mice were sacrificed and serum and tissue collected for liver enzyme release and pro-inflammatory cytokine production. As shown in Fig. 6b, absence of PPARα resulted in reduced αGal-induced liver injury as assessed by serum ALT levels as well as a reduction in IFNγ gene expression (Fig. 6c) following αGal administration when compared to similarly treated wild type mice. These data further confirm the dysfunction of hepatic NKT cells within PPARα−/− mice.
PPARα−/− splenocytes are capable of restoring ConA-dependent liver injury in SCID mice
To determine if the reductions in NKT cell number was indeed due to the absence of PPARα within other cell populations and not to an absence of this transcription factor in NKT cells directly, SCID mice, which express normal levels of PPARα within the liver parenchymal and non-parenchymal cells, were reconstituted with total wild type or PPARα−/− splenocytes. Seven days following lymphocyte reconstitution, wild type mice, SCID mice, and SCID mice reconstituted with either wild type or PPARα−/− splenocytes were administered ConA (15 mg/kg). Reconstitution was verified by immunohistochemical detection of CD3ε in the spleen and liver. As shown in Fig. 7a, SCID mice have no CD3ε positive cells within the spleen or liver whereas SCID mice reconstituted with wild type or PPARα−/− splenocytes showed splenic and hepatic repopulation of CD3ε positive cells to a comparable to that of untreated wild type mice. Ten hours following ConA administration, wild type mice showed significant hepatocellular injury (Fig. 7b) while PBS-treated SCID mice were completely resistant to ConA liver injury as assessed by routine histopathology and TUNEL staining as has previously been reported [21, 37]. Adoptive transfer of wild type splenocytes to SCID mice restored ConA-induced liver injury as assessed by histopathology and TUNEL staining (Fig. 7b) and serum transaminase levels (Fig. 7c). Interestingly, SCID mice reconstituted with PPARα−/− splenocytes showed substantial enhancements in serum ALT when compared to wild type mice reconstituted SCID mice given ConA (Fig. 7c). Consistent with the restoration of ConA-induced liver injury, SCID mice reconstituted with either wild type or PPARα−/− splenocytes had increased cytokine expression, specifically IFNγ and IL4 (Fig. 7d). Together, these data demonstrate the capacity of PPARα−/− splenocytes to reconstitute ConA liver injury and cytokine production to a level equal to or greater than wild type splenocytes. Further, these data suggest that deficiency in PPARα outside of the NKT population (i.e. Hepatocytes, Kupffer cells) are likely responsible for the reductions in NKT cell numbers in PPARα−/− livers.
PPARα-deficiency does not affect lipopolysaccharide-induced liver injury
Emerging evidence highlights the involvement of multiple cell populations during ConA-induced liver injury [38]. Specifically hepatic macrophages have been shown to contribute, at least in part, to pro-inflammatory cytokine expression and clotting factor production associated with tissue inflammation and necrosis [39]. Within the current paradigm, reduced NKT cell numbers and reduced IFNγ production correlate with reduced tissue injury in PPARα-deficient mice. The effect which PPARα-deficiency has on macrophage function within the liver has not been thoroughly investigated. To test their responsiveness, wild type and PPARα-deficient mice were administered lipopolysaccharide (5 mg/kg) by intraperitoneal injection 6 h prior to sacrifice. Serum and tissue were collected to assess liver damage and cytokine production. As shown in Fig. 8, LPS administration increased inflammatory cell infiltration as assessed by histopathology, liver injury as measured by serum ALT levels, and significantly increased serum IL12 levels as measured by ELISA. Loss of PPARα did not affect LPS-induced inflammatory cell infiltration or tissue injury but did lead to enhanced serum IL12 levels but reduced serum IFNγ protein. Together, these data suggest that hepatic macrophage function is similar between wild type and PPARα-deficient mice.
PPARα-deficiency reduces hepatic IL15 expression
NKT cell survival is dependent on the expression of key proteins and cell surface molecules [1, 4, 40]. Both hepatic CD1d expression and IL15 production within the liver have been associated with NKT cell survival. Loss of PPARα leads to a reduction in the number of hepatic NKT but the mechanism governing this effect remains unclear. To begin to understand the potential regulators of this response, RNA harvested from untreated wild type or PPARα-deficient mice were examined for the expression of both CD1d and IL15. As shown in Fig. 9, PPARα-deficient livers show reduced expression of IL15 but not CD1d. These data may provide the first mechanistic link between PPARα-deficiency and the absence of hepatic NKT cells.
T cell-dependent liver injury represents an important component of a number of liver pathologies including autoimmune and viral hepatitis [41–43]. Defining the mechanisms by which lymphocytes exert their damaging effects represents an important area of scientific investigation. To this end, the current series of studies have identified PPARα as a potential regulator of hepatic T cells. Specifically, data here demonstrate the importance of PPARα in the recruitment and/or survival of NKT cells, independent of its function within these cells. The ability of PPARα to regulate the immune cell composition of the liver and lymphocyte responses may have important clinical implications in the treatment of a number of T cell-dependent liver pathologies.
ConA-mediated liver injury is a well-described model of T cell dependent acute hepatitis in rodents [16]. NKT cells are activated by ConA in a CD1d-dependent manner to produce IFNγ and IL4 which serve to further activate this cell population as well as recruit and activate additional inflammatory cells including macrophages, thereby acting as a bridge between the innate and adaptive immune response [17–19, 24, 44, 45]. Recent studies by Li et al. as well as studies from our laboratory have drawn a strong correlation between the presence of hepatocellular lipid, absence or reduction in hepatic NKT cells and the production of a shifted Th1-type cytokine response in the liver [2, 46]. The results of the current study suggest that the loss of PPARα leads to a similar depletion of hepatic NKT cells which likely contributes to the reduced hepatocellular injury observed following both ConA administration as well as αGal treatment. Importantly, the reduced responsiveness to αGal supports the flow cytometric data indicating reduced NKT cells as numerous reports have shown a potential downregulation of defining cell surface markers, particularly NK1.1 and/or CD49b. Together, these data highlight the deficiency in NKT cells, both in phenotypic appearance and functionality in PPARα deficient mice, a key regulatory immune cell within the normal liver but stop short of defining the mediators of this hepatic immune phenotype.
Hepatic NKT cells are regulated by a variety of factors, both membrane bound as well as secreted. Loss of CD1d, reduced production of supportive cytokines such as IL15 or over-production of inflammatory mediators have all been associated with their depletion [45]. Likewise, activation itself may reduce NKT cell function, alter their cell surface phenotype, or induce cell death. Multiple models of fatty liver have shown interactions with many of these factors. Leptin-deficient ob/ob mice have reduced NKT cell numbers which correlates with reduced hepatic CD1d expression as well as blunted IL15 production [40, 47]. Loss of PPARα did lead to a small but significant reduction in tissue IL15 expression but had no effect on CD1d tissue expression suggesting that PPARα-deficiency, or the accumulation of hepatic lipid that occurs as a result, may influence hepatic production of this important supportive signal as has been noted in other models of fatty liver disease [1, 4, 40]. Choline-deficient diet feeding leads to a time-dependent increase in lipid accumulation and hepatic macrophage IL12 production which inversely correlates with NKT cell numbers [2]. Moreover, genetic deletion of IL12p40 restores the hepatic NKT cell population independent of changes in hepatosteatosis. Within the current study, loss of PPARα leads to a mild microvesicular lipid deposition which correlates with a small but significant increase in serum IL12 production at baseline (Fig. 8). Such data highlight a consistent IL12 response in the presence of excess hepatic lipid accumulation though the mechanism for this upregulation remains unclear. Previous studies reported the ability of PPARα activation to suppress NFκB activation in macrophages limiting their production of a number of inflammatory cytokines [9, 10]. Likewise, loss of PPARα interrupts normal lipid and cholesterol metabolism in macrophages similar to that seen in hepatocytes [48]. Altered lipid homeostasis can have a profound effect on macrophage function, promoting inflammatory cytokine production. Loss of fatty acid binding protein 5 (FABP5) promotes LPS-induced IL12 production in vitro and in vivo from hepatic macrophages further supporting an interaction among lipid, macrophages, and their production of IL12 [3]. The link between IL12 and PPARα at the level of the macrophage remains unclear but is likely related to lipid accumulation and subsequent inflammatory transcription factor activation.
The current series of studies are limited by the global loss of PPARα. Adoptive transfer experiments of lymphocyte populations allow for more selective examination of the effects of this transcription factor. Data from this approach support the notion that loss of PPARα leads to a hepatic microenvironment which is not conducive to NKT cell survival. Supporting this idea, reconstitution of PPARα sufficient, lymphocyte deficient SCID mice with either wild type or PPARα deficient lymphocytes restored ConA-induced tissue injury and cytokine production in these mice. In fact, reconstitution of SCID mice with PPARα-deficient lymphocytes caused a 4 fold enhancement in liver injury when compared to wild type lymphocyte reconstitution. The reasons for this enhancement in tissue injury are not clear. Previous studies have demonstrated the impact of PPARα deficiency on lymphocyte responsiveness [14, 15]. Loss of PPARα exaggerated IFNγ production by CD4+ T cells in vitro upon stimulation with CD3 and CD28. Pilot studies confirmed this enhancement in IFNγ production by PPARα deficient lymphocytes (data not shown). In vivo examination of IFNγ production did not, however, reveal significant increases in PPARα reconstituted mice when compared to wild type mice though IL4 levels were doubled. Further examination of the time-course of IFNγ expression is warranted in this setting to better understand its role though data from this study support a function for PPARα independent of the lymphocyte in the regulation of NKT cell function and ConA responsiveness.
Interestingly, in the current study, accumulation of lipid reduces NKT cell numbers and function but does not promote an enhanced Th1 response. This is in contrast to previous studies but may be related to the degree of lipid accumulation as well as the extent of NKT cell depletion. Indeed, previous studies have shown significantly higher levels of lipid accumulation as compared to the current results while also showing significantly higher numbers of hepatic NKT cells remaining following lipid accumulation [37, 40]. It may also be that PPARα regulates the function of other cells with respect to their ability to produce Th1-type cytokines. Data presented in Fig. 8 highlight the ability PPARα-deficiency to enhance lipopolysaccharide-induced IL12 production likely from macrophages but interestingly impair hepatic production of IFNγ. It is clear that macrophages contribute to ConA-induced liver injury as their depletion reduced hepatocellular injury in part through reductions in proinflammatory cytokine expression [49]. The involvement of macrophages in the current paradigm remains unclear and reduced IFNγ production following ConA exposure may result from impaired Kupffer cell responses. In vitro studies and αGal administration support a defective NKT cell response but further study is needed to determine the specific source(s) of Th1 cytokines in this and other models and the relative contribution of these cells to overall ConA-induced liver injury. It is clear that loss of PPARα leads to a significant reduction in hepatic NKT cell number and function and limits ConA-induced and αGal stimulated cytokine responses and associated tissue injury.
As discussed above, activation of PPARα stimulates peroxisome proliferation and transcription of a number of lipid metabolizing enzymes in rodents [7]. In humans, PPARα is present at low levels within the liver and does not appear to transactivate genes involved in peroxisomal β oxidation [50]. As such, chronic treatment with PPARα activators does not activate peroxisome or hepatocellular proliferation in humans as it does in rodents. Recent studies have demonstrated that activation of PPARα in human T lymphocytes results in strong reductions in the activation-induced expression of a number of cytokines including IFNγ, a finding consistent with its overall anti-inflammatory effects and its function in this immune cell population [51]. The role that PPARα plays in specific lymphocyte subpopulations as well as in tissue specific localization of these lymphocyte populations in humans has not, however, been explored. Given the results of the present study, modulation of PPARα function within the liver may indirectly modulate the immune response in humans. Additional investigation will be required to determine how PPARα affects lymphocyte function within the human liver.
In conclusion, data derived from the current series of studies demonstrates the importance of PPARα in the recruitment and/or survival of NKT cells within the liver. Consistent with these reductions in NKT cells, PPARα−/− mice shown strong resistance to ConA-activated and αGal stimulated cytokine production, specifically IFNγ, and subsequent liver damage. The role that other cell populations play in this process, particularly macrophages, cannot be fully addressed in the current paradigm. Further study is required to determine the exact mechanism by which PPARα regulates the localization and/or survival of NKT cells to the liver including the absolute importance of IL15 in this process and the direct contribution of macrophages both in NKT cell survival and tissue injury following ConA exposure. Understanding the mechanisms involved in PPARαdependent regulation of hepatic immune cell populations may prove useful in the design of therapies to modulate the immunological response of the liver.


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Andrew Siegel–Yahoo's head of corporate development, who is in charge of its mergers and acquisitions strategy–is leaving the company, according to sources.
The former General Electric exec, who came to the Internet giant just over a year ago, is well liked at Yahoo and also has been active in making the rounds in the tech scene in Silicon Valley.
Siegel's exit is part of a long line of departures of top talent under the leadership of CEO Carol Bartz.
Siegel has made some very prescient moves on game-changing companies for Yahoo to purchase–including Yelp, Foursquare and Groupon–but Yahoo has not been able to land them for a variety of reasons.
In Foursquare's case, the geolocation hot spot decided to remain independent and take more funding, after also talking to Facebook, and moneybags Google has hip checked Yahoo out of the running for social buying phenom Groupon.
Yelp, the reviews site that had unsuccessful talks with Google too, simply did not want to sell to Yahoo.
Both also were worried about Yahoo's future prospects and its top management's ability to turn the situation around, multiple sources confirm, as have other top start-ups that Siegel has been interested in.
And while he has bought some smaller companies for Yahoo–such as Citizen Sports and Associated Content–that's been frustrating to Siegel, said sources.
Sources also noted that Siegel has also been commuting to Yahoo's Sunnyvale, Calif., HQ from New York, where his family lives–also a factor in his decision to leave.
He apparently told his Yahoo bosses he was leaving just last week and the company has no replacement as yet.
Yahoo declined to comment about Siegel's status.

subset: commoncrawl


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Save The Mid has constantly questioned the creditability of the decisions made bu the Directors of the NHSCT, the decisions that were made in the name of patient safety have led to some of the worst episodes of patient safety within the trust.
A further review should be under taken by the DHSSPS and Heath Minister to investigate past decision made by Valerie Jackson.
The director was involved in the controversial decision to remove acute services from the Mid Ulster hospital, the closure at the time was classed a crisis closure. What needs to be answered who caused the crisis closure ?
Save The Mid will keep constant pressure upon the board of the NHSCT with the view that Chairman Jim Stewart be brought to Health Minister Edwin Poot's and questioned on the activity of the NHSCT board during his rein as chairman.
Valerie Jackson took up the post of Director of Acute Hospital Services on 2 November 2009. She trained as an RGN in the Royal Victoria Hospital (1985-1988) gaining her midwifery qualification in 1991. Having held many nursing posts in a wide range of hospital and community settings, she joined the Northern Ireland General Management Training Scheme in 1995 which involved working in range of posts in a variety of Trusts in Northern Ireland. She was appointed Project Manager for Phase 1 of the Royal Belfast Hospital for Sick Children in 1997.
Between 1998 and 2003 Valerie worked as Directorate Manager for Medicine, General Manager for Emergency Care and Divisional Manager for the Division of Medicine and Surgery before leaving the Royal Victoria Hospital in 2003 to take up post as Director of Nursing and Primary Care in the Ulster Community and Hospitals Trust.
In 2007 she was appointed to the position as Director of Older People Medicine and Surgery in the Belfast Trust and was also Executive Director for Nursing until June 2009 when she stood down from that role to allow Trauma and Orthopaedic services to join the Older People Medicine and Surgery Service Group.
- Accident and Emergency
- Acute corporate services
- Acute facilities
- AHP physiotherapy and dietetics
- Anaesthetics
- Cancer
- Critical care (intensive care and high dependency)
- Dermatology
- Endoscopy
- Gynaecology
- Integrated Clinical Assessment and Treatment Services (ICATS)
- Obstetrics
- Oncology
- Orthopaedics
- Outpatients
- Relevant outpatients
- Sexual health
- Surgical services
- Theatres
- Trauma


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Defense Date
Document Type
Degree Name
Master of Science
First Advisor
Albert Farrell
Second Advisor
Terri Sullivan
Third Advisor
Thomas Farmer
Peer victimization has been shown to negatively impact youth functioning and may be especially damaging during adolescence, given the increased importance of peers. However, there is a dearth of longitudinal research examining trauma symptomatology as an outcome of peer victimization with low-income, ethnic minority adolescents. The present study investigated this relation in a predominantly African American sample of 684 students assessed at five time points between the fall of their sixth grade and seventh grade school years. Growth mixture models grouped participants with similar victimization trajectories, and latent growth models related growth trajectories of physical and relational victimization to changes in trauma symptoms. Although initial levels of victimization were unrelated to changes in trauma symptoms over time, increasing victimization was associated with increasing trauma symptoms. These findings provide insight into the relation between peer victimization and trauma in an underserved sample of adolescents, with important implications for prevention efforts.
© The Author
Is Part Of
VCU University Archives
Is Part Of
VCU Theses and Dissertations
Date of Submission


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Many people these days are pointing to Finland as the world's top success story in student achievement. So what's their secret?
In the latest issue of NEA Today magazine, we feature an excerpt from a book by Stanford University scholar Linda Darling-Hammond that tells the Finnish story. Basically, Darling-Hammond explains, Finland did the opposite of what we're doing in America.
In the 1970s, reports Darling-Hammond, Finland's student achievement was low. But in the decades since, they have steadily upgraded their education system until now they've reached the top.
What's more, they took what was once a wide achievement gap between rich and poor, and reduced it until it's now smaller than in nearly all other wealthy nations.
Here's how:
* They got rid of the mandated standardized testing that used to tie teachers' hands.
* They provide social supports for students including a free daily meal and free health care.
* They upgraded the teaching profession. Teachers now take a three-year graduate school preparation program, free and with a stipend for living expenses. In Finland, you don't go into debt to become a teacher.
* The stress on top-quality teaching continues after teachers walk into their schools. Teachers spend nearly half of their time in school in high-level professional development, collaborative planning, and working with parents.
These changes have attracted more people to the teaching profession — so many that only 15 percent of applicants are accepted.
The Finns trust their teachers, Darling-Hammond reports. They used to have prescriptive curriculum guides running over 700 pages. Now the national math curriculum is under 10 pages.
With the support of the knowledge-based business community (think Nokia), Finnish schools focus on 21st century skills like creative problem-solving, not test prep.


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North American Network Operators Group
Date Prev | Date Next | Date Index | Thread Index | Author Index | Historical
Re: Service Provider Exchange requirements
> > If your switch is MCAST aware, you should be able to keep mcast traffic on > ports tagged for it to begin with. If your switch isn't mcast aware. you > need to find a new switch. MCAST aware means different things in different environments. Ideal is a switch that knows which multicast groups a particular port has joins on, rather than simply whether or not it is getting multicast traffic. In an ethernet fabric used as an exchange point, you have inter-AS multicast traffic, so sniffing IGMP doesn't do any good. Sniffing PIM sparse mode for joins would work. In some environments, you might be able to use RGMP to tell the switch which groups have been joined on a particular port (John Meylor mentioned over beer at the Cogent social that his group might consider writing up RGMP as an informational draft, so that number of enviornments may go up). > > As for jumbo frames, will someone remind me what the benefit of using a > larger MTU on the edges than you have in the core is? Is the edge device > going to aggregate 6 1500-byte packets into a single 9000-byte jumbo frame > for me?n > If it is not clear, I am talking about using jumbo frames on ethernet VLAN used in an exchange point; this would provide a migration path for service providers who have jumbo frames to the edge, because they could trade them over the exchange point frabric. They could, of course, do the same thing over a private interconnection. regards, Ted Hardie


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1. Meteor. The largest, with seating in three sections (front, middle, rear). The front has a panoramic view of the river ahead. The middle is the largest section and has a kiosk that provides hot water, beer, and candy. There is an open air walkway between the middle and rear. The rear section is the warmest and noisiest because it is close to the engines.
2. Voskhod. Narrower than a Meteor, with just two sections. Front section also provides panoramic views.
3. Raketa. Just one passenger section. No wrap-around windows up front.
4. Polsye. Roughly the same size as the Raketa, but no front view (due to bridge) or rear view (due to engine).
5. Zarya. Not technically a hydrofoil, as it does not have underwater wings. But it does have water propulsion. Flat-bottomed for shallow waters. It will run right up to the riverbank without the need for a gangway or pier. Inside is like a local bus.


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Bereavement Classes to begin Feb. 4
Sangre de Cristo Hospice Bereavement Classes will begin from 5:30-7:30 p.m. Feb. 4 and run for five consecutive Tuesdays, ending March 4.
This group of classes was developed to support anyone who has experienced the loss of a loved one. The curriculum will help participants share their experiences and increase their knowledge of the bereavement process. These classes are free and open to anyone in the community who is struggling with the loss of a loved one.
To sign up for this group, call Jenny Lehl at 275-1261.
Crime Prevention Tips from the CCPD
Opportunity offenders are generally unskilled at criminal activity and often under the influence of drugs or alcohol at the time of the crime.
An endless number of crimes are committed by the opportunity offender and they can be very serious in nature. They look for cars with unlocked doors or keys in the ignition to rob or steal. They watch you pull out a wallet full of money and credit cards during a purchase. They see a front door that someone forgot to close or lock. They notice that you are under the influence and thus vulnerable.
Most opportunity offenders could not pick a lock if their lives depended on it. They commit a burglary by going through unlocked doors or windows, or they kick a door down or smash a window.
The most important thing to understand about protecting your home is that "target hardening" is one of "the" most successful forms of crime prevention. The bottom line is that opportunity offenders (and many more determined offenders) will be persuaded to look elsewhere or give up entirely when they realize that breaking into your home is not worth the effort. They only have the skills to take advantage of what you make easy for them.
The Cañon City Police Department is happy to discuss your personal and home security concerns with you. We also provide security surveys and will make recommendations if any issues are identified. Do not hesitate to call us regardless of the topic.
If we cannot find an answer for you, we will research the topic and come back to you with suggestions. Those with concerns regarding Crime Prevention, safety in their home or neighborhood, call (719) 276-5600, ext. 3917 to speak with Jen O'Connor, Crime Prevention coordinator, or email email@example.com. Be safe.
Daily Record Staff


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Madonna Fights Back: Inside Rolling Stone's New Issue
26/02/2015 06:10
Madonna gets fiery and introspective, discussing ageism, Gaga and more in our cover story
"This is what a 56-year-old ass looks like!" Madonna tells Rolling Stone, discussing ageism in pop culture.
Madonna returns to the cover of Rolling Stone in our latest issue (on stands Friday) giving her most revealing, introspective and fiery interview in years. In the in-depth Q&A with senior writer Brian Hiatt, Madonna discusses her real feelings about Lady Gaga, her marriage to Guy Ritchie, her relationship with Judaism, her assessment of Kanye West, her love for Whiplash and much more.
"I don't think she wants my crown," Madonna says, referring to Gaga. "We live in a world where people like to pit women against each other. And this is why I love the idea of embracing other females who are doing what I'm doing. . .The only time I ever criticized Lady Gaga was when I felt like she blatantly ripped off one of my songs. It's got nothing to do with 'she's taking my crown' or 'she's in some space of mine.' She has her thing. I do think she's a very talented singer and songwriter. It was just that one issue. And everybody's obviously running with it and turned it into a huge feud, which I think is really boring, quite frankly. And you know what? I don't care anymore. Here's the thing: one day everyone's going to shut up about it. You'll see! I have a plan. "
But Madonna, who will release her new album Rebel Heart on March 6th (read our review of the album here), reserves her most passionate and eloquent remarks for the topic of ageism, in pop writing and in society. "It's still the one area where you can totally discriminate against somebody," she says, "and talk shit. Because of their age. Only females, though. Not males. So in that respect we still live in a very sexist society."
"No one would dare to say a degrading remark about being black or dare to say a degrading remark on Instagram about someone being gay," Madonna continues. "But my age – anybody and everybody would say something degrading to me. And I always think to myself, why is that accepted? What's the difference between that and racism, or any discrimination? They're judging me by my age. I don't understand. I'm trying to get my head around it. Because women, generally, when they reach a certain age, have accepted that they're not allowed to behave a certain way. But I don't follow the rules. I never did, and I'm not going to start."
And if you're wondering if there was a message behind showing off her bare butt on the Grammys' red carpet: "This is what a 56-year-old ass looks like, motherfuckers!" she says. And to the suggestion that her awe-inspiring physique isn't exactly average, she retorts, "You know what? It could be the average some day! That's the thing." (Listen to Madonna's entire statement on ageism in pop culture below.)
"When I did my sex book, it wasn't the average," Madonna says. "When I performed 'Like a Virgin' on the MTV Awards and my dress went up and my ass was showing, it was considered a total scandal. It was never the average, and now it's the average. When I did Truth or Dare and the cameras followed me around, it was not the average. So if I have to be the person who opens the door for women to believe and understand and embrace the idea that they can be sexual and look good and be as relevant in their fifties or their sixties or whatever as they were in their twenties, then so be it."


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Atomic Improv
Atomic Improv stars three-time world improvisational champion Donovan Workun. Workun has performed thousands of shows all across North America and Europe. He has shared the stage with some of the best improv comedians in the world including Wayne Brady and Colin Mochrie from Whose Line is it Anyway, and Second City alumnus Dave Thomas, Joe Flaherty, and Mike Myers. You can hear Donovan weekly on CBC radio 1 on the national comedy series The Irrelevant Show, and he appears regularly on your TV selling everything from pizza and carwashes, to banking and furniture.


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Tiger Woods is going to donate $1 million dollar to charity if nobody asks about his injured leg.
Tiger is attending a news conference for the AT&T National golf tourney in Philadelphia and he made this announcement via Twitter.
Prince William and Kate Middleton had a visit with President Barack Obama and First Lady Michelle Obama at Buckingham Palace on Tuesday. The royal newlyweds have been married for almost a month.
Leo DiCaprio's ex-girlfriend Bar Refaeli showed that she still hot in a bikini. She proved that the couple's breakup did not hurt her amazing body.
Ryan Seacrest's girlfriend Julianne Hough is co-starring in the upcoming Tom Cruise flick "Rock of Ages" She filmed a passionate scene with Diego Boneta in which the couple locked lips in Florida yesterday.
Michael Lohan pleaded not guilty yesterday to a misdemeanor charge of inflicting injury on a cohabitant. Lohan was arrested March 21st after an argument with his ex Kate Major turned violent. He stills claims he did not touch her and in fact claims she is the one who attacked him.
Justin Timberlake attended an SNL after party in NYC, where he was spotted hanging out with his rumored new lady friend Olivia Wilde.


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Sapele Mahogany was used to make the main deck and birch for the bottom two layers. The holes and shape was created using a CNC router system.
The process required to make this boat takes a few extra steps, but the wait is well worth the finished product.
Hand rubbed polyurethane was use to coat and protect it.
|Finished boat top view.
|Finished boat side view.
|Out of glueup and shaped by the bandsaw..starboard side
|Out of glue up and shaped by the bandsaw ..port side
|Just out of glue up
|Boat in glue up
|All three levels ready for glue up
|CNC drilling the peg and card pockets
|CNC drilling the holes
|Photo with sailboat name example.
Stay tuned for more updates soon......:-) ****************************************************************
For more information about ordering your own custom board go to: http://www.splinterswoodworks.com/


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Barack Obama has apparently committed blasphemy. In an interview in Florida on Sept. 29, he dared to say that America had gotten "soft." The denunciations have come in fast and furious from the right. Mitt Romney, Rick Perry, Eric Cantor and the Wall Street Journal editorial page are all shocked--shocked--that the President could say such a thing. "America is the greatest nation in the world," Cantor declared. Romney concocted a confusing metaphor about America carrying Obama on its shoulders, but his basic point was the same. Now, if you watch the clip, here's what the President said: "The way I think about it is, you know, this is a great, great country that had gotten a little soft, and we didn't have that same competitive edge that we needed over the last couple of decades. We need to get back on track." Isn't this self-evidently true? Isn't this what conservatives have been saying for decades?
The evidence on the topic is pretty clear. The U.S. is slipping, by most measures of global competitiveness. It has dipped slightly in the World Economic Forum's (WEF) rankings to No. 5, behind Sweden, Singapore, Finland and Switzerland. But the WEF rankings are based, in good measure, on surveys--polls of CEOs and the like. Other studies, using hard data, show America slipping further behind. The Information Technology and Innovation Foundation finds that in category after category--actual venture-capital funding, research and development--the U.S. has dropped well behind countries like Japan, South Korea and Sweden. The foundation measures 44 countries and regions on their efforts to improve their competitiveness over the past decade. The U.S. comes in next to last.
Perhaps the most crucial measure of our ability to compete in a global economy is our educational attainment, especially in science, math and engineering. A generation ago, America had the highest percentage of college graduates in the world. Today we're ninth and falling. The WEF report ranks the U.S. a stunning 51st in science and math education. If a willingness to study science, math and engineering is an indication of being willing to work at hard stuff, there is no question that we are going soft. In 2004 only 6% of U.S. degrees were awarded in engineering, half the average for rich countries. In Japan it's 20%, and in Germany it's 16%. In 2008--09 there were more psychology majors than engineering majors in America and more fitness-studies majors than physical-sciences majors.


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One of the best ways to get people to achieve excellence is to inspire them through your own example. No matter how much you talk or what you do, if you aren't stretching and growing, if you aren't giving until it hurts, forget about asking them to do so.
Most Olympic athletes got to where they are because they were inspired by an Olympic athlete who preceded them. Few of us, if any, have the capacity to be self-inspired. We need another person to help light the flame. You are their role model. How your people perform will always reflect to some degree the behavior and ideals you live out day-to-day basis.
Virtually everything you do in some way implies a standard to your people—the way you dress, the way you interact with peers and superiors, whether or not you start your meetings on time, the quality of your work. It is a good idea to do a quick check on yourself from time to time to see if you are performing the way you want your people to perform.
My Consultancy–Asif J. Mir – Management Consultant–transforms organizations where people have the freedom to be creative, a place that brings out the best in everybody–an open, fair place where people have a sense that what they do matters. For details please visit www.asifjmir.com, and my Lectures.


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Dogs generally experience pain for many of the same reasons that we do: dental issues, arthritis, physical injuries, or just not feeling well in general. But dogs cannot speak human, so having the ability to read your pooch and see when something is not right is an important to ensuring his health and quality of life.
Being able to tell if your pooch is feeling pain is crucial in identifying the cause and allowing you to respond immediately to find the appropriate treatment.
You'll know something may be wrong if…
- Your dog cries out in pain. Vocalization is one of the closest things that dogs have to speaking. It's one of their most obvious ways to communicate that something hurts. If your pooch whines and whimpers for no clear reason, this can be a sign that he is in pain.
- Your dog starts limping. This is another rather straightforward symptom that you should watch out for. Dogs maybe hobbling or walking lamely for various reasons, usually all of them associated with some sort of discomfort. Limping can be the result of a wide variety of issues ranging from arthritis or joint pain, to back or knee injury, to pain in the foot or paw pad.
- Your dog pants excessively. Because panting is a very normal thing for pooches, this particular indicator of pain can be rather tricky. When your dog pants at unusual times, such as the middle of the night, or when he's been resting and shouldn't be out of breath or thirsty, especially if the panting is associated with trembling, there could be a real problem.
- Your dog licks a localized spot too much. Most dogs lick their throbbing wounds, stinging paw pads, tender broken toe nails, and other parts of their body when they are in pain. Although it doesn't always suggest soreness, you can tell if something is wrong when you catch your furball licking and chewing a certain spot on his body over and over again.
- Your dog is hesitant to get up or sleeps more than usual. Dogs have a natural, instinctual tendency to hide their pain. As pet parents, it's important to make note of even the most subtle changes in our dog's typical behavior. Of course, if a dog is suddenly very slow getting up or hesitates to walk, it becomes clear fairly quickly that something is wrong. Other dogs, however, may only slow down slightly, or may simply appear to me sleepier than normal.
- Your dog's temperament and behavior suddenly changes. Like us, dogs can also get a little irritable when they aren't feeling well. A dog that is normally pleasant and friendly could become aggressive without warning when he's in pain. He may even try to bite, especially if the area that hurts is touched. Other pooches may all of a sudden appear more needy and may seek more attention from people than usual. Loss of appetite may also be observable.
If you think Fido is not feeling well, a trip to your veterinarian may be necessary. There are many pain relief options available for dogs nowadays, but before trying any of them, make sure that you discuss the situation with your vet.
Never, ever give your pooch over-the-counter medications like Ibuprofen and Tylenol as these drugs are very toxic to dogs.


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- Hall rental = $25.00** per hour with a 2 hour minimum. / Non Resident Rate = $50.00
- Kitchen = $75.00** (if required) / Non Resident Rate = $150.00
- Non-Refundable $25 Fee for all rentals
A security deposit of $150.00 is refundable after inspection of rental areas.
Renters must present proof of personal injury coverage in excess of $200,000.
Facilities are not available to rent for profit generating functions.
Alcoholic Beverages are not permitted on the premises.
All rental requests must be approved by the department at a regular monthly meeting.
Monthly Meetings are held on the 1st Monday of the Month.
A member of the department must volunteer to sponsor the function for the request to be granted.
Click here to download the rental agreement.
For a prompt response email the President at: email@example.com
If you don't have access to email please leave a message on the Presidents cell phone518-451-6648 and they will return your call at their earliest convenience.


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In a single year, the price of Bitcoin grew by a factor of 56. Imagine that happening to $100 in your checking account — you'd get $5,600.
It's a neat stat that we at VentureBeat didn't come up with on our own. It's one of the many nifty bits of information tucked inside a 91-page report that Bitcoin news outlet CoinDesk issued today.
The report does a great job of stepping back from the incremental news of the digital currency and displaying its bigger trends. It also picks out many of the winners in the Bitcoin game. Here we deliver a few of the most fascinating findings from the report, along with some notes of our own.
For starters, Bitcoin's "market cap" at the end of 2013 was $9.2 billion, according to the report. CoinDesk performs that calculation by multiplying the price of Bitcoin by the number of Bitcoins outstanding. It's more a representation of the currency's value than the market cap of a public company, which is determined by multiplying current share price by number of shares outstanding. Still, the Bitcoin "market cap" number hints at the financial heft that was hardly at the beginning of 2013.
More than $200 million has been spent on Bitcoin mining equipment, the report states.
Venture capitalists poured more than $98 million into cryptocurrency startups, according to the report. Around 70 percent of that investing took place in the United States. Beneficiaries of this funding include BitPay, Circle, Coinbase, itBit, and OpenCoin.
Payment processors have received $36.7 million, which is more venture funding than other kinds of startups in the Bitcoin ecosystem, like exchanges and mining-hardware makers.
It's also interesting to note that CoinDesk's $98 million venture capital figure is out of sync with the latest total from Venture Scanner, $83.7 million. It points to how fast the Bitcoin ecosystem is advancing.
The report notes that early on, people viewed Bitcoin as an asset that one invests in, like gold. It has since evolved into something businesses want in on and consumers want to pay with. All the while, there's been a correlation between Bitcoin's price fluctuation and the number of times that the media mentions it.
CoinDesk wagers that later this year, Bitcoin will get out in front of more consumers as merchants continue to adopt it and more Bitcoin conferences will be held. But the company also expects that digital currencies other than Bitcoin, like Litecoin and Peercoin, will garner more interest. That's not hard to believe, with Bitcoin's price on a general decline since the beginning of this year, and with key exchange Mt. Gox shutting down transactions — a fact not mentioned in the report.
Even so, CoinDesk sees more funding and acquisition activity in store for Bitcoin startups this year. Perhaps such deals will involve the top Bitcoin wallet companies, which are Blockchain, Coinbase, Bitcoin Wallet, and Mycelium.
If you're interested in a detailed list of Bitcoin startups that have received investments, an overview of challenges and opportunities facing Bitcoin, and other information, check out the full report here:
And to learn even more about Bitcoin, read our introductory guide to the digital currency.


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Love getting a great deal on crafting supplies to add to your stash? For Black Friday this year, you'll have the opportunity to get amazing crafting supplies at an amazing price with Close To My Heart (CTMH)!
For 24 hours only, we'll feature themed mystery boxes filled with retired Close To My Heart products, all at a major discount! These mystery boxes will have five different price points ranging from $35 USD/$55 CAD to $150 USD/$160 CAD, with some mystery boxes being offered at 50% off the retail value! All mystery boxes will be available beginning November 25, 2016 at 12:00 am (MST) while supplies last—so grab a box for your stash before they're gone!
You may receive duplicate items in the mystery boxes. Many of the items included in the mystery boxes are considered consumable, which just means that having more than one would be even better! In the instances that duplicates are included, we won't put more than two identical items in a single mystery box.
What are the prices for the mystery boxes?
The mystery boxes will be made available in five tiers. Some boxes may contain items included in other mystery boxes, at other price points. The price tiers are as follows:
• $35 USD/$55 CAD mystery boxes (on average, a $70 USD/$85 CAD value!)
• $50 USD/$65 CAD mystery boxes (on average, a $90 USD/$105 CAD value!)
• $75 USD/$85 CAD mystery boxes (on average, a $140 USD/$165 CAD value!)
• $115 USD/$125 CAD mystery boxes (on average, a $235 USD/$280 CAD value!)
• $150 USD/$160 CAD mystery boxes (on average, a $335 USD/$395 CAD value!)
How long will each mystery box be available?
Each mystery box will be available for up to 24 hours, while supplies last. Visit my Online STORE early for the most options!
Thanks for reading and get your stash on Friday!!!


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Enclosed tuning machines with a 12-to-1 gear ratio will keep you in tune. No string slipping with these bad boys.
Choose gold, chrome or black.
Dixie Bridge
Custom Made Top Guitar bridges feature adjustable saddles and through body string suspension.
Reedsdale Bridge and Tail Piece
The Reedsdale requires a bridge and a tail piece. We use a tune-o-matic bridge that allows you to fine tune your guitar's action to fit your playing style.
Available in gold, chrome and black.
Want to add something really special to your guitar? The famous Bigsby was the first successful design of what is now known as the whammy bar or tremolo arm. The Bigsby is installed on the top of the guitar. When the arm is pushed down, the strings loosen, lowering their pitch. When the arm is released the strings return to their neutral position.
Switches, Neck Plates, etc.
All of our switches, neck plates, jack plates and screws are available in gold, chrome and black.
We currently offer 22 different paint colors and stain finish options.
The paint we use is part of the PPG Envirobase line. This is a waterbased paint that contains very low VOC levels and therefore is some of the most environmentally safe paint used on any guitar. This same line of paint has been adopted by high end car manufacturers such as Lamborghini and Ferrari. This paint is also an important element in our production process because it dries quickly, allowing us to build our guitars in less time.Metallic Paint
If you want to add a bit of sparkle to your paint, choose a metallic color. These colors have a small flake in the paint that shines when hit with light. We offer metallics in the following colors:
Black, Silver, and Gold
UV Clearcoat
After a guitar has been painted or stained, it is sealed in a protective clearcoat which prevents the paint from chipping and also gives a guitar its shiny finish. Typically clearcoat takes a long time to dry. However, at Custom Made Top Guitar, we use a special clear coat that can be fully hardened quickly with the use of a high powered ultaviolet (UV) light. This clearcoat levels beautifully giving your guitar an almost mirror-like finish.
Text and Graphics
Designing with Text and Graphics
There's no better way to customize your Custom Made Top Guitar than by adding graphics and custom text. To get started, just look for the "Paint Upgrades" category on any of the guitar configurator pages. Then choose the "text" or "graphics" subcategories. The video below shows you exactly where to find these options.
One of the more popular areas of the guitar to add custom text is the headstock. Typically, guitar manufacturers put their brand name on the guitar's headstock. With Custom Made Top Guitar you can add your own name, logo, or other text to the headstock and brand it anyway you want. Or you can leave it blank.
How We Apply Text and Graphics
The process for applying text and graphics to your guitar starts with graphic design software. Our team creates a vector graphic and then prints a stencil using a vinyl plotter. This stencil is then applied directly to the guitar. Next the guitar goes to the paint booth with the stencil attached and paint is sprayed. The stencil is then removed leaving behind your text or graphic.
Adding Graphics/Text To Your Pickguard
Text, graphics and even full-color photos can be printed onto our pickguards using a high resolution printer. We print on the back of a clear acrylic pickguard which, when attached to the guitar, prevents the image from being scratched off by a pick. The acrylic gives the pickguards a nice glossy finish.
After your guitar has been custom painted to match your design, it is assembled by hand in our shop in Austin, TX.
Our bolt-on neck is set using four 1-1/4" screws using our proprietary neck setting jig to ensure that it is perfectly straight with respect to the body. Next, the bridge is set and the tuners are installed.
The inside of the control cavity is coated with shielding paint to eliminate any interference from other electronics. The pots, switch and pickups are then installed and hand wired. The fretboard is then treated a special conditioning oil to enhance character and protect the wood's natural moisture content.
Once the remaining components are assembled, we string up your guitar and begin adjusting the bridge to set the action. Bridge height and saddle location are meticulously set to give your guitar great feeling action and perfect intonation. Your guitar is inspected and tested by our quality control team before being securely packaged and shipped.


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North Korea's leader Kim Jong-il appeared anxious to show China and the US that he's open to dialogue as the reclusive North edged closer Friday to six-party talks on its nuclear weapons.
North Korea's nuclear envoy, Kim Kye-gwan, returned from Beijing on Friday just as a top UN political official concluded a four-day visit to Pyongyang. Both missions look to analysts as unmistakable moves to resume six-party talks not held since December 2008.
"It looks like North Korea is paving the way to return to talks," says Han Sung-joo, who has served as South Korea's foreign minister and ambassador to Washington. "They want to do a favor to China, and they want to placate both the US and South Korea."
The most obvious sign of that willingness were reports here that envoy Kim Kye-gwan is scheduled to fly next month to Washington, where he is expected to meet the US envoy on Korea, Stephen Bosworth, and other top officials.
South Korea's Yonhap news agency quoted one source as saying the date for Mr. Kim's trip "has already been set" though there was no confirmation from Washington. Kim in Beijing conferred at length with China's newly appointed envoy on Korea, Wu Dawei, a former vice foreign minister whose position seems analogous to that of Mr. Bosworth.
At the same time, Lynn Pascoe, UN undersecretary-general for political affairs, met North Korea's titular leader, Kim Yong-nam, passing on what Pyongyang's Korean Central News Agency called "a verbal personal message and a gift from the UN secretary-general," Ban Ki-moon.
Neither the nature of the gift nor the contents of the message were disclosed, but Mr. Pascoe's mission appears as part of a drive to get North Korea to return to talks. The most critical sticking point is North Korea's demand for lifting stringent economic sanctions imposed by the UN Security Council after the North test-fired a long-range missile last April and then detonated an underground nuclear device in May.
After the meeting, Pascoe said the North had not rejected a return to the negotiating table, but Pyongyang is reluctant and holding out for the removal of the sanctions.
"They're certainly not eager, not ruling out, but not eager to return to six-party talks," he told reporters in Beijing.
As pressure builds up for talks on the North's nukes, the foreign ministers of Japan and South Korea held lengthy meetings this week at which they showed unusual rapport on dealing with North Korea and agreed on the need to maintain sanctions.
South Korea's foreign minister, Yu Myung-hwan, and Japan's foreign minister Katsuya Okada, promised to put teeth into their words by sharing intelligence – an unusual step considering the history of animosity between Korea and Japan – after Mr. Okada apologized profusely for Japan's annexation of Korea 100 years ago this year. Okada was quoted by a spokesman for South Korea's President Lee Myung-bak as having remarked, "This year is a sensitive year" in view of the anniversary but that Japan's Prime Minister Hatoyama wants "future-oriented" relations.
"There is fairly good coordination between the two countries on the nuclear issue," says Mr. Han, South Korea's former foreign minister. "It's to the credit of both the Japanese and Korean governments and the leaders of both countries."
Lim Jung Taek, foreign press director at South Korea's foreign ministry, denies that close ties between South Korea and Japan are aimed against China, on which North Korea relies for military and economic survival.
"You can say we were aligning against North Korea," he says, "but not against China" – which he credits with playing a major role in trying to get North Korea back to six-party talks.


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tag:blogger.com,1999:blog-6371386314838781559.post8443644119226622873..comments2020-08-07T06:03:53.115-07:00Comments on Anktangle: SPD? and Dairy-FreeAnonymoushttp://email@example.com:blogger.com,1999:blog-6371386314838781559.post-90803796355626823902011-09-27T22:12:55.456-07:002011-09-27T22:12:55.456-07:00So, I'm obviously coming in on this super late...So, I'm obviously coming in on this super late in the game here and I don't know what has progressed since you originally wrote this post. But I've been just kind of jumping around on your blog and getting to know you, this awesome gal whose milk my little one is gonna be snacking on soon! ;)
I just wanted to give you a little encouragement and say that our older son, Charlie, actually has SPD (was officially diagnosed at about 4 years old, I think). And while can be difficult at times, he's such an amazing, unique kiddo. And I really feel like the SPD, while it can be hard to deal with, really also adds a lot to his "whole package" and his uniqueness. I really wouldn't take it away from him, if doing so would change him at all. I really wouldn't. He's so sensitive, and smart, and perceptive, and just plain awesome. And in a strange way, I credit some of this to his "hyper" senses. :)
Again, I don't know what's down the road for Daniel, or if he'll ever be diagnosed (or maybe already has been?). But I just want to tell you that it's going to be okay, you will work through these things with your little guy, and learn so much from the challenges and you will be so proud as you watch him overcome some of his obstacles. Hang in there, I know it's not easy, but it does get easier, I promise.Marshttps://email@example.com:blogger.com,1999:blog-6371386314838781559.post-1497902986899685512011-01-01T14:50:28.941-08:002011-01-01T14:50:28.941-08:00@Kelly: Thank you so much for your empathy. Daniel...@Kelly: Thank you so much for your empathy. Daniel (like your baby, it sounds) will cry harder and harder if we leave him even for a moment when he's upset.

Thanks again for letting me know about your experiences and helping me to have some perspective about what we're going through. =)Anonymoushttps://email@example.com:blogger.com,1999:blog-6371386314838781559.post-46423697241673288802011-01-01T12:39:27.963-08:002011-01-01T12:39:27.963-08:00My first daughter cried SO very much in the first ...My first daughter cried SO very much in the first several months of life, and like your Daniel, seemingly strange things would help comfort her - like putting her in her carseat & lifting her straight up/down or playing music with a loud bass line...
The only advice we got from our pediatrician was "you have to just put her down; don't always nurse when she cries; let her cry to sleep". Sigh. Our baby was a baby who would only cry MORE and HARDER and LONGER. No advice would've been better than that "advice".
Eventually, she started to let up, and around 9/10 months, I started an elimination diet on myself (and she went back to breastmilk only) when we switched pediatricians (thankfully) and we did see a difference (in her eczema too).

But even now at age 6, she's INTENSE. And brilliant and empathetic and intuitive, but seriously intense.
Some babies, some children, just are. And its okay. I wrote a post about our experience with our "high needs" baby - if you're interested. http://www.kellynaturally.com/post/Why-High-Need-Instead-of-Fussy.aspx
((hugs)) It DOES get better.kelly @kellynaturallyhttp://email@example.com:blogger.com,1999:blog-6371386314838781559.post-49991610471113715472010-12-10T18:09:01.524-08:002010-12-10T18:09:01.524-08:00I just had a moment to watch the video (I read the...I just had a moment to watch the video (I read the post the day you posted it, of course) and Mini Jaymz' little sounds while he gets massaged are ADORABLE.

I wish I could do something to make Daniel cry less. But like we've talked about a million times, when that boy can talk, he will be a hilarious little firecracker. And maybe he'll explain some of why he's been such a fussy little babe.Rachel @ Lautaret Bohemiethttps://email@example.com:blogger.com,1999:blog-6371386314838781559.post-28272209203289638232010-12-08T21:07:49.425-08:002010-12-08T21:07:49.425-08:00Thank you, Rena. You are so kind. I appreciate you...Thank you, Rena. You are so kind. I appreciate your prayers very much.Anonymoushttps://firstname.lastname@example.com:blogger.com,1999:blog-6371386314838781559.post-53632299215441394972010-12-08T20:42:26.346-08:002010-12-08T20:42:26.346-08:00Wow. I can't imagine how you do it, Amy. I kno...Wow. I can't imagine how you do it, Amy. I know it must be absolutely exhausting. I'll keep your little family in my prayers.Unknownhttps://email@example.com:blogger.com,1999:blog-6371386314838781559.post-40769308083058508932010-12-08T16:52:48.863-08:002010-12-08T16:52:48.863-08:00@Kristen: Thank you for telling me your story. It&...@Kristen: Thank you for telling me your story. It's nice to know the dietary restrictions get easier as things progress, and that this could be the solution to all the crying!Anonymoushttps://email@example.com:blogger.com,1999:blog-6371386314838781559.post-91607716335452479652010-12-07T15:35:02.424-08:002010-12-07T15:35:02.424-08:00I hope that you can find something that does the t...I hope that you can find something that does the trick with helping him.

My son is almost nine months old and he was screaming pretty much every waking moment when I went on an elimination diet. I'm gluten-, dairy- and soy-free. It was hard at first, but has gotten easier. The nice thing is that there is quite a bit of gluten-free/casein-free food on the market. Please email if you want or need any tips.

Best of luck!Kristen @ My Semi-Crunchy Lifehttps://email@example.com


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Tonight"s 12-12-12 Sandy benefit Concert in New York will be sure to be full of fun collaborations and surprises. Among the artists performing are Bruce Springsteen, Paul McCartney, Lady Gaga, Kanye West, The Rolling Stones, Bon Jovi, Alicia Keys and The Who. In addition to some of the rumors already out there, we chatted with some of our sources for some other moments to keep your eyes open for at Madison Square Garden.
Here are seven things to watch for:
*Paul McCartney performing with Nirvana"s Dave Grohl and Krist Novoselic. British tabloid The Sun broke this story and ran a hilarious photoshopped pic of Sir Paul in a Nirvana photo sporting Kurt Cobain"s hair. They will reportedly play a new song. McCartney and Grohl played together at this year"s Grammys, but we have no idea on if this rumor is true, and, quite frankly, we"d rather see the trio play some classic rather than a new song (though certainly not a Nirvana classic). We reached out to Grohl"s publicist who would neither "confirm nor deny" the rumor and cheekily added, "You"ll have to wait and see tonight."
*Pearl Jam"s Eddie Vedder and Pink Floyd"s Roger Waters. Waters spilled the beans on his performance last night on "Last Night With Jimmy Fallon." Vedder will join him for "Comfortably Numb," which already has us drooling. Here"s the thing: the show is riddled with Vedder"s favorite artists. He could also show up with Springsteen or The Who, since he loves them both.
*Bruce Springsteen"s opening set. The performance order hasn"t been officially announced, but word has leaked out that Springsteen and the E Street Band will open. While hardcore fans are hoping for "Sandy (Fourth Of July)," it"s too obscure for a worldwide audience. Expect "My City of Ruins" and "We Take Care Of Our Own."
*Eric Clapton's guests: No word yet on who the rock god is performing with, but his dressing room says "Eric Clapton and Friends," according to one of our spies, so expect some high profile artists, who aren't already announced to possibly join him.
*Billy Joel, whose modified performance of "Miami 2017" was the hands-down highlight of the first Sandy benefit on NBC on Nov. 1 will play with his full band for the first time in three years. He will have no special guests with him.
*The surprises. There are some names that haven"t been officially announced. One rumor we"ve heard but weren"t able to confirm that makes perfect sense is Paul Simon. How could he not be there? We heard his name from one person on the scene, while another source told us his name hadn't appeared on any schedules. The other obvious one is Jay-Z. Though he"s not listed, we wouldn"t be surprised if he"s a walk-on at some point, perhaps during Alicia Keys" set.
*Paul McCartney's closing jam. There"s always one for something like this where everyone still in the building shows up for a rag-tag cover. Showbiz 411 is reporting that McCartney will close the set, so expect some kind of sing-along on either "Hey Jude" or another Beatles classic, but I wouldn"t bet against "Hey Jude."
Proceeds from the evening go to The Robin Hood Relief Fund. To donate, click here. The concert starts at 7:30 p.m. ET and will air basically everywhere, including 32 TV channels and on the internet.


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03:55PM, Friday 01 February 2019
The winners of the Estrella Damm Top 50 Gastropub Awards were announced at Lillibrooke Manor on Tuesday.
Berkshire and Buckinghamshire establishments held their own at the event, with a generous handful of pubs gaining places on the prestigious list.
The awards event, which is in its 10th year, was presented by Angellica Bell, known for The One Show and winning Celebrity MasterChef in 2017.
With two of his pubs in the top 50 list, Marlow-based celebrity chef Tom Kerridge was well placed as the person to cook the celebratory lunch on the day.
Kerridge's The Coach, in West Street, was awarded second place and The Hand and Flowers, also in West Street, was put in 14th place.
The awards are broken down to cover pubs in six geographical regions and voted for by 'top foodies and hospitality experts' including chefs, food writers and pub guide editors.
Also in the top 10 was The Beehive in White Waltham, which came in at number eight on the list. The Crown at Burchetts Green is placed in 13th, in at 26 is the Royal Oak Paley Street and number 48 is The Oxford Blue in Windsor.
Editor of the Estrella Damm Top 50 Gastropubs Nicholas Robinson said: "Gastropubs are an example of everything that is great about the UK food and drink scene.
"It is something no one else in the world can do as well as we do in Britain. As a result, the pubs on the list are envied by so many because of the innovation and skill within them.
"Real gastropubs offer unique experiences while remaining focal points within communities. They are very special places to be."
To see the list go to www.top50gastropubs.com
Editor's Picks
Most read
Top Ten Articles
At around 9.30am today (July 17) a red Honda motorcycle and a white Ford Transit van were involved in a collision on the A308 Marlow Road between Maidenhead and the Bisham roundabout.


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Updated In his second new TV spot of the day — this one now playing in Pennsylvania — Barack Obama takes a strong stand against Big Oil, saying he "won't let them block change anymore."
Senator Obama stands at a gas station and speaks directly into the camera for the duration of the 30-second spot. (Must be his new style — he takes a similar direct approach in his Indiana ad.)
Mr. Obama goes after Exxon Mobil in particular. in the ad, which starts off with old footage of cars lined up for gas at an Exxon station back in the 1970s.
People cried out for energy independence back then, Mr. Obama says, and "nothing's changed — except now Exxon's making $40 billion a year, and we're paying $3.50 for gas."
But he's going to do something about it, he says. Mr. Obama proposes putting a tax on the windfall profits of oil companies, and he says he'll end American dependence on foreign petroleum. He can take aggressive steps against Exxon Mobil, he points out, because he hasn't accepted donations from oil companies or lobbyists.
"It's time that Washington worked for you. Not them," he says.
Hillary Rodham Clinton's campaign was quick to send out an e-mail accusing Mr. Obama of making false statements in his ad, saying he has received more than $160,000 from the oil and gas companies.
Phil Singer, deputy communications director for Mrs. Clinton, put out this statement: "It's unfortunate that Senator Obama is using false advertising to explain why he can be trusted to do something about energy prices. Senator Obama says he doesn't take campaign contributions from oil companies, but the reality is that Exxon Mobil, Shell and others are among his donors. I wonder if they'll fix the ad."
Mr. Singer shouldn't hold his breath. Bill Burton, press secretary for the Obama campaign, reaffirmed the ad's message, saying "Senator Obama is the only candidate in the race who doesn't accept campaign contributions from special interests PACs and Washington lobbyists, and that includes oil companies and oil lobbyists."
"The energy bill that Senator Clinton has already been criticized for misrepresenting — one that Clinton supporters Representatives Murtha and Kanjorski also backed — actually raised taxes on oil companies and made the largest investment in renewable energy in our nation's history," said Mr. Burton in a statement. "Instead of continuing with the negative and misleading tactics that voters everywhere are rejecting, Senator Clinton should get behind the Obama plan to ease the burden of rising gas prices on working families."
Here's the script for "Nothing's Changed":
Since the gas lines of the '70's, Democrats and Republicans have talked about energy independence, but nothing's changed — except now Exxon's making $40 billion a year, and we're paying $3.50 for gas.
I'm Barack Obama. I don't take money from oil companies or Washington lobbyists, and I won't let them block change anymore. They'll pay a penalty on windfall profits. We'll invest in alternative energy, create jobs and free ourselves from foreign oil.
I approve this message because it's time that Washington worked for you. Not them.


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More About Domino's on Paradise Rd
Order Fresh Handcrafted Pizza at 430 Paradise Rd
When your tastebuds are tingling for a delicious slice of cheesy, meat-topped goodness, we know you're not craving any old pizza in Swampscott. You're looking for a flavorful pizza with a freshly made tomato sauce, expertly sliced pepperoni, heaping piles of melted mozzarella and a hand-tossed crust that's both chewy and crispy. You want an undeniably good pizza. You want Domino's.
At Domino's on Paradise Rd, we make it fast and easy to get the fresh pizza you're craving. Order your pizza in person, give us a call at (781) 715-2220, or place your pizza order online!
Get started by picking one of our homemade crusts, including a fluffy, seasoned hand-tossed crust, a pan crust featuring two huge layers of cheese, a crispy thin crust, or a gluten-free pizza crust! Next, pick your sauce and start piling on your favorite toppings. Whether you can't get enough grilled chicken and roasted veggies or you adore a savory variety of banana peppers, mushrooms, and pepperoni, you can be sure your local Domino's has it all.
With over 34 million different ways to make a single Domino's pizza, you can be sure we've got a slice for any taste. Customize your pizza from bottom to top or choose from one of our chef-inspired specialty pizzas, like the popular MeatZZa™, Honolulu Hawaiian, or Memphis BBQ Chicken, and have it delivered to your door today, fast!
Like A Butler, But For Pizza: Swampscott Pizza Delivery Near Paradise Rd
At your Domino's on Paradise Rd, we don't take pizza delivery lightly. In fact, we promise your pizza will arrive hot and fresh at your door, right out of our oven. Stay up to date with the status of your order using our custom made TrackerⓇ. Enter in your phone number online and follow the progress of your pizza order from the moment you place it until it arrives at your door... and try to keep your drool from falling onto your keyboard or phone in the meantime.
Call (781) 715-2220 or order pizza delivery online now! Why keep your hunger waiting any longer?


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Doctors at Methodist Mansfield Medical Center said they have seen three times the amount of flu cases compared to last year.
In fact between Sept. 1 and Dec. 15 of last year, the hospital had about 100 cases. This year they have about 300 cases, a 300 percent increase.
The hospital isn't just dealing with patients walking in, but some of the patients are people who work at the hospital, as well. One of those cases, Renee Trussell's 20-month-old-daughter. Trussell runs the gift shop at the hospital.
The latest news from around North Texas.
"This is the first time we've gotten the flu in our family," explained Trussell. "She did get the flu vaccine and she got Tamiflu, and she's doing a lot better today."
Case load manager Jessica Young came down with the flu last week and is still fighting it off.
"I started having terrible body aches all over and a low-grade fever," said Young.
The head of the Emergency Department called it a vaccine failure.
"The strain is not well covered by the vaccine," explained Dr. Keto Trivedi. "Not only are we seeing patients that never got the flu vaccine that are getting the flu, but people who also go the vaccine that are still getting the flu."
Trivedi said it's better to get the shot and have some form of protection, than having none at all.
As the week before Christmas winds down, Trivedi said the flu season is only ramping up.
"All the travel, all the family gatherings that will be occurring," he said. "I think it will get a little worse before it gets better."
The Mansfield Independent School District reported there has not been a dramatic decrease in attendance because of the flu.


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I need help. I trust in Jesus and he is probably doing this as part of his plan for my following in his footsteps while he develops a plan for me, but I am suffering in the meantime and though I am honored to follow the plan he fashions for me, some relief would help in the meantime if he will let me accept it.
Here are the biggest problems:
Knees (worse), but all joints are subject to the arthritis – has to be joints since it isn't the trigger points from my Fibro which Is currently about a 5/10. I have been using Arnica Cream, best out of what I have on hand. Prescription Voltaren is useless.
IBS – no matter whether I eat healthy or junk, it gives me terrible stomach pain and accompanying constipation and then quickly changes to diarrhea. What am I supposed to eat.
Acid Reflux – ugh. Wake up choking on acid and then swallow it. In misery for hours. Trying to sleep as upright as I can, but three pillows so far and useless. 2 straight days. Been in bed 2 days.


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Find Help
Eating disorder awareness that encourages positive self-esteem is part of the Task Force goal. Encouraging healthy self-awareness and the elimination of a "diet culture" is promoted by task force members. Learning about media's portrayal and purpose of curing problems they may not even have been through are also part of the education ammunition to arm everyone against the threat of an eating disorder.


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Welcome to the Website for the Hispanic Association of Colleges and Universities (HACU).
Why does HACU exist? There is a whole series of reasons but one overwhelming need, to improve higher education access and success for Hispanic students.
Hispanics, a Federal designation that includes Mexican Americans, Puerto Ricans, Cuban Americans, and other U.S. citizens and residents who trace their roots from Latin American countries and Spain, are now the largest minority group in the U.S., comprising over 17% of the population. Moreover, it is the youngest and second fastest-growing population group in the United States.
The Bureau of Labor Statistics points out that Hispanics now make up half of the increase in the American workforce. Latinos are the future of the American economy.
But Hispanics have faced a range of barriers, economic and social, to educational attainment. Historically they suffer from the highest high school dropout rate of any population group. Consequently Hispanic college-going and graduation rates lag those of others.
While traditionally Hispanics have been concentrated in the Southwestern United States, in Florida, New York and Illinois, and of course in Puerto Rico, large numbers of them are emerging in other regions and states. It is not surprising then that most Latinos attend college in those same regions.
One of the striking facts that gave rise to HACU is that less than 15% of the more than 3500 colleges and universities in the U.S. educate more than 60% of all college-going Latinos. These Hispanic-Serving Institutions (HSIs) are defined as colleges and universities whose enrollments are at least 25% Hispanic.
Another group of institutions, HACU's Associate Members, fall short of the 25% requirement, but have at least 1,000 Hispanic students enrolled or a minimum of 10% Hispanic enrollment. In more recent years, HACU has embraced international members, colleges and universities from throughout the Spanish- and Portuguese-speaking world, as well as Partner Institutions, who while they do not have the Hispanic enrollment numbers, are yet committed to improving higher educational opportunities for Hispanics. Together, HACU member institutions educate two-thirds of all U.S. Hispanics in higher education.
Adding to the challenge is the underfunding of Hispanic-Serving Institutions. In 2010-11 HSIs on average received $3,815 in federal funding per student, compared to $5,554 per student all institutions of higher education received. The disparity continues even after some years of federal funding targeted to HSIs, in large part, through the advocacy efforts of HACU.
As you browse our website, you will see the variety of activities that HACU has undertaken to address these issues. Government Relations catalogues our public policy efforts to address these issues. Programs and Student Resources, including the HACU National Internship Program (HNIP), present a variety of efforts to directly improve Hispanic educational success by reducing the barriers and increasing resources available to HSIs.
HACU is especially grateful for the sponsors and partners, federal agencies, corporations large and small, and foundations and non-profits that support us in all these efforts.
We hope that as you learn more about the critical importance of these issues, you will also join the champions of Hispanic success in higher education.
Antonio R. Flores, Ph.D.


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Founder's Corner: Gina Bianchini of Mighty Networks on goal setting, recruiting, and staying customer focused
Gina explains why enabling people to connect around their passions is important for today's society. Along the way, she also discusses:
- Her unique perspective on building a company in a competitive market
- How she approaches recruiting from a career development standpoint
- How she manages her time as a CEO
- Why she starts each day by outlining intentions and goals
Gina has a storied history building social networks and communities online. Before starting Mighty Networks, Gina cofounded Ning with Marc Andreessen and pioneered social community building. Under her leadership, the platform grew to 90 million people across 300,000 active communities in entertainment, politics, and education.
Listen to the full episode here.
Subscribe to Founder's Corner on iTunes, SoundCloud, or Overcast, and learn more about Mighty Networks here. Join the conversation with Shripriya Mahesh and Gina Bianchini on Twitter, @OmidyarNetwork #ONFoundersCorner.
Omidyar Network's Emerging Tech team invests in early-stage startups that aim to build great for-profit businesses that can change the world and achieve positive societal benefits at scale.