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X-linked adrenoleukodystrophy (X-ALD) is a fatal neurodegenerative disease caused by mutations in the ABCD1 gene, encoding a member of the peroxisomal ABC transporter family. The ABCD1 protein transports CoA-activated very long-chain fatty acids (VLCFAs) into peroxisomes for degradation via β-oxidation. In the severest form, X-ALD patients suffer from inflammatory demyelination of the brain. As the extent of the metabolic defect in the main immune cells is unknown, we explored their phenotypes concerning mRNA expression pattern of the three peroxisomal ABC transporters, VLCFA accumulation and peroxisomal β-oxidation. In controls, ABCD1 expression was high in monocytes, intermediate in B cells and low in T cells; ABCD2 expression was extremely low in monocytes, intermediate in B cells and highest in T cells; ABCD3 mRNA was equally distributed. In X-ALD patients, the expression patterns remained unaltered; accordingly, monocytes, which lack compensatory VLCFA transport by ABCD2, displayed the severest biochemical phenotype with a 6-fold accumulation of C26:0 and a striking 70% reduction in peroxisomal β-oxidation activity. In contrast, VLCFA metabolism was close to control values in B cells and T cells, supporting the hypothesis that sufficient ABCD2 is present to compensate for ABCD1 deficiency. Thus, the vulnerability of the main immune cell types is highly variable in X-ALD. Based on these results, we propose that in X-ALD the halt of inflammation after allogeneic hematopoietic stem cell transplantation relies particularly on the replacement of the monocyte lineage. Additionally, these findings support the concept that ABCD2 is a target for pharmacological induction as an alternative therapeutic strategy.
X-linked adrenoleukodystrophy (X-ALD; Phenotype MIM number #300100), the most frequent monogenetically inherited peroxisomal disease, causes demyelinating and neurodegenerative processes in the nervous system (1–3). The broad clinical presentation can be grouped into two major phenotypes, adrenomyeloneuropathy (AMN) and inflammatory cerebral adrenoleukodystrophy (CALD). AMN (mean age-of-onset 28 years) is thought to be the default manifestation of X-ALD, a slowly progressive non-inflammatory axonopathy involving the spinal cord and peripheral nerves. This results in a typical triad of spastic paraplegia, sensory involvement and bladder dysfunction (1). In CALD, rapidly progressive, inflammatory cerebral demyelination occurs independently of AMN. Being the most severe form of X-ALD, patients suffer from rapid cognitive and neurological decline and within few years proceed to a vegetative state. For all male X-ALD patients, there is a 60% risk to develop CALD. Most commonly, CALD occurs during childhood before the onset of AMN (35–40%; mean age-of-onset 7 years) and less frequently (20%) in adolescent or adult AMN patients (3,4). About 66% of male X-ALD patients have primary adrenocortical insufficiency (Addison disease). Heterozygous females often develop a milder form of AMN, but rarely adrenal insufficiency (<5%) or the devastating brain inflammation (1,3). If symptoms exceed those of AMN, other explanations than single ABCD1 mutations must be considered (5).
X-ALD is caused by mutations in the ABCD1 gene, which encodes the ABCD1 protein (formerly adrenoleukodystrophy protein, ALDP), constituting a half ATP-binding cassette (ABC)-transporter in the peroxisomal membrane (6). There is no general genotype–phenotype correlation determining the severity of the disease (2). ABCD1 transports CoA-activated, saturated very long-chain fatty acids (VLCFAs; carbon chain length ≥22 C atoms) from the cytoplasm into peroxisomes for degradation by β-oxidation (7,8). Thus, in ABCD1 deficiency, very long-chain fatty acyl-CoAs are enriched in the cytosol. They can be further elongated by enzymes of the ELOVL (elongation of VLCFAs) family and may be incorporated into different lipids such as phosphatidylcholine, gangliosides or sulphatides and lipoproteins (9). This pathognomonic accumulation of VLCFA in cells and body fluids is used as a diagnostic criterion for X-ALD (2).
The molecular mechanisms underlying the different forms of X-ALD (AMN and CALD) are fundamentally different from each other (10). Moreover, even within different phases during development of the cerebral inflammation and the ineffectiveness of anti-inflammatory therapies, different processes and cell types seem to be involved (10). The systemic circulating immune cells may be crucial for the phenotype. At an early stage of brain inflammation, allogeneic hematopoietic stem cell transplantation (HSCT) can be applied. This is currently the only curative treatment option for CALD (11). Beginning demyelination and inflammation can be detected in brain magnetic resonance imaging (MRI), preceding the first symptoms of CALD (12). In addition, decreased magnetic resonance perfusion imaging appears to be an early predictor of lesion progression in CALD (13). Therefore, MRI plays an important role in the monitoring of patients. The 'time window' for finding an appropriate donor is narrow, because the disease progresses rapidly at that stage; and it takes at least 6 months after HSCT for the inflammation to halt (14). Moreover, for some patients no compatible donor is available. In order to circumvent this limitation, Cartier and colleagues developed a protocol for autologous HSCT. They corrected the patients' own CD34+ stem cells ex vivo with a lentiviral vector encoding intact ABCD1 protein. Using this strategy, they cured the cerebral inflammation of two childhood CALD patients (15). In spite of this success, currently there is no treatment available for CALD patients with advanced inflammation at diagnosis, for AMN patients, and for symptomatic heterozygous females. This leaves the majority of X-ALD patients without effective treatment options. Thus, there is still an urgent need to develop novel therapeutic strategies for X-ALD (11).
One principle for a novel therapeutic approach is the induction of the homologous ABCD2 gene encoding ABCD2 protein (formerly adrenoleukodystrophy related protein, ALDRP), also localized in the peroxisomal membrane. Functional overlap has been demonstrated in vitro in X-ALD fibroblasts (16) and in vivo in a mouse model of X-ALD (17), where overexpression of ABCD2 could compensate for ABCD1 deficiency. Most likely, due to differing expression levels of the two genes in different cell types and tissues, endogenous ABCD2 cannot replace ABCD1 functionally (18,19).
Our objective for the present study was to determine whether the main immune cell types are equally affected in X-ALD. Therefore, we hypothesized that the expression level of ABCD2 is a critical determinant for manifestation of the pathological phenotype in the different immune cell types of X-ALD patients. Here, we show that the three peroxisomal ABCD transporters (ABCD1, ABCD2 and ABCD3) are differentially expressed in the various immune cell types. The expression profiles support the concept of ABCD2 as a crucial modulator of the severity of the defect in VLCFA degradation in immune cells in X-ALD.
ABCD1 and ABCD2 are differentially expressed in CD34+-derived immune cells
In order to explore the biological relevance of ABCD1 in the major CD34+-derived immune cell types, we compared the basal ABCD1 mRNA levels with those of the other peroxisomal ABC transporters, ABCD2 and ABCD3 in healthy male adults (25–48 years of age) by using quantitative reverse transcription (qRT)-coupled PCR. Venous blood was collected in the morning under fasting conditions. Purity of isolated cell types was typically ≥95% as determined by flow cytometry analysis (Supplementary Material, Fig. S1). The gene-specific differences between the cell types were quite distinct and consistent. In general, there was a surprisingly low inter-individual variability in the mRNA expression profile of the three genes (Fig. 1A; cf. raw data in Supplementary Material, Fig. S2). The results are depicted as a ratio to HPRT mRNA levels, which shows minor variability across different cell types (Supplementary Material, Fig. S2). Among the analysed cell types, ABCD1 mRNA levels were the highest in neutrophil granulocytes (CD15+/CD16+) followed by monocytes (CD14+). For qRT-PCR, we separated neutrophils from the minor eosinophil granulocyte fraction (CD15+/CD16−), as neutrophils are the most important granulocyte population during inflammation. However, eosinophils are transcriptionally more active than neutrophils (20). ABCD1 mRNA levels were intermediate in B cells (CD19+) and lowest in natural killer (NK) cells (CD3−/CD56+), NKT cells (CD3+/CD56+) and T cells (CD4+ and CD8+). In contrast to the relatively high mRNA levels of ABCD1 in monocytes and granulocytes, the ABCD2 mRNA was barely detectable in these cells (Fig. 1A). In B cells and NK cells, the mRNA levels of ABCD1 and ABCD2 were comparable. However, in T cells (CD4+ and CD8+), we found the highest ABCD2 mRNA levels, exceeding those of ABCD1 by an order of magnitude (Fig. 1A). The ABCD3 mRNA was rather equally distributed in all investigated immune cells, with slightly lower levels in neutrophil granulocytes and NKT cells (Fig. 1A). This is also in good agreement to previous studies showing that the ABCD3 gene is ubiquitously expressed (21).
In order to avoid influences resulting from a biased selection of a single housekeeping gene (HPRT), we investigated the ABCD 1–3 expression levels normalized to an additional housekeeping gene (GAPDH). The results depicted as relative mRNA levels based on the geometric means of HPRT and GAPDH, show a similar profile (Supplementary Material, Fig. S3).
No compensatory regulation of the three peroxisomal ABC transporters in immune cells of AMN patients
Next, we analysed the basal mRNA levels of the peroxisomal ABC transporters in the immune cell types of X-ALD patients in order to investigate potential compensatory regulation of gene expression. We recruited adult AMN patients (29–43 years of age) with a pure non-inflammatory AMN phenotype and without Lorenzo's Oil treatment. We focused on this group of patients in order to determine the intrinsic cell properties in ABCD1 deficiency, without confounding secondary alterations due to inflammatory responses. We expect the results obtained in AMN to be representative for all forms of X-ALD in males, as the extent of VLCFA accumulation does not differ between CALD and AMN patients when comparing plasma or fibroblasts (1). The same cell populations as for the healthy controls were isolated from five AMN patients, which all had normal white blood cell counts at the time of analysis (Supplementary Material, Table S1). The mRNA levels of ABCD1, ABCD2, and ABCD3 in the patient collective (Fig. 1B) showed a similar distribution as in healthy controls (Fig. 1A). None of the AMN patients carried a mutation affecting the transcription of the ABCD1 gene. There was no statistically significant compensatory induction of ABCD1, ABCD2 and ABCD3 in any cell type of AMN patients (Fig. 2). Taking into account that upon overexpression ABCD2 can compensate for ABCD1 deficiency, these results imply that in AMN monocytes and granulocytes should be most severely affected.
The extent of VLCFA accumulation in AMN differs widely between the immune cell types
All tissues, body fluids and cell types that have been investigated from X-ALD patients accumulate saturated VLCFA, albeit to variable degrees (1). Therefore, we asked to what extent the different immune cell types isolated from blood of AMN patients accumulate VLCFA. We measured the total amount of the fatty acids (FAs) C26:0, C24:0, C22:0 and, for normalization, C16:0 by using gas chromatography–mass spectrometry (GC–MS) in the following isolated cell types: monocytes (CD14+), granulocytes (CD15+), B cells (CD19+), NK cells (CD3−/CD56+) and T cells (CD3+) of controls and AMN patients (Fig. 3). The relative amount of each VLCFA is displayed as a ratio to C16:0, a long-chain FA that does not differ between healthy controls and AMN patients (Fig. 3D; Supplementary Material, Fig. S5D). In healthy controls the absolute amounts of C26:0 were approximately 1/20-1/40 of C24:0 and 1/20 of C22:0 (Supplementary Material, Fig. S4 and S5). Among the cell populations isolated from healthy controls, granulocytes (CD15+) and B cells (CD19+) displayed the highest levels of C26:0, whereas monocytes, NK cells and T cells contained about half as much (Fig. 3A, left panel; Supplementary Material, Fig. S5A, left panel). Monocytes and granulocytes showed higher levels of C24:0 than NK cells, B cells and T cells (Fig. 3B, left panel; Supplementary Material, Fig. S5B, left panel).
When comparing the cell type-specific levels of C26:0/C16:0 of the AMN patients and healthy controls, we found a statistically significant 6-fold increase in monocytes and a 3-fold increase in granulocytes and NK cells. In contrast, B cells and T cells showed 2-fold accumulation of C26:0 compared with the control group (Fig. 3A, right panel and 3D). Similar results were obtained when considering the absolute amounts (Supplementary Material, Fig. S5A) or the ratio C26:0/C22:0 (Supplementary Material, Fig. S5E). Thus, cell types displaying high levels of ABCD1 mRNA and low or no detectable expression of ABCD2 showed the highest degree of C26:0 accumulation in AMN patients.
We observed no statistically significant differences in the C24:0 levels between AMN patients and healthy controls (Fig. 3B and D). However, in all cell types of the AMN group there was a trend towards decreased C22:0 levels, which was most pronounced in monocytes (P-value 0.08) (Fig. 3C and D).
For diagnostic purposes in X-ALD patients frequently leukocytes are used. Diagnostic leukocyte preparations contain ∼50–70% granulocytes, 3–10% monocytes and 20–25% lymphocytes. As the cell types with the highest C26:0 levels are the most abundant in such preparations, the total C26:0 accumulation was reliably established (at least 5-fold) in our patient collective (data not shown) and (22).
β-Oxidation activity reflects the extent of VLCFA accumulation in immune cells from AMN patients
The accumulated VLCFA can be derived from external sources or from endogenous FA chain elongation. Thus, the accumulation does not necessarily reflect the degradation of VLCFA. Therefore, we investigated the rate of peroxisomal β-oxidation. Because of the limited biological material and the relatively high amounts needed, the β-oxidation measurements were restricted to the most abundant immune cells, namely monocytes, B cells and T cells.
In healthy controls, the peroxisomal (substrate C26:0) and mitochondrial (substrate C16:0) β-oxidation activity of monocytes was ∼3- to 4-fold higher than those rates of B and T cells (Fig. 4; cf. raw data Supplementary Material, Fig. S6). The metabolism of C16:0 was not affected in any of the analysed cell types of AMN patients (Fig. 4B). However, we found a highly significant decrease of 69.7% in the C26:0 β-oxidation activity of AMN monocytes compared with the healthy controls. In contrast, in AMN T cells, there was a trend toward a decrease in the C26:0 β-oxidation activity that did not reach statistical significance. In AMN B cells, the β-oxidation activity was approaching control values (Fig. 4A).
Taken together, these results provide further evidence that in X-ALD, VLCFA metabolism is differentially affected in the various immune cell types, ranging from severe in monocytes to moderate in T cells and no impairment in B cells.
To date, transplantation of allogeneic hematopoietic stem cells or autologous hematopoietic CD34+ stem cells corrected by gene therapy have been the only effective treatments for CALD (14). However, the mechanisms behind the benefit of these therapies are poorly understood. Here, we attempt to shed light on the pathophysiology of the CD34+-derived cell types, as one or more of these must play a crucial role in HSCT. In addition, we identified potential target cells for future therapeutic strategies, such as pharmacological upregulation of the redundant ABCD2 gene to ameliorate the metabolic impairment and possibly halt the inflammation in CALD.
For the first time, the expression profiles of the peroxisomal ABC transporters and the biochemical phenotype were directly compared in isolated CD34+-derived immune cell types of AMN patients and healthy controls. We found that the severity of the impairment in VLCFA metabolism differs strongly between these cell types and correlates with the expression levels of the peroxisomal ABC transporters ABCD1 and ABCD2.
We identified two cell types with a robust pathognomonic phenotype, monocytes and granulocytes, which in healthy controls have a high expression of ABCD1 and no or barely detectable expression of ABCD2 mRNA in common. In these cells, we observed the highest accumulation of VLCFA in X-ALD, and in monocytes peroxisomal β-oxidation activity for C26:0 was significantly reduced (<30% of control values). Interestingly, these two cell types are functionally related and also build a cluster on the basis of gene expression (23). Our findings that monocytes are severely affected by the disease are in line with several other publications. Increased cytokine levels, especially tumor necrosis factor (TNF)-α, were previously detected in peripheral blood mononuclear cells (PBMCs) from X-ALD patients stimulated by lipopolysaccharide (LPS) (24); additionally, elevated levels of interleukin (IL)-2 and IL-1α were found (25). Most likely, this TNF-α production came from peripheral monocytes since stimulation by phytohaemagglutinin, a T cell activator, did not increase TNF-α amounts (24). Also in a mouse model of X-ALD, Yanagiasawa and coworkers showed that peritoneal macrophages from Abcd1-deficient mice secreted significantly elevated levels of TNF-α, IL-6 and IL-12p70 upon stimulation by LPS and interferon-γ (IFN-γ) compared with wild-type mice (26). Chitotriosidase activity, an activation marker for macrophages, was found to be elevated in plasma and cerebral spinal fluid (CSF) of CALD patients (27). Recent findings also showed elevated proinflammatory chemokines (IL-8, IL-1ra, monocyte chemotactic protein (MCP)-1, macrophage inflammatory protein (MIP)-1β) that induce migration of peripheral immune cells to the site of inflammation in CSF of CALD patients. Interestingly, IL-8 and MCP-1 levels correlated significantly with MRI severity, whereby IL-8 is speculated to mediate blood–brain barrier breakdown, and MCP-1 is a chemotaxin for monocytes (28). Taken together, these observations not only establish a major role for monocytes in the pathology of CALD but also provide evidence for functional abnormalities.
Here, we found that monocytes have a severe intrinsic metabolic defect in X-ALD. We suggest that this is responsible for an altered cytokine production leading to further changes in the immune response, for example T cell activation in a predominant Th1 response (25). Our results show that T cells of X-ALD patients express high levels of ABCD2 mRNA and hardly accumulate C26:0 (<2-fold increase) compared with controls. Thus, in X-ALD, primary T cells, in contrast to monocytes, apparently have only a subtle defect in their VLCFA metabolism.
Strikingly, B cells, which express intermediate levels of ABCD2 mRNA, are barely affected metabolically by ABCD1 deficiency; VLCFA do not accumulate and the β-oxidation activity for C26:0 is similar to that of controls. These results differ from published reports of 3- to 7-fold accumulation of C26:0 in a human X-ALD B-cell line (29,30). However, there are major differences in experimental settings that probably accounts for this discrepancy. Whereas we analysed freshly isolated primary B cells without any further handling or cultivation steps, Uto and colleagues (29) used lymphoblast cell lines in long-term cultures containing 10% fetal bovine serum, which has been shown to alter the expression levels of the peroxisomal ABC transporters (31,32).
Taken together, we observed that in cells with minimal or no compensatory ABCD2 (monocytes and granulocytes), the VLCFA accumulation was the most severe (3- to 6-fold) in X-ALD. However, when ABCD2 is expressed at moderate levels, as for example in B cells and NK cells, or at high levels, even exceeding that of ABCD3 mRNA, as in T cells, the extent of accumulation of C26:0 in X-ALD cases was <2-fold. Thus, in ABCD1 deficiency there is an inverse relationship between the level of ABCD2 and the extent of the metabolic impairment.
The trigger(s) for the cerebral inflammation in X-ALD are not known. Spontaneous myelin breakdown, due to membrane destabilization caused by the incorporation of abnormal VLCFA into myelin-specific lipids has been suggested as a cause for the initiation of inflammation (33). Alternatively, traumatic brain injury could act as an extrinsic factor, promoting an opening of the blood–brain barrier initiating inflammation and demyelination (34). In both cases, it is plausible that glycolipid-restricted T cells recognize their antigen within the brain. Probably the antigen is presented by CD1 molecules on perivascular cells in an MHC class I independent manner leading to a further recruitment of proinflammatory cells (35). As shown by Ito and co-workers, CD8+ T cells invade the brain at an early stage of inflammation and cause cytolysis of oligodendrocytes (35). CD8+ T cells seem to play a crucial role in the initiation and progression of the inflammation, but as shown in the present study, have no severe intrinsic metabolic defect. Thus, in X-ALD, T cells are able to execute their immune functions. However, their response might be altered due to changed cytokine production by macrophages.
Monocytes, the progenitor cells of macrophages, and granulocytes are key players in inflammatory processes. Further differentiation of monocytes into macrophages with recombinant human macrophage colony-stimulating factor did not change the ABCD2 expression status (remained below the level defined as being expressed) (36). Thus, we postulate that in CALD, the invading activated macrophages have the same metabolic defect as their CD14+ precursor monocytes investigated in our study. In order to handle the increasing FA uptake from phagocytosed myelin debris - in the absence of appropriate capacity for degradation of VLCFA - these are esterified to cholesterol, as observed by lipid analysis of autopsy material from affected lesions (37). Intracellular VLCFA-cholesterol esters deposited inside macrophages have been visualized as crystalline structures (38) and are expected to promote cellular stress. It has been demonstrated that the immunoreactivity of oxidative stress markers correlates with the degree of inflammation and myelin breakdown (39). Moreover, Eichler and co-workers showed that VLCFA-enriched lipids, in particular the lysophosphatidylcholine fraction, can lead to microglia activation and apoptosis (40). Thus, next to brain derived cell types the intrinsic molecular defect in circulating monocytes/macrophages seems to be involved in progression of inflammation.
The metabolic defect in macrophages may also provide an explanation for the failure of anti-inflammatory treatment attempts for CALD, as they do not target the intrinsic defect of macrophages. Thus, it is tempting to speculate that correction of the transport of VLCFA for peroxisomal β-oxidation in invading macrophages could halt brain inflammation in CALD. The lack of generalized genotype–phenotype correlation in X-ALD is obvious from the fact that the same familial mutations in ABCD1 can give rise to all different phenotypes (10). No mutations have been found in the ABCD2 gene of X-ALD patients and genetic association studies indicated that ABCD2 is unlikely to be a genetic modifier locus (41). However, as ABCD2 is barely expressed in some cell types including the monocytic lineage, the level of expression in particular cell types could still be a crucial determinant of the phenotype. Therefore, one possible way to restore the metabolic status of macrophages could be mediated by an induction of ABCD2 in this cell type (11). A previous clinical trial investigated the ability of the histone deacetylase inhibitor valproic acid to normalize VLCFA accumulation in X-ALD patients via an up-regulation of ABCD2 (42). However, the results were inconclusive because PBMCs were analysed and they represent a mixed cell population of mostly T cells (60–75%) and only 10–30% monocytes.
In conclusion, we provide evidence that it is crucial to analyse distinct cell types to evaluate the success of treatment at the level of gene expression or biochemical parameters. Our results show that monocytes rather than lymphocytes are the most severely affected immune cell population that needs to be targeted by therapeutic intervention in X-ALD. Furthermore, our findings provide a molecular basis for explaining why allogeneic and autologous HSCT are effective at arresting the cerebral inflammation, namely by the replacement and the functional metabolic restoration of the monocyte lineage.
Patients and healthy volunteers
In the study included were five AMN patients and 10 healthy volunteers, all male and of Caucasian origin (age 25–48 years). Patients with 'pure' AMN had isolated spinal cord disease with or without adrenal insufficiency but without cerebral demyelinating lesions at brain MRI. White blood cell counts did not reveal any signs of inflammation (Supplementary Material, Table S1). A total volume of 300 ml blood was collected with informed consent in accordance to the Declaration of Helsinki including all actual revisions. The study was approved by the Ethical Committee of the Medical University of Vienna.
Pre-enrichment of PBMCs and granulocytes
Blood was collected by venipuncture under fasting conditions into heparin tubes and diluted 1:1 with phosphate-buffered saline (PBS) at room temperature. To separate the PBMCs from other blood components, the diluted blood was overlaid onto tubes containing Pancoll separating medium (density of 1.077 g/ml; PAN-Biotech, Aidenbach, Germany) and centrifuged for 25 min at 500g at room temperature without brake. For granulocyte isolation, a Percoll (density of 1.131 g/ml; Sigma-Aldrich, St. Louis, MO, USA) step gradient was used with 72% (v/v) Percoll in PBS at the bottom and 63% (v/v) Percoll in PBS on top. Diluted blood was overlaid and centrifuged for 25 min at 500g at room temperature without brake. Afterwards, the PBMCs and granulocyte fraction (mainly neutrophils and ∼5–10% eosinophils) were removed and washed with PBS.
Magnetic activated cell sorting
The following microbead-coupled antibodies were used for isolation of specific cell types from PBMCs: CD3+, CD4+, CD8+, CD14+, CD19+, CD56+ and an NKT cell separation kit (Miltenyi Biotec, Bergisch Gladbach, Germany; Supplementary Material, Table S2). For the measurement of the ABC transporter mRNAs in neutrophils, a CD16+ microbead-coupled antibody (Miltenyi Biotec) was used to further separate them from eosinphils in the granulocyte fraction. Specific cell types were separated by either positive or negative selection according to the instructions of the manufacturer. In some cases, the unlabelled cell fraction (flow-through after positive selection) was recovered and used for the isolation of an additional cell type. Purity of the cells was measured on a BD FACS Calibur flow cytometer (Becton Dickinson, NJ, USA) and analysed using FlowJo software (Treestar Inc., Ashland, OR, USA).
Quantitative real-time PCR analysis
Purified cell types were lysed in Trizol following the manufacturer's (Invitrogen, Paisley, UK) instructions using 1 ml Trizol reagent per 106 cells. For homogenization, the lysate was passed through a QIAshredder spin column (Qiagen, Bothell, WA, USA). The isolated total RNA fraction was purified further using RNeasy Mini Kit and RNase-free DNase digestion (Qiagen). RNA concentration was measured based on optical density using a Nanodrop spectrophotometer (Peqlab, Erlangen, Germany). Total RNA (80 ng) was reverse transcribed using the iScript™ cDNA Synthesis Kit (Bio-Rad, Hercules, CA, USA). The cDNA was diluted 1:5 in RNase-free water and aliquots corresponding to 4 ng of total RNA were amplified in technical duplicates by qRT-PCR. ABCD1, ABCD3 and HPRT were detected by the SYBR Green method using SsoFast™ EvaGreen Mix (Bio-Rad). ABCD2 was detected by the Taqman method using SsoFast™ Probes Supermix (Bio-Rad). Both methods were carried out using the CFX96 Realtime System (Bio-Rad) according to the manufacturer's instructions. Thermocycler settings are available as supplementary information (Supplementary Material, Table S3).
For amplification and detection of ABCD1 cDNA, the primers 5′-GAGAACATCCCCATCGTC-3′ (forward, nucleotide position 1828) and 5′-TGTAGAGCACACCACCGTA-3′ (reverse, nucleotide position 1996) were used (GenBank™ Accession No. NM_000033.3). For ABCD2 cDNA, the primers 5′-TCCTACACAATGTCCATCTCT-3′ (forward, nucleotide position 1883) and 5′-AGGACATCTTTCCAGTCCA-3′ (reverse, nucleotide position 1961) as well as the TaqMan fluorescent probe 5′-Cy5-CAAAGAGAAGGAGGATGGGATGC-BHQ2-3′ (nucleotide position 1915) were used (GenBank™ Accession No. AJ000327.1). For ABCD3, the primers 5′-CGGCTCATCACAAACAGTGA-3′ (forward, nucleotide position 811) and 5′-AGGTGTTCCACCAGTTTTCG-3′ (reverse, nucleotide position 908) were used (GenBank™ Accession No. M81182.1). For HPRT, the primers 5′-CCCTGGCGTCGTGATTAGT-3′ (forward, nucleotide position 182) and 5′-CAGGTCAGCAAAGAATTTATAGCC-3′ (reverse, nucleotide position 401) were used (GenBank™ Accession No. NM_000194). For GAPDH, the primers 5′-AGGTCATCCATGACAACTT-3′ (forward, nucleotide position 560) and 5′-AGTCTTCTGGGTGGCAGT-3′ (reverse, nucleotide position 636) as well as the TaqMan fluorescent probe 5′-FAM-CATGACCACAGTCCATGCCA-TAMRA-3′ (nucleotide position 597) were used (GenBank™ Accession No. NM_002046)._Standard curves of known copy numbers were generated for quantification using linearized plasmids containing human ABCD1, ABCD2, ABCD3 and for normalization HPRT and GAPDH cDNA.
Extraction of FAs and gas chromatography–mass spectrometry analysis
The following FAs were used as standards for quantification: [3,3,5,5-2H4]-hexacosanoic acid (2H4-C26:0), [3,3,5,5-2H4]-docosanoic acid (2H4-C22:0) and [3,3,5,5-2H4]-tetracosanoic acid (2H4-C24:0) were obtained from Dr Herman J. ten Brink from the Free University Hospital, Amsterdam, the Netherlands. [7,7,8,8-2H4]-Palmitic acid (2H4-C16:0) was from Cambridge Isotope Laboratories Inc., Andover, MA, USA. Cell pellets were suspended in distilled water and sonicated three times for 30 s on ice. Protein concentration was determined by the method of Lowry using bovine serum albumin as standard (42). Extraction and GC–MS analysis was performed as described (22), using 100 µl internal standard mixture (containing deuterium-labelled free FAs including 0.1 µg/ml 2H4-C26:0, 0.5 µg/ml 2H4-C22:0 and 2H4-C24:0, respectively, and 10 µg/ml 2H4-C16:0 in toluene) per vial. Data were analysed utilizing calibration curves obtained from the appropriate non-labeled FAs.
β-Oxidation of 1-14C-labeled FAs
[1-14C]-palmitic acid (C16:0; ARC 0172A) and [1-14C]-hexacosanoic acid (C26:0; ARC 1253) were obtained from American Radiolabeled Chemicals (St. Louis, MO, USA). Free FAs in ethanol were aliquoted into glass reaction tubes, dried under a stream of nitrogen and solubilized in 10 mg/ml α-cyclodextrin by ultrasonication. β-Oxidation of labeled FAs to acetate was carried out essentially as described previously (8,43). In short, the reaction mix of 250 µl contained 4 µm of labeledFA, 2 mg/ml α-cyclodextrin, 30 mm KCl, 8.5 mm ATP, 8.5 mm MgCl2, 1 mm NAD+, 0.17 mm FAD, 2.5 mml-carnitine, 0.16 mm CoA, 0.5 mm malate, 0.2 mm EDTA, 1 mm DTT, 250 mm sucrose and 20 mm Tris, pH 8.0. Reactions were started by addition of 1–2 × 106 cells, which had been stored at −80 °C, carried out for 1 h at 37°C and stopped by addition of KOH and heating to 60°C for 1 h. After protein precipitation by HClO4, a Folch partition was carried out and 14C-acetate was determined in the aqueous phase by scintillation counting.
Statistical analyses
Linear mixed effect analyses (44–48) were used to describe the relationships between the experimental responses (mRNA copy number, FA amount and β-oxidation activity) and three primary covariates (type of gene, FA, β-oxidation) in the main immune cell types (cellType) of healthy controls and AMN patients. Replicated recordings were obtained. We modeled the subject factor as a random effect and the gene/FA/β-oxidation factor, the population (healthy controls; AMN patients) factor and the cell type factor as fixed effects. Separate fixed effect models incorporating an interaction term between primary covariate and the population factor were fitted within the levels of cell type factor. A variance stabilizing log transformation of the response was used for all analyses.
The interaction term (primary:population) enables different changes (slopes) between the two populations for each level in the primary covariates to be incorporated in the fixed effect model. All terms of the equation are defined in italics in the text.
We tested the significance of terms in the fixed effect model using the conditional F-test. A significant fixed effect term was further tested using Student's t-test. Multiple testing was taken into account using simultaneously calculated z-values for the covariates of interest, obtaining the P-values from a multivariate Student's t-distribution.
Supplementary Material is available at HMG online.
This work was supported by the EU project 'LEUKOTREAT' (241622), the Austrian Science Fund (FWF) (P26112-B19) and the European Leukodystrophy Association (ELA) Germany. Funding to pay the Open Access publication charges for this article was provided by the Austrian Science Fund (FWF) (P26112–B19).
We thank the voluntary donors, in particular the AMN patients. We thank Manuela Haberl, Eva Steinhuber and Regina Sundt for excellent technical assistance and Prof. Hans Lassmann for critical reading of the manuscript.
Conflict of Interest statement. None declared.


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Sam enjoys a vulnerable moment with Jason.
Sam spies Jason and Elizabeth sharing a tender moment.
Spinelli and Jason get to bond like the friends they used to be, with a slightly different dynamic.
Nikolas wields control over Jason.
Nikolas purposely provokes Jason into a throwdown.
Nikolas isn't just going to lie down for Jason like a bitch.
Sonny and Carly will bail Jason out of this jam.
Nik and Jason aren't the only ones locked in a battle. Watch out for Maxie vs. Valerie.
Elizabeth must have that talk with her boys, as a consequence of her fraudulent behavior.
Little Jake turns into a little demon when he sees Jason and Sam cozying up.
At some point, Jason is gonna have to talk to both Sam and Elizabeth.
Sam and Sonny find themselves remembering a fond moment from the past.
Brothers Michael, Dante, and Morgan have a chance to commiserate, with their respective sob stories.
Kiki finds she's no match for Ava's emotional manipulations.
Julian and Alexis play house with his and Olivia's baby.
Paul's evildoings will soon be unveiled for all to judge and jury.
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General Hospital 12-22-15 Full Episode
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June 13th, 2016
Episode 538 of 753 episodes
A U.K. exit from the European Union could tip the struggling region into recession, says Wharton's Mauro Guillen.
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a (possible) story: individual differences, learning, & education Or, TEP115-inspired research hypotheses
Achievement gaps: a starting point No child left behind: black/white, rich/poor gap Want all students scoring proficiency by 2014 What factors contribute to how a child is doing at school? What does this tell us about the way we learn ; about how our minds work ? Also.. The role of the school– is it the only thing? What does this mean for us as teachers?
Individual differences thoughts How does ch. 12 approach individual differences? Gender diffs, Ethnic diffs : are these the best ways to approach individual differences? Are there *really* differences between two populations? Are our chosen populations (e.g. "ethnicity") really (relevantly?) (causally?) related to the differences we see? Cognitive deficit views Contextual views Cultural "how could how much money a family has, or the colour of their skin influence how well that child learns to read?" Social class vs. income & race
Rothstein Class and schools: using social, economic, and educational reform to close the black-white achievement gap (2004). Some SES-related factors Personality & Childrearing** Health care Housing (in)stability Cultural factors Values ; parent participation Historical biases: culture of underachievement My interest: child-care factors which might lead to more or less confident/exploratory children, and how this relates to school performance Which of these factors does ch. 12 bring up?
A family of concepts Secure/anxious attachment relationships Who's learned about these before ? Exploration Pro-activity Confidence (?) Do you see any relationship between these? How about, a possible connection to learning, to success in school?
Bowlby: attachment Focused on "long term affects of different patterns of attachment of infants and young children to their caregivers" How does the child approach its local environment? Is it interested? Is it frightened? Is it approaching or retreating? Secure vs. anxious attachment Supportive responsiveness of caregiver Again, what could this have to do with learning? Does attachment relate to school performance… WHY? Exploration Pro-activity Confidence (?)
Individual differences in EA & individual differences at school: links Predictive value of early attachment ratings "Strange situation" test : measuring EA Arend, Gove & Sroufe (1979): in/secure attachment predicts how yr olds will tackle (detail) (another slide? ) Predicts school performance (controlling for IQ) Predicts curiosity and exploratory-ness as rated by teachers
Why do we see these correlations? What are possible links? What is going on? "[links] indirect": "Having a secure attachment relationship.. likely does not change your "math brain" " (Sroufe et. al, 2005, p. 187) Sroufe et al. "the central outcomes.. Are a basic sense of social connection, positive expectations concerning self and others, and the capacity for self-regulation" (p.164) Can we make any stronger hypotheses?
Stronger hypotheses The role of curiosity, exploration, pro- activity in doing well in school : pro-active learning Maybe this is the relevant link between secure early attachment and later success
Aside: But what could curiosity, exploration & pro-activity have to do with school performance ?? Japanese-US differences in teaching Japanese kids have higher scores: before they learn about a concept, teachers want kids to struggle through problems Schwartz & Moore Kids score highest in "compare -> lecture" conditions What are the similarities in these approaches? What does this have to do with (something like) pro-active learning ?
Pro-activity & school : hypotheses In situations where students are active/engaged with material: they perform better when tested. So, exploratory children, ones who are interested in alternative-testing; kids who, try out different ways of looking at a problem kids who "explore"-- do they have a better chance of general school-success? (a hypothesis) Maybe this kind of confidence is a factor in why certain students are not doing well in school.
Why are we talking about this again? Data show that SES is the highest predictor for early attachment ratings. Second highest is.. So, this is an individual difference that SES plays a role in constructing.
Other (non-attachment) factors affecting individual pro-activity Rothstein: how parents talk to their kids Low SES parents: "we're getting off here" High SES parents: embed directives in questions: "oh, look, is this our stop? " Again, how might this be related to what we've just said about the importance of pro-activity?
Directives vs. open-ended instructions Ball-game study (find reference) Part one: Two groups of college students, both taught how to play a novel game First group was given instructions as follows: "This is how you throw the ball" Second group heard "This is one way you might throw the ball" Part two: Change rules a little bit – what happens? Second group much better at playing the second game
Extensions What about things like, learning language (growing vocabulary)? Learning how to read?? We see SES differences here, quite early Are there any possible links to pro- active learning in these kind of learning tasks?
Summing up The role of the school– is it the only thing? What do effects of SES mean? Rothstein: we need social, economic, & educational reform to close the gap What does this mean for us as teachers? Involvement in reforms ! Awareness of the issues ! Teaching for exploration
Some books I'm drawing from Rothstein, R. (2004). Class and schools: using social, economic, and educational reform to close the black-white acheivement gap. Economic policy institute. Bowlby, J. (1988). A secure base: parent- child attachment and healthy human development. Basic Books. Sroufe et al. (2005). The development of the person: The Minnesota story of risk and adaptation from birth to adulthood. Guilford Press.
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Arnold Schwarzenegger, clad in a jacket and open-necked shirt, strolled onto the set of The Tonight Show With Jay Leno Wednesday to mug his way through the bombshell announcement that, yes, he was running for California governor in the total-recall election. In an Austrian accent that only Henry Kissinger could envy, the muscular movie star added to the dignity of the occasion by solemnly declaring, "It's the most difficult decision I've ever made in my entire life, except for the one in 1978 when I decided to get a bikini wax."
So it was this week with the Terminator and the Terminated, the actor who played a robot and the politician who was compared to one. While Arnold reprised his signature "hasta la vista, baby" line, Al joked about his personal role in the Bush economic downturn: "I was the first one laid off. You never forget a thing like that."
The bicoastal juxtaposition of the action hero's entrance into California's demolition-derby gubernatorial politics and the former vice president's reappearance on the political stage inevitably prompts thoughts about the use and abuse of celebrity. The calculus is more complex than simply opining that Schwarzenegger is comically ill-suited to preside over California's fiscal crisis and that Gore's emergence in the role of Democratic elder statesman is welcome.
There is a time-warp quality to wails about the danger of Hollywood hijacking politics. Back in the 1960s, not only did Ronald Reagan, the star of that celluloid classic Bedtime for Bonzo, become California's governor, but the state also elected movie hoofer George Murphy to the Senate. Comic troubadour Tom Lehrer even sang a ditty about Murphy that included the immortal line, "At last we've got a senator who can sing and dance."
But despite Reagan's two successful terms in the White House, somehow this dire trend never escalated into President Warren Beatty or Senator Charlton Heston or even Governor Eminem. The residents of Minnesota even survived four years with Jesse Ventura, a shameless wrestler who favored feathered boas in the ring, in the governor's mansion.
It now seems ludicrous, in hindsight, that the imperiled Gray Davis was first elected governor of California in 1998 with the slogan "Experience that money can't buy." As he has demonstrated, a lifetime of experience with political fundraising, backroom negotiations and in-your-face campaign tactics does not translate into stirring leadership. Although Davis may be an extreme example, career politicians are often afflicted with a truncated sense of the possible. Voters sense this, which partially explains the allure that now surrounds Dr. Howard Dean, the Marcus Welby of the Democratic Party.
So, in theory, there is no more reason to bar a movie star from being governor than there is to ban a billionaire from serving as New York City's mayor. Both Reagan and Michael Bloomberg understood the guiding principle that governs midlife career shifts into elective office: Voters demand dignity from their leaders. Schwarzenegger, judging from his appearance with Jay Leno, appears to lack that internal governor. The bikini-wax joke would have been fine for an actor joshing with a late-night TV host. But even Bill Clinton didn't reveal his preference for boxers over briefs in his speech announcing his candidacy for president.
The point is not to exalt prudishness. We live in permissive times, and we should not expect would-be political leaders to suddenly don top hats and frock coats. But for all the ways in which California politics is fast turning into a demonstration of chaos theory, the governorship of the nation's largest state is not a job for cut-ups and class clowns. A credible Schwarzenegger candidacy requires more than tough-guy poses and denunciations of the political calling.
Gore, in his speech Thursday, faced a different kind of credibility issue. It was the difficult challenge of how to appear relevant without unduly intruding into the Democratic presidential race. The former vice president made the case that the premises that governed the administration's march to war in Iraq were "just dead wrong." But Gore's meticulous argument was similar to the claims that such presidential candidates as Dean, Bob Graham and John Kerry are making on the campaign trail.
Even though Gore at one point called the ouster of Saddam Hussein "a positive accomplishment" for the president, most of his speech dripped with scorn for the administration's unilateral approach to war and the reconstruction of Iraq. His skeptical tone seemed at odds with the outspoken position of Joe Lieberman, his 2000 running mate, who just this week denounced Democrats "who said 'no' to eliminating Saddam Hussein or were ambivalent about it, before and after the war." Yet Gore never mentioned Lieberman or even directly alluded to the fissures among the Democratic presidential contenders over Iraq.
That is the choice every party elder faces — whether to wade into the midst of the controversy of the moment or to deliver well-crafted speeches devoid of much surprise or news content. It all comes back to the question of dignity. Gore these days has too much and Schwarzenegger too little.
Contributing: Walter Shapiro's column appears Wednesday and Friday. E-mail him at firstname.lastname@example.com


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Results 1 to 5 of 88
Thread: Initial Word Game
July 6th, 2016 03:07 PM #1Senior Member
- Join Date
- Jun 2016
- Posts
- 267
Initial Word Game
Alright, so this game is pretty simple. I will start off by giving you a word, for example, S E A, and the next person has to write a name with those initials, then they will give another word to the next person. Since there are going to be some words that are too long to maybe write as one entire name, I figured you could split the name up into siblings. Let's say you are given the word H A I R you could then write Holly Adelaide + India Rosamund or even split it up into individual siblings such has Harriet + Alexander + Isaac + Rebecca or a child with three names and a siblings with one, whatver you want really, just play around with it.
Here is another example:
Person 1: B I R D
Person 2: Bianca Isabel "Bambie" + Ralph Daniel --- S E P T E M B E R
Person 3: Susan Eleanor Patience + Timothy Edward Miles + Bridget Emily Rose --- S H I R T
You get the idea. So I'll start off:
C A K Evegan. teenberry. hungary-america. building the list.
hungarian lady loves:
villő, bernadett, luca
July 6th, 2016 03:16 PM #3Matilda Annabelle * Natalie Ruth * Emily Rose * Calliope Justine
Jensen Andrew * Brandon Poole * Jasper Coyote * James Blake
July 6th, 2016 04:20 PM #5Senior Member
- Join Date
- Jun 2016
- Posts
- 267
July 6th, 2016 04:22 PM #7Norwegian Berry
Clara Luella • Esther Cecilia • Agnes Matilda
Gustav Martinus • Vincent Johan • Wilhelm Edward
Alvdis • Aria • Cadence • Dagny • Eilish • Eir • Ffion • Greta • Hedda • Io • Maisie • Margo • River
Angus • Beau • Birch • Henry • Imre • Johan • Orion • Pil • Silas • Storm • Timothy • Tindre
July 6th, 2016 04:31 PM #9Teenberry~17~Ravenclaw~Whovian~INTP~Taurus
I like WAY too many names to list, but here are some of them:
Kennedy Emerson Eliot Sasha Jaxon Braxton Jase Alva Fox Asa Carbry Zia
Anderson Dara
Nova Briar Michaela Montserrat Cadence Harriet Phoenix Logan Star Justine Faith
If you would like to vote again, here's my list: http://www.behindthename.com/pnl/159828 (Last Update: January 17th, 2017)


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Would this be a good cold day activity?
Is this an outdoor attraction or activity?
Is this attraction pet friendly?
Would this be a good sunny day activity?
Would this be a good hot day activity?
Is this attraction good for couples?
Is this attraction suitable for adults only?
Is this attraction suitable for all ages?
Does this attraction require above average amounts of physical activity (long walks, climbs, stairs or hikes)?
Does this attraction have good scenic or skyline views?
Thanks for helping!
The only public lighthouse in South Carolina.
Stunning view and relatively short hike up to the top. Just beware the wind gusts - my husband offered a pair of sunglasses to the ground below!
Due to hurricane damage in October the park is currently closed (lighthouse, office, camping areas). Nature Centre is open, pier partially open. Beautiful area, still a nice drive if you are in area. Lots of birds in parking area and views from pier. One stop open on way to park but marsh boardwalk also out of commission. Park ranger told...
Love the view from the top. Fabulous over the tree tops. The climb was worth the size strategy effort for the opportunity to look out on the ocean.
This is a very nice are and is a nice ride for a little trip in the Low Country, saying that however like the rest of the east coast of the Low Country it has a lot of tree damage so the all entrances to the light house and the camping area is closed. The nature center further down the...
Go all the way to the top and take pictures of a great view. Unique museum located nearby with many historical items from bygone days of lighthouse keepers.
Lots of parking and walking areas.
If you want to see a lighthouse this is a beautiful area. They charge a couple dollars to let you walk up in the lighthouse and are really sticklers about it.
Located on Hunting Island, SC you can climb to the top and see forever! Has some old out buildings with historic references to read on the ground where the old keeper's housing was.
We actually did not go in lighthouse but walked by to the beach. Visiting family. They asked if we sished to go in lighthouse but we had little one traveling with us. Beach was nice, unique. warm shallow water. good for pics
Lots of downed trees in the area...was told the park would be closed until end of year! Do stop at Beaufort visitors center for updated information on hurricane damage...most everything else in area is open.
You can drive by the park
ALOT OF Spiral steps....minimum fee charged..beautiful view from top
Over looking ocean.
Sits on beautiful grounds
Nice state park.....Huntington island state park
Own or manage this property? Claim your listing for free to respond to reviews, update your profile and much more.
Does anyone know if the lighthouse is open? No one answers when we call and it doesn't say anything on the website for the lighthouse. Anyone been recently inside the lighthouse since the hurricane came through?
how many stair steps to top lookout...is it over 700 steps
What is the history behind the island?
See all questions


text: Venditore: poppyhoney (736) 100%, Luogo in cui si trova l'oggetto: Rayleigh, Spedizione verso: GB e molti altri paes, Numero oggetto: 201733906087 ITEM DESCRIPTION – Girls denim gilet. Age 10-11 Years. Studded collar. Never worn. Please check out my other auctions. I will combine and reduce postage costs for multiple purchases. PAYMENT - Payment by Paypal within 48 hours of the auction ending please. As a private seller unfortunately I can't offer refunds. Therefore, feel free to contact me if you need more information on the item before bidding. POSTAGE - Items will be sent by Hermes so if you live in a remote location there will be an additional postage cost. I will usually post all auctions on the next Tuesday or Friday after payment. I will post recorded signed for at an extra cost if required, please ask. Size: 10-11 Years, Brand: Denim & Co, Style: Gilets & Bodywarmers, Main Colour: Blue, Material: Denim


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Word Work Clip Art
This fun set of word work clip art features kids in various word work centers. This set contains 25 image files, which includes 13 color images and 12 black & white images in png. All images are 300dpi for better scaling and printing.
The Word Work clip art set includes:
- Boy using letter stamps
- Girl with magnetic letters on a cookie sheet
- Cookie sheet with magnetic letters
- Boy using dough
- Dough container
- Girl using letter tiles
- Letter tiles
- Girl using letter blocks
- Letter blocks
- Boy using a pocket chart
- Pocket chart
You will receive:
- 13 color png images
- 12 black & white png images
* All clip art sets are delivered via a zip file
This product is a digital download that is contained in a zip file and available to download immediately after purchase.
* Each clip art set come with a Standard License Agreement. A Standard License Agreement includes usage for personal, educational, and small business. Click here to learn more about the Standard License Agreement.


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I am an Independent Game Developer, and I am using Game Maker: Studio to develop games since 3-4 years. Since then I have developed many games, and I am still doing so. Currently I am making 3D games using the C# language in Unity Engine.
Through my course on Udemy, I share my knowledge of Game Maker: Studio and help people make games!
I have a beta project uploaded on Game Maker: Player called Nature Rescue, you can check it out.


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dork.major dork.
Reaching 1,243 in 2008 -- one day, one week, one mile at a time.
who knows...
Hi all,
I'm doing my first trail race on the 8th of September. The Montrail Run Like A Girl 8k in the Boston area. The event link is below. Most of my training has been on roads (some dirt country roads -- a lot in city streets and bikepaths) and so I'm wondering how to adjust my pace expectations. Any thoughts? I don't think this trail is going to be very technical (forest roads, fields, along beaver ponds) but since I've never done this before I'm not sure how to adjust my pace expectations. Any advice would be greatly appreciated, and if you're going to be there, let me know that too!!
Leslie Living and Running Behind the Redwood Curtain -------------
2017 Schedule:
Jan 8 - Clam Beach 8 3/4; Feb 18/19 - Hagg Lake 50k & 25k; May 7 - Ave of Giants HM; June 3 - Grasshopper Peak 30k (sweep duties); June 16/17 - Wild Rogue Relay; Jul 8 - Mt. Hood 50 (and maybe 50k next day - lottery); Sept 23 & 24 - Mt. Lakes 100 (first 100 - Yikes!!)
Trail Runner Nation


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American Army Seals off Target in Covert Operation
American Army seals effectively focused on a high esteem mission in Pakistan a couple of years prior. It was a mission led by Army commandos amidst night when the world was dozing all over. US Army seals had showed up before their objective and vanished inside no time. Target was accomplished and world come to know with stunning news in the media.
Commonly in past too, American Army has stunned the world with sudden results. Armed force Commandos in US Army as well as wherever on the planet are popular for such exercises. Such targets are just conceivable when officers are out there to make them have no trepidation of life.
Armed force commandos everywhere throughout the world are prepared after choice in intense standers. Now and then, it has seen that best Soldiers in US armed force are not ready to get chose in the Special gathering of commandos. Choice principles in American Army are dependably been extreme however with regards to exceptional units, a choice gets to be harder than normal.
The main Army Commandos unit on the planet that has given intense time to American Army was the Pakistani Commandos. Pakistan officers of unique units have vanquished US Army fighters in numerous war diversions held previously. As of late, a Pakistan warriors has won the honor of best expert sharpshooter shotter on the planet.
The Special units of Army commandos come to Pakistan to get the expert preparing. Pakistan Army and US Army have common instructional meetings and war diversions too. American Army and Pakistan Army remains at the highest point of world in practically every expert field.
Inside Story of Compound Raid in Pakistan from… by RealNewsTV


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Bristol-Myers Kidney Cancer Drug Boosts Survival in Big Studyby and
Patients on Opdivo lived months longer than those on Afinitor
Another trial finds Exelixis drug slows pace of same disease
A large study found that Bristol-Myers Squibb Co.'s drug Opdivo extends survival for people with advanced kidney cancer, a result that could provide new options for the hard-to-treat tumor.
The 821-patient study being presented Saturday at the European Cancer Congress in Vienna found that kidney cancer patients who had failed initial treatments lived more than five months longer if they took Opdivo, compared to taking a standard kidney cancer drug, Novartis AG's Afinitor. Patients on Opdivo also reported better quality of life, according to the results.
Bristol-Myers said it plans to apply for approval for the new use of Opdivo, which already is prescribed for lung cancer and melanoma and costs about $150,000 a year. Using the drug for kidney cancer patients could eventually add more than $1.3 billion of sales for New York-based Bristol-Myers, according to a Sept. 22 report from David Risinger, an analyst at Morgan Stanley.
"This is the first study that really shows that we're improving survival in patients" with kidney cancer, said Robert Motzer, an oncologist at Memorial Sloan Kettering Cancer Center in New York and the lead author of the study.
Of several newer drugs available for kidney cancer, almost all have been shown merely to slow tumor growth, Motzer said. Only one other drug has shown a survival benefit, and that drug was studied in a much narrower population.
New Class of Drugs
Opdivo is one of a new class of drugs that stimulate the immune system to attack cancers. It competes with Merck & Co.'s Keytruda, which is approved for advanced melanoma. The drugs are expected to be used in many different types of tumors and eventually become a mainstay of cancer care.
Patients who took Opdivo lived a median of 25 months, compared to 19.6 months for patients taking Afinitor, according to results that also are being published in the New England Journal of Medicine. In addition, 19 percent of Opdivo patients had serious side effects, compared to 37 percent who had bad side effects on Afinitor.
Common side effects with Opdivo in the study included fatigue, nausea and diarrhea. In previous trials, Opdivo has caused a variety of auto-immune effects, including severe lung inflammation in a number of cases.
Bristol-Myers announced on July 20 that it had stopped the trial early because the drug worked well. This is the first presentation of the full results from the company-sponsored study.
Thyroid Cancer Treatment
In a separate trial being presented at the meeting, a thyroid cancer drug from Exelixis Inc. also yielded positive results in kidney cancer patients who had failed other treatment. Interim results from the trial in 658 patients found that the drug, called Cometriq, slowed progression of the disease by 7.4 months, compared to 3.8 months for patients treated with Afinitor. Three out of five patients on Cometriq needed to reduce their doses due to side effects, compared to one in four people on Afinitor. Side effects of the Exelixis drug include hypertension, diarrhea and fatigue.
Novartis declined to comment directly on either trial. "We welcome all scientific insights that can improve outcomes for patients," the Swiss company said in an e-mail.
The Cometriq trial, sponsored by Exelixis, hasn't yet continued long enough to show definitely whether the medicine lengthens life, according to the results. Exelixis, a San Francisco-area biotech company, said it expects to complete its application for approval of the new use by the end of this year. The company also expects to file in Europe for the new use next year.
More than 60,000 Americans develop kidney cancer each year, and more than 14,000 die a year, according to estimates from the American Cancer Society. Risk factors include smoking and obesity.


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HUBBELL WIRING DEVICE-KELLEMS| Zoro #: G1057901| Mfr #: HBL3335C
Zoro #: G1057901Mfr. Part #: HBL3335C
$74.05 /ea
This item ships free. DetailsStandard ground shipping
Technical Specifications
|Zoro #: G1057901 Mfr #: HBL3335C
|Number of Poles: 3
|Amps: 30
|Material: Valox
|Standards: UL/CSA
|Item: Flanged Locking Receptacle
|Voltage: 125/250VAC
|Color: White
|Special Features: With Cover
|NEMA Rating: 4X, 12
|NEMA Configuration: Non-NEMA
|Mounting Type: Flush Mount
|Grade: Industrial
|Shrouded / Non-Shrouded: Non-Shrouded
|Phase: 3
|Wiring Style: Standard
|Number of Wires: 3
|IP Rating: 20
|Country of Origin (subject to change): United States
Product Description
Flanged Locking Receptacle, Shrouded / Non-Shrouded Non-Shrouded, Wiring Style Standard, Mounting Type Flush Mount, Grade Industrial, Material Valox, 30 Amps, Voltage 125/250VAC, Number of Poles 3, Number of Wires 3, Color White, Non-NEMA NEMA Configuration, NEMA Rating 4X, 12, IP Rating 20, Phase 3, Standards UL/CSA, Special Features With Cover


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60s throwback
Join date: Feb 201410 IQ
Our band is currently in negotiations to play poolside for 1000 fans or so at a hotel resort adjacent to the Desert Trip event at Coachella. It seems the band originally booked dropped out so we may be in. Being a warmup band for the Stones, McCartney, Roger Waters, Neil Young, and The Who sounds like it might be a kick. Will post pics if this thing really happens.
Sick! Keep that sounds like a crazy fun gig
Sick! Keep that sounds like a crazy fun gig


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Call (888) 651-9785
Let a private chauffeur transport you by luxury vehicle to Montserrat from Barcelona. You'll be accompanied by a personal guide for a 3-hour tour of the Santa María de Montserrat Abbey. Get to know the famed monastery and learn about the significance of this religious landmark, considered one of Spain's foremost pilgrimage sites. Upgrade your tour to a full-day itinerary with a winery or cava (Catalan sparkling wine) tour and tasting.
August 2016
The tour was outstanding and special. Angela was very knowledgeable.
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Panasonic KX-TG4312BX2 Cordless Landline Phone
Warranty Period:
1 Year Manufacturer's Warranty
Other Call Features:
Walkie-Talkie Mode, Paging, Call Share
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660 hrs
Other Menu Features:
Time Display and Menu Operation
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Panasonic KX-TG4312BX2 Cordless Landline Phone
PParul Aneja
The Panasonic Cordless model is a very portable, light in weight cordless phone. It has multiple features that include caller Id, alarm, date and time, etc It also includes hands free speaker. It has excellent voice clarity. Also, one can maintain a phone book which comprises of all the contacts. Moreover, there is a list of recently dialled numbers. So one can easily browse through the list to check the recently dialled calls. Speed dialling option is also available. There's also a provision for changing the ringtones and controlling the volume of the ringtone which is present in almost all the cordless phones . Though the designing is not that attractive, yet the phone serves the purpose.Read less Read more
RRitika Sarma
Starting from the design, the looks of this phone are obsolete and don't catch the eye. Secondly, the battery backup is not very good and can experience fast battery drainage at times. The price being charged for the features provided is too much, and the same money can buy something better. The range for this phone is good and can be used in a large area. However, the overall rating stays low.Read less Read more
this stylish cordless phoneis also used for paging it has a feature of conference calling also it has a good design and good sound quality it is a durable product with high ranges and comfortable in use it is east to setup with correct size and weight it is easy to manage it and its easy to clean it up it is a comfortable choice to go forRead less Read more
This Cordless is the best you can get but the only problem this has is the price. It's a lot more expensive than all the other cordless phones in this range. It has an alplanumerical keypad with Caller ID support and 5 hours talktime. It also has extra features like conference calls, etc. The best you can get!Read less Read more
Product Details
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1 Year Manufacturer's Warranty
In The Box
Sales Package
Handset and Base
Cordless Phone
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Call Features
Other Call Features
Walkie-Talkie Mode, Paging, Call Share
Power Features
Handset Battery
AAA Ni-MH Battery
Standby Time
5 hrs
660 hrs
Additional Features
Other Features
Any Key Answer, Call Share, 5.8 GHz Frequency
Memory Features
Phonebook Entries
50 Entries
Redial Memory
5 Redial
Convenience Features
Other Menu Features
Time Display and Menu Operation
Display Features
Display Type
1.4 inch Dot Matrix Display
Number of Display Lines
3 Display Lines
Other Display Features
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Aaya Nagar, Alaknanda, Alipur, Anand Lok, Anand Niketan, Anand Vihar, Ansari Nagar, Asaf Ali Road, Ashok Vihar Phase 1, Ashok Vihar Phase 2, Ashok Vihar Phase 3, Aurangzeb Road, Aurobindo Marg, Azad Market, Azadpur, Barakhamba Road, Bhikaji Cama Place, Chanakyapuri, Chander Nagar, Chandni Chowk, Chawri Bazar, Chhatarpur, Chittaranjan Park, Civil Lines, Connaught Place, Daryaganj, Defence Colony, Delhi Cantt., Delhi University-GTB Nagar, Dilshad Garden, Dr. Zakir Hussain Marg, Dwarka, East of Kailash, East Patel Nagar, Feroze Shah Road, Friends Colony, Geeta Colony, Geetanjali Enclave, Gole Market, Greater Kailash (GK) 1, Greater Kailash (GK) 2, Greater Kailash (GK) 3, Green Park, GTB Nagar, Gujranwala Town, Hauz Khas, Hauz Khas Village, IGI Airport, Inderlok, India Gate, IP Extension, ITO, Jail Road, Jama Masjid, Janakpuri, Jangpura, Janpath, Jasola, JNU, Jor Bagh, Kailash Colony, Kalkaji, Kamla Nagar, Kapashera, Karampura, Karkardooma, Karol Bagh, Kashmiri Gate, Khan Market, Khanpur, Khel Gaon Marg, Kirti Nagar, Krishna Nagar, Lajpat Nagar 1, Lajpat Nagar 2, Lajpat Nagar 3, Lajpat Nagar 4, Lawrence Road, Laxmi Nagar, Lodhi Colony, Lodhi Road, Mahipalpur, Majnu ka Tila, Malcha Marg, Malviya Nagar, Mandi House, Mansingh Road, Mathura Road, Mayapuri Phase 1, Mayapuri Phase 2, Mayur Vihar Phase 1, Mayur Vihar Phase 2, Mayur Vihar Phase 3, Mehrauli, MG Road, Delhi, Model Town 1, Model Town 2, Model Town 3, Moti Bagh, Moti Nagar, Mukherjee Nagar, Munirka, Najafgarh, Nangloi, Naraina, Narela, Nehru Place, Netaji Nagar, Netaji Subhash Place, New Friends Colony, Nizamuddin, Okhla Phase 1, Okhla Phase 2, Okhla Phase 3, Paharganj, Palam, Panchsheel Park, Pandara Road Market, Pandav Nagar, Parliament Street, Paschim Vihar, Patparganj, Pitampura, Pragati Maidan, Prashant Vihar, Preet Vihar, Pul Pehlad Pur, Punjabi Bagh, Qutab Institutional Area, Race Course, Raja Garden, Rajendra Place, Rajinder Nagar, Rajokri, Rajouri Garden, Rana Pratap Bagh, R K Puram, Rohini, Safdarjung, Sainik Farms, Saket, Sardar Patel Marg, Sarita Vihar, Sarojini Nagar, Satyaniketan, SDA, Shahdara, Shahpur Jat, Shakarpur, Shalimar Bagh, Shastri Park, Sheikh Sarai, Siri Fort Road, South Extension 1, South Extension 2, South Patel Nagar, Srinivas Puri, Subhash Nagar, Sundar Nagar, Tagore Garden, Tilak Marg, Tilak Nagar, Trilokpuri, Tughlakabad Institutional Area, Uday Park, Uttam Nagar, Vasant Kunj, Vasant Vihar, Vasundhara Enclave, Vijay Nagar, Vikas Marg, Vikaspuri, Vivek Vihar, Wazirabad, Wazirpur, West Patel Nagar, Yusuf Sarai, Zakir Nagar
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text:
Hello, we are Hawaii Dream/Jvid Productions and we are a team of professionals that specialize in filming, editing and preserving your most important occasions. We use state-of-the-art technology and equipment to capture and transform these occasions into a perfected video for you to share with your colleagues, friends and family.
We work with you and listen to your ideas and suggestions to make sure that you're supremely satisfied every step of the way.
Please view our projects and services and contact us so we can begin crafting your next video.
Did the videographer capture all important wedding moments? Candids? Interviews?
We are extremely disappointed with the lack of professionalism on Doug's part. Before the wedding and during the wedding, we had a great experience with Doug as a videographer. However, after the wedding, we were sent the wedding videos of some other couple's wedding instead of the videos for one of our events.
We contacted Doug about this matter, and he told us we would receive the correct videos. After a few months, he only sent us part of the video. When we contacted him again (multiple times), he assured us he would send us the rest. It has been several months since then, and we have not received the videos. He has stopped responding to all modes of communication (he will not respond to emails or pick up/return phone calls).
Favorite part of the wedding video? Anything beyond ceremony & reception?
"After searching online for local videographers, my fiancé (now husband :D) and I came across their website. What first caught my attention was the price. I had gotten quotes from other companies but because of a 2 hour travel time the quotes were not in our budget. We scheduled a meeting with Doug and got a great first impression of him and a reasonable quote. He is a really nice guy and a great people person. It was so easy to talk to him about the important memories we wanted him to film. On the day of the wedding, Doug was great. He was very professional and did a really good job of getting good footage without getting in the way. It was so great to look at our wedding video. Many times he recorded us just having a great time, and we had no idea he was even filming us! Jvid Productions did a fantastic job!!"
- Sandra Smith
Favorite part of the wedding video? Anything beyond ceremony & reception?
"There are so many amazing moments I either didn't know happened, or forgot about due to the commotion of the big day and I am so incredibly happy we had Doug to film all those special moments. They were incredibly professional, fun, and speedy. The edited video was perfect. You can really see the passion they have for their profession during the entire process and in the finished product. They were a joy to work with and I would recommend them to anyone!"
- Courtney Harper
Favorite part of the wedding video? Anything beyond ceremony & reception?
Hands down our experience in working with Doug was SUPERB. He won our hearts when we met him in person. He greeted us with a warm smile and a professional appearance. Doug was punctual, professional, friendly, and very personable. He was also very accommodating and flexible. We extensively researched various videographers and overall Dougs price was very reasonable. He truly went above and beyond. For example, he went on the rooftop of the hotel in order to capture the beautiful monuments of Washington, DC and there were a couple occasions when Doug came over to check on us during the reception. Moreover, Doug accompanied us to another location where we were having our pictures taken. During the drive, he was joking with us and sharing stories of his travels. Even though we had only known Doug for a brief period of time, we were comfortable exchanging stories with him and felt like we had know him for years. Our guests also complimented on how unintrusive Doug was during our wedding. We promise you that you wont regret choosing Doug as your videographer. We would highly recommend him to anyone looking for a videographer!
- Walena & Nabeel
Copyright © 2001-2017 OneWed Inc.
All Rights Reserved.
Copyright © 2001-2017 OneWed Inc. All Rights Reserved.


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Chicken Tortilla Soup
Chicken Tortilla Soup
with Guacamole Wontons
I love soup dumplings and have tried them everywhere from San Francisco to Singapore. I have learned that if you fill your dumplings with ingredients that are either frozen or chilled, you can fry them and have them remain cool despite a warm, crispy exterior. I would later see this theory proven time and time again by the brilliant chefs of California and Texas state fairs, where they fry everything from butter to soda. My guacamole dumplings are another such example and are a great addition to this soup.
- Est. Cooking Time: 60 min
- Allergens: Milk,Wheat
- Skill Level:
- Spice Level:
- Nutrition Facts
Suggested Pairings:
- Riesling
- Belgian Ale
- Garlic Cloves
- Wonton Wrappers
- Chili Powder
- Small Onion
- Crushed Tomatoes
- Jack and Cheddar Cheese
- Corn Tortillas
- Cilantro
- Jalapeño Pepper
- Lime
- Chicken Breast
- Chicken Stock Concentrate
- Oregano
- Avocado
- Fryer Thermometer
- Heavy Medium Saucepot
- Large Saucepot or Dutch oven
- Medium Bowl
- Pastry Brush (optional)
- Sheet Pan Lined with Foil
- Salt
- Black Pepper
- Olive Oil
- Vegetable Oil
- It is normal for chicken to have a slight odor due to the vacuum sealed packaging.
Tips & Techniques:
Exclusive video demonstrations of the skills you'll need to bring this meal to life.
You might also like...
Italian Chicken Cacciatore
Dinner for 2
Spicy Tomato-Jalapeño Chicken Breast
with Sautéed Green Beans
Dinner for 2
Indian Tikka Chicken
with Sautéed Broccoli
Dinner for 2


text:
Results 1 to 7 of 7
August 21st, 2002, 02:58 PM #1
I was just checking my webpages and noticed that 5 of the ones I checked were ranked between 5-7 PR. Then when I just went back they were back to 2-4. Is there a reason for this???
The OMG is for my post below!!!!
[ 08-21-2002: Message edited by: DaPackster ]
[ 08-21-2002: Message edited by: DaPackster ]
August 21st, 2002, 03:00 PM #2
- Join Date
- January 18th, 2005
- Posts
- 161
The update is happening as we speak. PR will fluctuate wildly untill the whole thing settles down in 4-5 days...
August 21st, 2002, 03:17 PM #3
Nick, thanks. I was wondering what was happening. I didn't see the other posts about the dance going on. Eeeks, it picked up a couple duplicate pages that I am switching over to a new host. Could be trouble for me.
August 21st, 2002, 03:24 PM #4
Ut OH!!!!   I just went through yesterday and completely overhauled one of my main pages. Actualy it is my main page. I just checked Google 2 and I have a number 2 spot for one of the hardest keywords to get. The cache is completely different. Should I copy the google cache and change it back. Like a fool I never saved an old version of my page before the update. :eek:
August 21st, 2002, 03:42 PM #5
- Join Date
- January 18th, 2005
- Location
- Palm Springs, CA
- Posts
- 866
I would copy the cache page. If Google likes a page then keep it.
Kip [img]/infopop/emoticons/icon_smile.gif[/img]
August 21st, 2002, 04:11 PM #6
Kip, thats what I was thinking. I wonder if it is ok to leave for a few days? I wonder if Google goes back before it does the main shuffle to check one more time for changes? Hopefully not. Who knows, Google usually hates me anyways so it probably won't stick anyways.
August 21st, 2002, 09:30 PM #7
- Join Date
- January 18th, 2005
- Location
- Palm Springs, CA
- Posts
- 866
Personally, I would change that thing back to what Google likes faster than you can say Google Dance. If Google likes a page I keep it there until it isn't feeling quite as friendly about it, then I make a few adjustments.
Good luck!
Kip [img]/infopop/emoticons/icon_smile.gif[/img]
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text:
Movie Trailers - August 1988
March 1988 April 1988 May 1988 June 1988 July 1988 August 1988 September 1988 October 1988 November 1988 December 1988 January 1989 February 1989
Popular trailers for movies released in August 1988
Total trailers found: 119
A Nightmare on Elm Street 4: The Dream Master Trailer (1988)
19 August 1988
Dream demon Freddy Krueger is resurrected from his apparent demise, and rapidly tracks down and kills the remainder of the Elm Street kids.
Police Story 2 Trailer (1988)
20 August 1988
The Hong Kong super-cop must stop a group of blackmailing bombers at the same time that the villains of the first Police Story are out for revenge.
Young Guns Trailer (1988)
12 August 1988
A group of young gunmen, led by Billy the Kid, become deputies to avenge the murder of the rancher who became their benefactor.
The Blob Trailer (1988)
05 August 1988
Remake of the 1958 horror sci-fi about a deadly blob which is the spawn of a secret government germ warfare project which consumes everyone in its path.
Married to the Mob Trailer (1988)
19 August 1988
Angela de Marco is fed up with her gangster husband's line of work and wants no part of the crime world.
The Last Temptation of Christ Trailer (1988)
12 August 1988
Jesus (Willem Dafoe), a humble Judean carpenter beginning to see that he is the son of God, is drawn into revolutionary action against the Roman occupiers by Judas (Harvey Keitel) -- despite his protestations that love, not violence, is the path to salvation.
Tucker: The Man and His Dream Trailer (1988)
12 August 1988
Based on a true story. Shortly after World War II, Preston Tucker is a dynamic engineer and an enthusiastic showman who envisions the car of the future.
Tin Toy Trailer (1988)
01 August 1988
Babies are hardly monster-like, unless you're a toy. After escaping a drooling baby, Tinny realizes that he wants to be played with after all.
Door on the Left as You Leave the Elevator Trailer (1988)
31 August 1988
Yan is a successful artist. One day he is waiting for his date Florence to turn up at his apartment. As the doorbell rings, he finds another young woman in underwear on his doorstep: his neighbor Eva! She tells him she locked herself out of her apartment and insists that he helps her.
Alice Trailer (1988)
03 August 1988
A memorably bizarre screen version of Lewis Carroll's novel 'Alice's Adventures in Wonderland'. The original story is followed reasonably faithfully, though those familiar with this director's other films won't be the least bit surprised by the numerous digressions into Svankmajer territory, living slabs of meat and all.
Mac and Me Trailer (1988)
12 August 1988
A Mysterious Alien Creature (MAC) escaping from nefarious NASA agents, is befriended by a young boy in a wheelchair.
Summertime Blues Trailer (1988)
18 August 1988
Those Lemon Popsicle boys are at it again!..Naughtier than ever in...
1969 Trailer (1988)
18 August 1988
Two rebellious youths, Ralph and Scott, find themselves struggling with adulthood as the Vietnam War rages.
A Few Days with Me Trailer (1988)
14 August 1988
Martial (Daniel Auteuil) is discharged from a mental insitution where he spent a few years due to a serious nervous breakdown.
Vibes Trailer (1988)
05 August 1988
Medium Sylvia Pickel and psychometrist Nick Deezy meet at a psychic research facility in New York. Not long after, they're contacted by Harry Buscafusco, who offers them $50,000 to find his lost son in South America, in the heart of Incan territory where they discover an ancient mystical secret, and each other.
Clean and Sober Trailer (1988)
10 August 1988
Hotshot real estate salesman Daryl has a bad cocaine habit. After embezzling his company's money, he wakes up next to a girl who overdosed.
The Thin Blue Line Trailer (1988)
28 August 1988
Errol Morris's unique documentary dramatically re-enacts the crime scene and investigation of a police officer's murder in Dallas.
Betrayed Trailer (1988)
26 August 1988
An FBI agent (Debra Winger) falls in love with a white supremacist (Tom Berenger) whose group she infiltrates.
Hero and the Terror Trailer (1988)
26 August 1988
Danny O'Brien is back in action fighting the notorious Simon Moon, also known as The Terror. Three years earlier O'Brien had single-handedly captured The Terror and was called Hero by the people of L.
Gotham Trailer (1988)
21 August 1988
A New York private eye searches for a socialite who supposedly drowned 10 years before.
Quiet Thunder Trailer (1988)
11 August 1988
A bush pilot and a senator's wife try to survive in the jungles of Africa after witnessing an assassination.
Homeboy Trailer (1988)
24 August 1988
Johnny Walker is a cowboy and a boxer. He is very shy and a bit of a fool. He is in love with Ruby, but he cannot tell her.
Stealing Home Trailer (1988)
26 August 1988
Billy Wyatt (Harmon), a former high school and minor-league baseball baseball player receives a telephone call from his mother revealing that his former child-sitter, and later in his teens, his first love, Katie Chandler (Foster), has died.
Search and Destroy Trailer (1988)
31 August 1988
SEARCH AND DESTROY (1988) - Terrorists destroy a chemical factory. A virus is released that infects the people of the town with a strange psychosis.
Only We Can Love Like This Trailer (1988)
12 August 1988
Anneli is soon to be 17 years old. She lives alone with her mother and can barely remember her father who left the family 10 years ago.
Put More Blood Into the Music Trailer (1988)
06 August 1988
PBS produced documentary about Sonic Youth at the height of their powers in 1988
The Mayflower Voyagers Trailer (1988)
21 August 1988
The Peanuts characters take the voyage to the new world. When they get there, they learn about disease, planting, building, and Squanto shows them how to get corn (or maize, as he calls it) to grow.
Strike Commando 2 Trailer (1988)
17 August 1988
Michael Ransom's Vietnam squad leader, Vic Jenkins, is captured by terrorists demanding ten million dollars worth of diamonds in return for his release.
Sherlock Holmes: The Hound of the Baskervilles Trailer (1988)
31 August 1988
Sir Charles Baskerville dies on the moor under mysterious circumstances and rumors abound about a demonic hound.
Cycling the Frame Trailer (1988)
30 August 1988
This is a short film (30min), part documentary part art-film, about the Berlin Wall back in 1988 when West Berlin was still an isolated fiefdom of the capitalist west, located 125 miles behind the Iron Curtain in communist eastern Europe.
Top Line Trailer (1988)
16 August 1988
A down-on-his-luck author makes the find of a lifetime in the remote Columbian jungle, in the form of an alien spacecraft .
Last Platoon Trailer (1988)
21 August 1988
After ex-cop and highly decorated war hero Chet Costa's Vietnamese girlfriend disappears, he accepts a risky mission: he shall blow up a bridge at the Vietnamese border to close the Vietcong's line of communication.
The Stick Trailer (1988)
23 August 1988
A squad (stick) of South African soldiers is sent into the bush to track down some rebel fighters when things take a supernatural turn.
Moonnam Pakkam Trailer (1988)
28 August 1988
Thampi retired as an accountant from Indian Railway in mid-1980s following his son's death leads a solitary life in his ancestral house.
Ollie Hopnoodle's Haven of Bliss Trailer (1988)
05 August 1988
From Jean Shepherd (A Christmas Story) comes the story of 14 year old Ralphie's first job, and the family vacation the family is planning to a rustic fishing cabin on Lake Michigan.
Katinka Trailer (1988)
24 August 1988
A new foreman, Huus, arrives in a sleepy Danish village, much to the delight of the unmarried women there.
La travestie Trailer (1988)
03 August 1988
Lone Wolf Trailer (1988)
16 August 1988
A struggling rock band and other college students cope with the trials and tribulations of their young lives and find themselves in the role of determining what exactly is behind the recent full-moon killings which have been attributed to wild dogs.
The Perfect Murder Trailer (1988)
03 August 1988
Police Inspector Ghote lives a middle-class life in Bombay along with his wife, Pratima. He has been employed with the Bombay Police for many years.
The Rescue Trailer (1988)
05 August 1988
A team of Navy Seals are sent to destroy a disabled submarine so it will not fall into the "wrong" hands.
Desert Rats Trailer (1988)
07 August 1988
Series pilot about a local rebel who is appointed sheriff of a small Arizona town.
Rodney Dangerfield: Nothin' Goes Right Trailer (1988)
10 August 1988
Rodney Dangerfield hosts a showcase of comics including himself, Robert Schimmel, Bill Hicks, and Andrew Dice Clay, among others.
Seven Days' War Trailer (1988)
13 August 1988
Junior-high students are restrained by their teachers and parents... One day, they plan a rebellion..
Aparan Trailer (1988)
08 August 1988
Aparan (The Impostor) is a 1988 Malayalam mystery film written and directed by P. Padmarajan, starring Jayaram, Mukesh, Shobana, Parvathy Jayaram and Madhu.
Hot to Trot Trailer (1988)
26 August 1988
Fred P. Chaney receives as inheritance after the death of his mother a speaking horse that also has good knowledge about the stock-market.
Marana Mrudangam Trailer (1988)
04 August 1988
Marana Mrudangam is action drama based movie in which, Johny (Chiranjeevi) runs a small time restaurant with his partner (Nagendra Babu).
Janaki Ramudu Trailer (1988)
19 August 1988
Janaki Ramudu
FleshEater Trailer (1988)
17 August 1988
A group of teenagers, taking a nocturnal hayride come across the grave of a man. Little did they know that this deceased man is a zombie.
The Reader Trailer (1988)
17 August 1988
A multi-faceted film based on Raymond Jean's novel "La Lectrice". Constance (Miou-Miou) reads the novel aloud in bed to her lover.
Invasion Earth: The Aliens Are Here Trailer (1988)
22 August 1988
Insect-like aliens invade a small town. The local teenagers have been watching a sci-fi marathon in the local theater, and from those films they get ideas on how to fight the creatures.
The Black Arrow Trailer (1988)
16 August 1988
Hope and Pain Trailer (1988)
06 August 1988
Directed by Yôji Yamada
WWE SummerSlam 1988 Trailer (1988)
29 August 1988
SummerSlam (1988) was the first annual SummerSlam professional wrestling pay-per-view event. It was produced by the World Wrestling Federation and took place on August 29, 1988 in Madison Square Garden, located in New York, New York.
Tupla-Uuno Trailer (1988)
26 August 1988
Fever Trailer (1988)
26 August 1988
A policeman discovers a suitcase full of drug money while on duty, meanwhile his wife is having an affair with another man.
Heart to Hearts Trailer (1988)
25 August 1988
The first of the three titles in the screwball Hong Kong "Hearts" series. Alex, a creative director, temporarily fills in as conductor for a choir.
El virus del poder Trailer (1988)
30 August 1988
Tomorrow Trailer (1988)
13 August 1988
On August 9, 1945, the Americans dropped an atomic bomb on Nagasaki. This film, based on a story by Mitsukaru Inoue, describes the daily life of people in Nagasaki the day before that fateful event.
Envoyez les violons Trailer (1988)
03 August 1988
Aryan Trailer (1988)
01 August 1988
Aryan movie is all about Devanarayan (Mohanlal) who is a Brahmin priest who is falsely accused of theft and forced to leave his village in Kerala.


text:
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APs are rebooting regularly. Cisco switch is providing POE to these APs. Is there any known issue with this setup?
by Shyam_Moolayilk
Question: APs are rebooting regularly. Cisco switch is providing POE to these APs. Is there any known issue with this setup?
Product and Code:
All controller and All Code. Cisco 3550PWR switches and 12.1.(19) code
APs connected to Cisco POE switches reboots regularly, the reason is that when the AP comes up for the first time after it
downloads the code from the controller it reboots. Now the Cisco switch thinks that the AP is not there anymore on the port and shuts the power to that port.
The workaround for this issue is to increase the time the Cisco waits before shutting down a port. The following command can be used to do the same.
power inline delay shutdown 15 initial 25
This only applies to the Cisco3550PWR. All cisco switches released after this model do not require any special config. The Cisco IOS on the 3550 has to be at least 12.1.(19) or higher for that command to work.


text:
Private In-Home and Online Chemistry Tutoring in Nashua, NH
Receive personally tailored Chemistry lessons from exceptional tutors in a one-on-one setting. We help you connect with in-home and online tutoring that offers flexible scheduling and your choice of locations.
Recent Tutoring Session Reviews
"The student and I had a ton to cover today. We focused on stoichiometry and reaction problems, provided by his teacher. We are working on remembering his stoichiometric relationships, but after a few problems, he got a better handle. I think he definitely needs to keep practicing before tomorrow, but he has a solid base."
"Today we finished chapter 4 on reactions in aqueous solutions. We reviewed definitions from earlier in the chapter (strong/weak acids/bases, redox) and did practice problems involving molarity (concentration), gravimetric analysis, and titration. The student completed the homework I assigned last time, which we reviewed together, and he said he thought it was the right amount and the practice helped. He has no problems understanding mass/moles/concentration in acid/base neutralization reactions. I am happy with the progress we are making and on Friday we plan to review ideal gas behavior and focus on deviations from ideal gas behavior."
"Today the student completed a homework assignment on electron orbital configurations and another homework assignment on atomic structure (atomic number, mass number, charge and what that means with regard to the number of protons, neutrons, and electrons), isotopes and computing mean atomic masses. Then the student completed a lab activity on precipitating out compounds in sequence given the solubility rules."
"We went through her worksheet for gas laws. I explained them to her and asked her to solve different questions to make sure she understood them. She understood them and she was able to do most of the questions by herself. I also summarized the important parts for her so she can have them handy. She is doing well."
"We covered acids and bases, balancing oxidation-reduction reactions, and identifying reaction types. The student had a clear idea of what she wanted to cover in the session and seemed very engaged with the material. More practice identifying reaction types might be useful."
"The student and I worked on test prep; going over all the subjects and identifying gram->gram conversions and limiting reactants / percent yield problems as the most pressing. We then worked through some problems in the book, and I identified 12 or so practice problems for her to work on before my visit tomorrow. She will also try to get an example exam, so we can do the final test prep tomorrow, before her Thursday exam"
Nearby Cities:Framingham Chemistry Tutoring, Westminster Chemistry Tutoring, Providence Chemistry Tutoring, Worcester Chemistry Tutoring, Hartford Chemistry Tutoring, Warwick Chemistry Tutoring, Cranston Chemistry Tutoring, Woonsocket Chemistry Tutoring, Leominster Chemistry Tutoring, Fitchburg Chemistry Tutoring, Marlborough Chemistry Tutoring, Norwich Chemistry Tutoring, Franklin Chemistry Tutoring, New Hampshire Chemistry Tutoring, Rhode Island Chemistry Tutoring
Nearby Tutors:Framingham Chemistry Tutors, Westminster Chemistry Tutors, Providence Chemistry Tutors, Worcester Chemistry Tutors, Hartford Chemistry Tutors, Warwick Chemistry Tutors, Cranston Chemistry Tutors, Woonsocket Chemistry Tutors, Leominster Chemistry Tutors, Fitchburg Chemistry Tutors, Marlborough Chemistry Tutors, Norwich Chemistry Tutors, Franklin Chemistry Tutors, New Hampshire Chemistry Tutors, Rhode Island Chemistry Tutors


text:
Nice to meet you on Audiko.net! A perfect occasion to download a free AEROC ringtones for your iPhone or mobile. Take your favorite slice from your favorite piece of art - Aerocity - Love Lost, Passages, Stranger, or any other track. You are free to obtain your tone both in the MP3 and m4a formats, played by both mobile phones and iPhones. Just select a track, and choose free AEROC ringtones download option.
Download AEROC Ringtones
Top free ringtones from remixes
|Love Lost
|Aerocity - Love Lost


text:
- Location:
- Elgin, IL, USA
- Price:
- Contact Seller
- Manufacturer:
- Associated Research
- Model:
- 7710
- Condition:
- used
- Condition info:
- Refurbished
- Subcategory:
- Insulation testers
- Subcategory 2:
- Electronic test equipment
Associated Research 7710 Features:
Patented SmartGFI® safety circuit protects the operator from shock hazards
50 memories that can be stored and recalled. Multi-function testers include 8 steps per memory
RAMP HI and CHARGE LO systems for more effective DC Hipot testing
500VA testers available for Higher Current Hipot test applications
Meets 200 mA short circuit requirements
Up to 12kV AC or DC Hipot testing for manufacturers with higher voltage testing requirements
RS-232/USB or GPIB automation interfaces available
Autoware Testing Software available for complete Automation Control
The Associated Research 7710 is a 12 KV AC Hipot tester that is designed to provide high DC output voltage for cable, wire, harness and electrical component testing. This easy-to-use bench top tester offers numerous features for safe and efficient testing. These features include our patented SmartGFI safety circuit and our patented CHARGE-LO and RAMP-HI systems. In addition , the 7710 can also be used in tandem with our Autoware control software.
Get email updates for Associated Research 7710


text:
I will hand Draw Zen tangle Patterns letters or names
About This Gig
Order Details
1 day delivery
Draw black and white zentangles
I will draw zentangles such as mandalas, doodled names, letters or anything. I will charge $10each
- High Resolution
Frequently Asked Questions
Will you be sending a black and white drawing or colored?Black and white. I can add a solid colored background or send you a vectorized drawing instead of a jpeg if you want.


text:
Sail the Schooner Heron for a fun and informative look at the history of Rockport Harbor. Sail across Penobscot Bay to see the seals and seabirds on Robinsons Rock, birthplace of Andre the Seal. Enjoy the thrill of sailing a classic wooden schooner.
Elegant, yet comfortable, 14 room bed and breakfast located in the heart of MidCoast Maine.
Come home to Freeport! Centrally located: walk to LL Bean, outlet shopping & restaurants. Homemade breakfast. 24 hour coffee & tea. A/C, WiFi. Non-smoking.
Maine Outdoors is here to help you create great outdoor memories! Everything that you need is provided on every trip and we specialize in families or beginners.
LedgeLawn,located on an Atlantic estuary is the perfect spot for canoeing and sea kayaking. Nestled between an Audubon bird sanctuary, and this Atlantic inlet, LedgeLawn is a wonderful place for kids or just to relax and live life at a slower pace.
A hundred year old inn on a hill overlooking Linekin Bay on the coast of Maine. 16 rooms. All but one overlook the water, big wrap around porch, Private baths, a full breakfast by a Culinary Institute trained chef.
All suites inn, located on the harbor in the quiet village of Southwest Harbor , Maine.
The Maine Windjammer Association's nine historic ships offer exciting 3- to 6-day sailing vacations along the Maine coast from May to October. Includes great food, spectacular scenery, abundant wildlife and time to explore islands and villages.
Rockport, Maine visitors guide to accommodations, adventure travel, unique tours and rentals. Check out top Rockport, Maine getaways & destinations. Rockport, Maine vacations, packages & travel spots. Visit Rockport, Maine !.