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Quinolinic acid toxicity on oligodendroglial cells: relevance for multiple sclerosis and therapeutic strategies
Gayathri Sundaram1, 2,
Bruce J Brew1, 3,
Simon P Jones1,
Seray Adams2, 4,
Chai K Lim1, 2, 4 and
Gilles J Guillemin1, 2, 4Email author
Journal of Neuroinflammation201411:204
© Sundaram et al.; licensee BioMed Central. 2014
Received: 1 September 2014
The excitotoxin quinolinic acid, a by-product of the kynurenine pathway, is known to be involved in several neurological diseases including multiple sclerosis (MS). Quinolinic acid levels are elevated in experimental autoimmune encephalomyelitis rodents, the widely used animal model of MS. Our group has also found pathophysiological concentrations of quinolinic acid in MS patients. This led us to investigate the effect of quinolinic acid on oligodendrocytes; the main cell type targeted by the autoimmune response in MS. We have examined the kynurenine pathway (KP) profile of two oligodendrocyte cell lines and show that these cells have a limited threshold to catabolize exogenous quinolinic acid. We further propose and demonstrate two strategies to limit quinolinic acid gliotoxicity: 1) by neutralizing quinolinic acid's effects with anti-quinolinic acid monoclonal antibodies and 2) directly inhibiting quinolinic acid production from activated monocytic cells using specific KP enzyme inhibitors. The outcome of this study provides a new insight into therapeutic strategies for limiting quinolinic acid-induced neurodegeneration, especially in neurological disorders that target oligodendrocytes, such as MS.
Oligodendrocyte
Excitotoxicity
Quinolinic acid (QUIN) is a downstream metabolite produced through the kynurenine pathway (KP) of tryptophan metabolism [1],[2]. In physiological conditions, QUIN is present in nanomolar concentrations and used as substrate by cells to synthesize the essential co-factor nicotinamide adenine dinucleotide (NAD+). However, during neuroinflammation, the KP can be chronically or acutely activated through the induction of one of its initial enzymes, indoleamine 2,3-dioxygenase (IDO-1). QUIN is then produced in excess and can kill brain cells including neurons, astrocytes and oligodendrocytes by at least six different mechanisms [2],[3]. QUIN has been implicated in several neurological diseases, including multiple sclerosis (MS) [4],[5]. The involvement of the KP in neurological diseases is complex, as it revolves around the metabolic balance between excessive production of neurotoxic metabolites, such as QUIN, and neuroprotective compounds, such as kynurenic acid (KYNA) [6],[7].
At the cellular level, QUIN is cytotoxic for most of the brain cell types in vitro at micromolar concentrations. Cammer [8],[9] showed that exposure to 1 mM of QUIN induces cell death in rat oligodendrocytes [8],[9]. Similar toxic effects are also observed in primary human astrocytes and neurons at pathophysiological concentrations of 150 nM [10], and more recently in motor neurons at concentrations of 100 nM [11]. Furthermore, this effect can be abolished by using antagonists of the N-methyl-D-aspartate (NMDA) receptor - such as memantine, MK801 and AP-V - implying excitotoxicity as the main mechanism inducing cell death [10],[11]. Current evidence suggests only monocytic lineage cells have the ability to produce QUIN [12],[13]. Brain cell types, including neurons, astrocytes, pericytes and endothelial cells are likely to uptake QUIN and catabolize it [14]-[17].
The function of the KP in oligodendrocytes remains to be investigated, although an earlier study demonstrated that IDO-1 and tryptophan 2,3-dioxygenase (TDO-2) are not expressed in human primary oligodendrocytes [5]. This potentially has strong implications for MS pathology. The lack of these two KP regulatory enzymes in oligodendrocytes is associated with a higher cell susceptibility to allogenic T-cell challenge, since IDO-1 plays a crucial role in immune regulation - particularly in suppressing T cell proliferation [18]. The KP profile has been shown to be altered in both MS patients and in experimental autoimmune encephalitis (EAE) mouse models [19]-[21]. Rejdak et al. [19] reported that the concentration of the neuroprotective KYNA in the cerebrospinal fluid (CSF) of MS patients is decreased during the remission stage. This indicates that the KP stays inclined towards neurodegeneration long after inflammation. We hypothesise that in MS the sustained activation of the KP in the CNS leads to an unbalanced production favoring neurotoxic over neuroprotective intermediates.
This study investigates the profile of the KP in oligodendrocytes and characterizes to what extent KP metabolism changes during immune challenge. We have also identified novel approaches to protect oligodendrocytes against pathophysiological concentrations of QUIN. The outcomes of this study could have significant implications for potential therapeutic strategies in MS and other neurodegenerative diseases that involve KP dysregulation with overproduction of QUIN.
Cell cultures
Mouse oligodendroglial cell lines (N19 and N20.1) were kindly provided by Dr. A T Campagnoni (University of California, Los Angeles, CA, USA). These cells were conditionally immortalized oligodendrocytes isolated from normal and shiverer primary mouse brain cultures through the use of the retroviral vector pZIPSVtsA58 encoding immortalizing thermolabile simian virus 40 large T antigen that allows for clonal selection by conferring neomycin (G418) resistance [22]. N19 and N20.1 were chosen for their differences in developmental maturity along the oligodendrocyte lineage. N19 represent oligodendroglial cells with a relatively immature or precursor phenotype, whereas N20.1 represent a more mature phenotype. Oligodendroglial cells were cultured in accordance with the previously published method [22],[23]. Briefly, cells were seeded and grown in flask coated with poly-L-lysine to confluence at 34°C, the permissive temperature, in DMEM/F12; Invitrogen, Melbourne, Australia) supplemented with 10% FBS, 100 μg/ml of gentamicin, 100 μg/ml of G418 (Invitrogen, Melbourne, Australia) and then shifted to 39°C, the non-permissive temperature that leads to 'differentiated' state for 72 hours. In all the experiments, during incubation at 39°C, the medium was replaced by fresh DMEM/F12 with 1% FBS and maintained for 7 days before treatments.
The BV2 cell line was kindly donated by Dr. T Kuffner (St Vincent's Center for Applied Medical Research, Sydney, Australia). The culture method for the BV2 murine microglial cells was adapted from Laurenzi and Arcuri et al. [24]. Briefly, BV2 cells were maintained in DMEM supplemented with 10% FBS, Glutamax and antibiotic-anti-mycotic solution. The mouse macrophage cell line RAW264.7 was kindly donated by Prof. Nicholas Hunt (University of Sydney). The RAW264.7 cells were cultured based on the method adapted from Watts and Hunt et al. [25].
The study conditions were: (1) unstimulated; (2) IFN-γ stimulated oligodendroglial cell lines (N19 and N20.1); and (3) IFN-γ-treated macrophage cell line, RAW 264.7 as a positive control, since macrophages are known to express all the KP enzymes [14]. The study design included 3 time points: 24, 48 and 72 hours based on previous similar studies showing peak mRNA expression of KP enzymes at 24 hours, whilst production of KP metabolites peaks at 72 hours post-treatment (data not shown) [15],[16].
Characterization of KP enzymes by quantitative RT-PCR
The qRT-PCR protocol used has been previously described [26]. The primer sequences used to detect mouse KP enzymes are summarized in Table 1. Primers were obtained from several sources: designed by Dr. Fabrice Magnino (PCR/qPCR specialist Integrated Sciences Pty Limited), a gift from collaborating researchers, Primer Bank (http://pga.mgh.harvard.edu/primerbank/) or designed by using Primer BLAST (web-based NCBI primer designing tool: http://www.ncbi.nlm.nih.gov/tools/primer-blast/) or Primer3 designing program (http://primer3.sourceforge.net/). RPL13 was used as the endogenous reference gene. Data are presented as fold differences in gene expression normalized to RPL13 and relative to the untreated control. Differences in the relative expression of each gene were analyzed using Student's t-test. Statistical significance was accepted at P < 0.05. Reported values are mean ± SE of triplicate samples.
Primer sequences for qRT-PCR
Primer sequence
Indoleamine 2,3-dioxygenase
IDO1
TGT GAA TGG TCT GGT CTC
CTG TGC CCT GAT AGA AGT
Tryptophan 2,3-dioxygenase
TDO2
BC018390.1
TGC TCA AGG TGA TAG CTC GGA
AGG AGC TTG AAG ATG ACC ACC A
Kynurenine aminotransferase 1
GCT TTT CAG CAG GCT ACC AC
CCA CTG TCA CCA GCA CAT TC
AF072376
GAA CTT CTG TCC TGG CTA A
CTT GAT TGG GTG GGT AGT
Kynureninase
KYNU
GAG CAG AGG AGC GTG GCT GC
GAA CAG GGG CCA CGC GGA TG
3-hydroxyanthranilate 3,4-dioxygenase
3HAO
TTC AGC CTC ATT GCA TCT
GAC AGT GTA GGG CTA TGG
Kynurenine 3-monoxygenase
GGT CGC CTT CAC CAG AAT AA
ATC CAG GCA GGT CTT CTC AA
Aminocarboxymuconate semialdehyde decarboxylase
ACMSD
GAA TAA ATG CTG ACC CAA CA
TTC ATC CAT CCT TCC AGA C
Quinolinate phosphoribosyl transferase
QPRT
CTG CTC CAA GTC ACC ATG
CAG AAC CCC AGG AGA TTT
Ribosomal protein L13
RPL13
GAG GTC GGG TGG AAG TAC CA
TGC ATC TTG GCC TTT TCC TT
Immunocytochemistry
To characterize the KP in both N19 and N20.1 cell lines, cells were cultured onto slide flasks (Nunc, Rochester, NY, USA) at 34°C and when 70% confluence was reached they were incubated at 39°C for 72 hours. After 3 days, the cells were either left untreated or treated with 100 IU/ml of IFN-γ for 24 hours. The cells were then stained for indoleamine 2,3-dioxygenase 1 (IDO-1) (Biolegend, San Diego, CA, USA), tryptophan 2,3-dioxygenase (TDO-2; generous gift from Dr. CL Miller John Hopkins University, Baltimore, MD, USA), kynurenine 3-monooxygenase (Abcam, Cambridge, UK), QUIN (Millipore, Bedford, MA, USA), picolinic acid (PIC; Abcam, Cambridge, UK), 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP; Millipore, Bedford, MA, USA) and myelin basic protein (MBP; Abcam, Cambridge, UK) according to previously described protocol [15],[16]. The antibodies are summarized in Tables 2 and 3. Untreated cells were stained and used as experimental control for comparison. Experiments were performed in triplicate and independently validated with three biological replicates.
Summary of primary antibodies
Myelin basic protein (MBP)
Monoclonal - IgG
2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP)
Vimentin (VIM)
Glial fibrillary acidic protein (GFAP)
Dako
Indoleamine 2,3-dioxygenase 1 (IDO1)
Tryptophan 2,3-dioxygenase (TDO)
Kynurenine 3-monooxygenase (KMO)
Quinolinic acid (QUIN)
Picolinic acid (PIC)
Summary of secondary antibodies
Alexa Fluor 488 (green)
Goat anti-mouse IgG
Goat anti-rabbit IgG
Alexa Fluor 594 (red)
Quantification of KP metabolites
Tryptophan, kynurenine, and kynurenic acid were measured using high performance liquid chromatography (HPLC) while picolinic acid and QUIN were concurrently measured using gas chromatography/mass spectrometry (GC/MS) as previously described [11]. Results of tryptophan and kynurenine were presented as kynurenine/tryptophan (K/T) ratio, a reliable measure of KP activation. Production or degradation of the KP metabolites was calculated by subtracting the values of post- and pre-treatment concentrations of the KP metabolites present in the culture medium.
QUIN toxicity and cell protection assays
QUIN toxicity and cell protection assays against QUIN were determined by calculating the percentage of cell survival rate in comparison to untreated controls using CytoTox 96® Non-Radioactive cytotoxicity assay kit (Promega, Madison, WI, USA). Experiments were divided into four parts:
QUIN toxicity on oligodendrocytes: to assess the QUIN toxicity, oligodendroglial cells were challenged with varying concentrations of exogenous QUIN up to 4 μM over 3 time points (24, 48 and 72 hours) and QUIN toxicity assessed. These data were used to determine the LD50 of QUIN toxicity and select the appropriate treatment period in subsequent QUIN antagonism experiments (Parts C and D).
QUIN uptake assays: to further demonstrate the uptake of QUIN in oligodendrocytes, cells were grown in slide flasks (Nunc, Rochester, NY, USA) and treated with LD50 dose described in Part A for 0, 30, 60 and 90 minutes using protocol adapted from [28]. QUIN uptake was then visualized using immunocytochemistry as described previously [11],[28].
Neutralization of QUIN with an anti-QUIN monoclonal antibody (mAb): to fully assess the potential of neutralizing QUIN toxicity with an anti-QUIN mAb, we subjected the oligodendroglial cells to 2 different conditions: 1. treated directly on oligodendroglial cell lines with exogenous QUIN followed by varying concentrations of QUIN-mAb with the following three conditions: (a) pre-treatment with QUIN (QUIN-PRE) for 72 hours at LD50 concentration followed by the QUIN-mAb for 30 minutes; (b) pre-treatment with anti-QUIN mAb for 30 minutes followed by QUIN (QUIN-POST) at LD50 concentration for 72 hours and; (c) concomitant treatment with QUIN and the anti-QUIN mAb (QUIN + QUIN mAb) together for 72 hours. 2. treated with IFN-γ-treated BV2 cells supernatant (endogenous QUIN) on oligodendroglial cell lines followed by varying concentrations of QUIN mAb. Cell death was then determined by measuring lactate dehydrogenase (LDH) in the culture supernatant.
Inhibition of QUIN production with IDO-1 inhibitors: to imitate QUIN production during inflammation and immune activation, BV2 cells were stimulated with IFN-γ for 24 hours to induce pathophysiological concentrations of QUIN production. Oligodendrocyte cell line cultures were then exposed to this QUIN-containing BV2 culture supernatant for 72 hours and assessed for QUIN toxicity. Further, the QUIN-producing BV2 cells were challenged with 4 specific IDO-1 inhibitors namely, 1-methyl-D-tryptophan (D-1MT), 1-methyl-L-tryptophan (L-1MT), 1-methyl-D-tryptophan (DL-1MT) and berberine (5,6-dihydro-9,10-dimethoxybenzo[g]-1,3-benzodioxolo[5,6-a]quinolizinium) for 30 minutes to block QUIN production as a potential therapeutic strategy to alleviate QUIN toxicity during neuroinflammation.
Results are expressed as mean ± SE. Differences between treatment groups for RT-PCR, GC/MS and HPLC data were analyzed using Student's t-test. QUIN toxicity study results were compared using linear and multiple regression analysis. QUIN mAb treatment results were analyzed using the Mann-Whitney test. In all experiments a P-value < 0.05 was considered significant.
KP profiling in oligodendroglial cell lines N19 and N20.1
We found that both mice oligodendrocytic cell lines N19 and N20.1 have a functional KP. This is evident at gene, protein and metabolite level in N19 (Figure 1a-d) and N20.1 (Figure 1e-h). However, the profile of KP expression in N19 was significantly different from N20.1 (see Additional files 1 and 2). By comparing the levels of mRNA expression for the KP enzymes, after normalization with the endogenous reference gene RPL13, we showed that N20.1 had a much higher level of expression for TDO-2 (P < 0.001) than N19 (Figure 1a and e). This result was validated at protein level using immunocytochemistry (Figure 1b and f). Both cell lines showed increased mRNA expression for TDO2 when treated with pro-inflammatory cytokine IFN-γ. In N20.1 cells most KP enzymes were up-regulated by IFN-γ, except ACMSD (see Additional file 1). As expected, treatment with IFN-γ led to an increased KP activation shown by the increased K/T ratio in both cell lines (Figure 1c and g). Interestingly this increase was due to an up-regulation of TDO-2 rather than IDO-1 gene expression in N19.
Characterization of the kynurenine pathway (KP) in N19 (a-e) and N20.1 (e-h) oligodendroglial cell lines. Levels of TDO-2 transcripts in (a) N19 and (e) N20.1 cells in differentiated state, with or without IFN-γ treatment. N19 cells (b) and N20.1 cells (f) showed positive TDO-2 cytoplasmic staining after 24 hours IFN-γ treatment. KP metabolic profile of oligodendroglial cell lines K/T ratio in N19 (c) and N20.1 (g) and kynurenic acid (KYNA), picolinic acid (PIC) and quinolinic acid (QUIN) profiles of N19 (d) and N20.1 (h) treated with or without IFN-γ. TDO-2, tryptophan 2,3-dioxygenase; K/T ratio, kynurenine and tryptophan ratio as a measure of IDO-1/TDO-2 activity; KYNA, kynurenic acid; PIC, picolinic acid; QUIN, quinolinic acid. Values are expressed as mean ± SE. ***P < 0.001, **P < 0.01 and *P < 0.05. Student's t-test.
KAT-1 and KAT-2 expression in both N19 and N20.1 suggests that oligodendrocytes are capable of producing the neuroprotective metabolite KYNA. Indeed, KYNA was detected in the culture media of both oligodendrocytic cell lines following background subtraction (serum supplemented media). Although there was a significant increase in KAT-1 and KAT-2 gene expression in N20.1 cells (see Additional file 1) following IFN-γ treatment, there was no significant difference in the levels of KYNA produced (Figure 1d and h). ACMSD activity was low in both cell lines (see Additional file 1) and no production of PIC was detected in culture media of the cell lines (Figure 1d and h). Moreover, a negative value for PIC (N19: untreated: −10.345 nM, IFN-γ- treated: −9.234 nM; N20.1: untreated: −13.345 nM, IFN-γ-treated: −16.356 nM) indicate post-culture concentrations were lower than that in pre-culture conditions. This suggests exogenous PIC, present in serum supplemented media, is taken up and/or catabolized by the oligodendrocytes. Low QUIN concentrations were also observed in the culture media of both cell types (Figure 1d and h). Along with the elevated activity of QPRT seen (see Additional file 1) this suggests an increased uptake and degradation of QUIN by oligodendroglial cells. To address this possibility we next investigated QUIN uptake by N19 and N20.1 cells.
Evidence of exogenous QUIN uptake in oligodendrocytes
Using immunocytochemistry, we have demonstrated the capability of both oligodendrocyte cell lines to take up exogenous QUIN (Figure 2). This indicates QUIN is catabolized intracellularly in a time-dependent manner as fluorescence intensity was directly proportional to the uptake of QUIN.
Quinolinic acid (QUIN) time-course study. N19 (top row) and N20.1 cells (bottom row) treated with specified concentration of QUIN for 0, 30, 60 and 90 minutes. Treated cells were analyzed by immunohistochemistry for QUIN uptake.
We then further assessed the cytotoxic threshold of such QUIN uptake on oligodendroglial cells. A standard curve, using commercial exogenous QUIN (Sigma-Aldrich, St Louis, MO, USA), was produced to assess the level of gliotoxicity by QUIN on N19 and N20.1 cells. This demonstrated a relatively similar level of tolerance in the two cell lines to QUIN (Figure 3a and b). There was a notable trend indicating dose-dependent QUIN toxicity on oligodendrocytes where the LD50 for N19 is 0.5 μM and for N20.1 is 1 μM for 72 hours. These LD50 values were chosen as the reference criteria for subsequent QUIN antagonism studies.
Neutralization of exogenous quinolinic acid (QUIN) with an anti-QUIN monoclonal antibody (mAb). The QUIN toxicity assay was investigated in (a) N19 and (b) N20.1 cells treated with various concentration of QUIN (0, 0.05, 0.1, 0.2, 0.5, 1, 2 and 4 μM) for 24 hours, 48 hours and 72 hours. Lactate dehydrogenase (LDH) activity in culture supernatant was determined. The results were analyzed using linear regression analysis. (c) The schematic diagram showed the treatment conditions of QUIN and QUIN mAb on oligodendroglial cells. The effect of QUIN mAb on QUIN toxicity was examined in (d) N19 and (e) N20.1 cells. The levels of LDH released by oligodendroglial cell lines after 72 hours of incubation with 0.5 μM (N19) and 1 μM (N20.1) of QUIN and varying concentrations of QUIN mAb (30 minutes). The study was performed in triplicate and the error bars indicate SE. ***P < 0.001.
Antagonism of QUIN toxicity with neutralizing anti-QUIN monoclonal antibody (mAb)
Based on the 3 conditions illustrated in Figure 3C and the presence of LD50 concentrations of QUIN for 72 hours (determined in Figure 3a and b), linear regression analysis showed that QUIN toxicity on N19 and N20.1 cell cultures was neutralized by QUIN mAb treatments (Figure 3d and e). We found that treatment with anti-QUIN mAb diluted at 1/50 (14 ng/μL) led to complete protection against QUIN excitotoxicity. The pre-treatment with anti-QUIN mAb group was the least effective of the assessed treatments. Comparing the QUIN-PRE treatment groups between both cell lines, the effect of QUIN-induced cell death at LD50 in N20.1 was more reversible than N19. A validation study was performed to determine the concentration of anti-QUIN mAb used in the study was not toxic to the cells (data not shown).
In addition, we also demonstrated that this strategy worked on endogenous QUIN present in supernatants of IFN-γ-treated BV2 microglial cells containing 0.99 ± 0.1 μM of QUIN (data not shown). The glioprotection observed with the QUIN mAb treatment was found to be statistically different (P < 0.001) in all the treatment groups when compared to control cells for both the cell types (Figure 4). Among the treatment groups for both cell lines, anti-QUIN mAb at 1/100 (7 ng/μL) or higher concentration showed full protective effect from QUIN-induced cell death.
Neutralization of endogenous quinolinic acid (QUIN) with an anti-QUIN monoclonal antibody (mAb). (a) The schematic diagram represents the effect of QUIN mAb on IFN-γ-treated BV2 supernatant in N19 and N20.1 oligodendroglial cell lines. (b) represents the levels of lactate dehydrogenase (LDH) released by oligodendroglial cell lines N19 and N20.1, respectively after 72 hours incubation with IFN-γ-treated BV2 and varying concentrations of QUIN mAb. The study was performed in triplicate and the error bars indicate SE. ***P < 0.001.
Inhibition of immune-induced QUIN toxicity with IDO inhibitors
We tested 4 IDO-1 inhibitors including the natural compound berberine and 1-methyl-tryptophan (1MT) isoforms D, L and DL for their ability to block IDO-1 and so stop QUIN production by BV2 cells. We found that all four inhibitors were able to decrease QUIN induced toxicity in a dose dependent manner (Figure 5). Prior to study, IDO-1 inhibitors were tested for toxicity concentrations on BV2 cells and were well tolerated above 4 μM (data not shown). At 1 μM, the 4 IDO-1 inhibitors were able to bring the level of cytotoxicity under the LD50. At 4 μM, all the IDO-1 inhibitors were able to completely abolish the QUIN-induced cell death in both cell lines. Comparing the percentage of cell death between the 2 cell lines, again we observed that N20.1 cells were slightly more sensitive to QUIN toxicity. A concentration of 0.5 μM was needed to rescue N19 cells to LD50 whereas twice as much inhibitor was required to observe a similar effect in N20.1 cells (P < 0.001). Berberine was the less effective compound in protecting N19 cells whilst DL-1MT was the most effective in protecting the N20.1 cells during QUIN excitotoxicity.
Effect of IDO inhibition on immune induced QUIN toxicity. The schematic bar diagram represents the effect of berberine, D-1MT, L-1MT and DL-1MT (0 to 4 μM) on IFN-γ-treated BV2 supernatant in N19 (a) and N20.1 (b) oligodendroglial cell lines. The levels of lactate dehydrogenase (LDH) released by oligodendroglial cell lines after 72 hours incubation with IFN-γ-treated BV2 supernatant. The study was performed in triplicate and the error bars indicate SE. ***P < 0.001.
The KP profile has been extensively characterized in various human cell types including neurons, astrocytes, microglia, endothelial cells and several other immune cells [14]-[17]. However, the literature regarding the KP in oligodendrocytes is very limited [5]. More importantly the biological function of the KP in oligodendrocytes remains to be delineated. In this study, we showed that murine oligodendrocytic cell lines express all the KP enzymes tested and have a fully functional KP unlike astrocytes, which lack the mid-stream enzyme KMO that converts kynurenine to 3-hydroxykynurenine [15]. However, these data conflict with our previous findings showing that human primary oligodendrocytes lack expression of both IDO-1 and TDO-2 [5]. These discrepancies are likely explained by the fact that KP expression: 1) varies between different species of the same cell type [29]-[31] and; 2) is significantly different between primary cells and their respective cell lines [5]. It is also possible that the conditional transfection of the oligodendrocytes with simian virus 40 T antigens may alter the KP expression [17]. Furthermore, in this study we showed that KP metabolism is different between two cell lines of the same cell type arising from the same species. N20.1 cells display a more active KP mRNA enzyme expression when compared to N19 (Figure 1). The degree of gene up-regulation by IFN-γ was also different: most of the KP enzymes were induced by IFN-γ in N20.1 cells but only limited to TDO-2, KAT-2 and KYNU in N19 cells. The increase of K/T ratio as a result of TDO-2 up-regulation in these 2 mice oligodendrocytic cell lines was also reported by our group in the human oligodendrocytic cell line, MO3.13 [5]. It was also demonstrated in a recent study that TDO-2 is highly up-regulated in immortalized glial cell lines [32].
The expression of KATs and secretion of KYNA by both cell lines suggests that the oligodendrocytes are likely to have neuroprotective functions, especially during neuroinflammatory events. This is reflected by higher expression of KAT-II in the presence of IFN-γ (see Additional file 1). This concurs with current literature pertaining to the mice model where KAT-2 is demonstrated to be the main enzyme associated with KYNA production in the brain [33]. Expression of QPRT corresponding with catabolic rather than anabolic activity of QUIN further supports the neuroprotective role of oligodendrocytes. In addition, this suggests that oligodendrocytes are capable of taking up exogenous QUIN present in the extracellular matrix. They could, therefore, be involved in the detoxification of pathophysiological concentrations of exogenous QUIN, secreted by activated immune cells. Using immunocytostaining, we further showed a rapid uptake of QUIN by oligodendrocytes occurring within 30 minutes then gradually increasing with time (Figure 2). These results are in agreement with the KP metabolic profile found in human primary oligodendrocytes showing an uptake of exogenous QUIN from media and production of neuroprotective KYNA and PIC [5]. We previously showed that QUIN is cytotoxic to both primary human neurons and astrocytes at concentrations as low as 150 nM and 350 nM respectively [10].
As described above, N20.1 cells display a more active KP compared to N19 cells. This suggests that N20.1 cells are likely to have a higher tolerance to QUIN toxicity, that is a higher capacity to catabolize QUIN. Indeed, this was reflected by the LD50 concentration to QUIN toxicity in N20.1 cells with a difference 1-fold higher compared to N19 (Figure 3A). Surprisingly, this was not reflected in the PRE-QUIN treatment group where cells are pre-exposed to QUIN followed by treatment with anti-QUIN mAb (Figure 3D and E). We expected N20.1 cells to have a higher capacity to catabolize QUIN. However, the cell death assay revealed otherwise. It is important to note that the QUIN concentrations based on LD50 used in both cell lines are different. It may imply that QUIN at approximately 1 μM concentration range may exert a more permanent and lasting excitotoxic effect. This is regardless of the cells capacity to take up and catabolize QUIN. An important aspect to further dissect is the short-term and long-term effects of QUIN toxicity on oligodendrocytes. This may have important implications for neurodegeneration and repair in situations where the KP is activated for sustained and prolonged periods of time, such as progressive MS. We have previously shown that low micro molar concentrations of QUIN could exert excitotoxicity chronically in human neurons over a period of 5 weeks [34]. Here, our data support that from a previous study by Cammer et al. [8] showing acute QUIN toxicity in primary rat oligodendrocytes at micro molar concentrations.
It is interesting to speculate that differences in susceptibility to QUIN-mediated toxicity observed between the two cell lines may also reflect their different maturation states and the associated expression of NMDA receptors. The relatively immature N19 cells, with a lower expression of NMDA receptors, experience reduced excitotoxicity in response to QUIN. Furthermore, it is also possible that there are differences in expression of QUIN transporter molecules and, therefore, varying levels of QUIN uptake possible between cell lines. The characterization of NMDA receptors and the capacity of the QUIN uptake transporter certainly warrant further investigation in oligodendrocyte development.
It is apparent that therapeutic strategies targeting QUIN could prevent excitotoxicity and ultimately significantly attenuate neurodegeneration. In this study, we demonstrated that neutralizing QUIN toxicity using an anti-QUIN mAb could prevent oligodendrocyte cell death in vitro. There is still a limited number of studies targeting QUIN toxicity with a neutralizing antibody, despite extensive evidence showing QUIN as a key excitotoxin involved in several neuropathological diseases. This study is the first to propose the use of a mAb to neutralize in vitro QUIN toxicity. Based on our findings, the application of neutralizing QUIN with antibodies could be further developed into monoclonal antibody therapy for neurodegenerative diseases such as MS or amyotrophic lateral sclerosis [35]. Monoclonal antibody therapy has gained popularity over the years including the recently FDA approved natalizumab used in MS treatment [36],[37].
As described above, chronic inflammatory responses can easily trigger cumulative production of pathophysiological concentrations of QUIN by activated monocytic cells such as infiltrating macrophages and microglia. This likely creates an environment in which neuronal cells are highly susceptible to excitotoxicity. Thus, we investigated further how manipulation of the KP during inflammation could affect the survival of oligodendrocytes towards QUIN-induced toxicity. We attempted to limit the production of QUIN from the BV2 microglial cell line by inhibiting IDO-1, since this enzyme is highly inducible by IFN-γ and will be of pathophysiological relevance during inflammation. Our data show that the use of IDO-1 inhibitors (1-MT and berberine) were able to significantly decrease QUIN production by BV-2 cells and subsequently lead to complete abolishment of oligodendrocyte cell death at a concentration of 4 μM (Figure 5). These results represent another relevant therapeutic strategy for MS using KP inhibitors. An earlier study using Ro 61-8048, a KMO inhibitor, on EAE rats showed a significant alleviation of disease progression [21]. In this same study the authors demonstrated that the advantage of KMO inhibition, in comparison with IDO-1, is that they could obtain both a decrease of QUIN synthesis and an increased production of the neuroprotective KYNA in the brain and spinal cord [21].
The use of IDO-1 inhibitors as a treatment for EAE and MS requires further investigation. The positive outcomes of this study are based on IDO-1 inhibition specifically in activated microglial cells. However, there is no IDO-1 inhibitor able to selectively target cell types. For example, inhibiting IDO-1 in astrocytes, the main producers of KYNA [15] would likely be deleterious. Furthermore, some studies have demonstrated that systemic inhibition of IDO-1 in EAE exacerbates the disease [38],[39]. It is likely that the timing of intervention is important in this context: normal KP activation certainly suppresses aberrant immune responses and inhibiting this effect is unwanted in autoimmune disease. However, excessive KP activation produces neurotoxic metabolites, so preventing this dysregulated activation of the KP is clearly neuroprotective. Some KP metabolites have critical roles in the regulation of the T-cell activity responsible for autoimmune disorders [39]-[41]. This further supports the notion that the homeostatic activation of IDO-1 in EAE does have beneficial outcomes. However, its continued inhibition could also have significant negative effects in progressive disease.
It is technically challenging to deliver IDO-1 inhibitors specifically to activated monocytic cells. However, cell-specific targeting might be possible using nanoparticles [42]. Another possibility will be to deliver multiple targets of KP modulators that can limit QUIN production whilst increasing KYNA and PIC. Platten et al. [43] demonstrated that tranilast (Rizaben), a synthetic analog of the KP intermediate 3-HAA, was able to fully abolish EAE by skewing the immunological profile from pro-inflammatory to immunosuppressive [43]. Combination therapy targeting KP enzymes and/or metabolites appears to be a promising therapeutic for EAE and MS but requires further investigation.
In conclusion, our data show that the oligodendrocytes express the major components of KP and, importantly, are capable of catabolizing exogenous QUIN. Therefore, they are likely to play a role in the detoxification of excessive QUIN associated with a chronic immune-activated KP in monocytic cells. More importantly, QUIN is toxic to oligodendrocytes at pathophysiological concentrations. Herein, we demonstrated two potential therapeutic strategies to overcome the toxicity of QUIN by directly neutralizing with anti-QUIN mAb and indirectly with KP inhibitors to limit QUIN production. This work presents novel therapeutic insights into excitotoxicity and neuroprotection for oligodendrocytes and other brain cell types.
Main points
First characterization of the kynurenine pathway (KP) in oligodendrocytes
Oligodendrocytes take up QUIN
QUIN is toxic for oligodendrocytes
Exogenous QUIN toxicity can be neutralized by using a monoclonal antibody against QUIN and specific KP inhibitors leading to increased oligodendrocyte survival
GS performed the experiments and acquired the data. GS, SA and CL analyzed the data. GS, CL and GG designed the study and interpreted the data. GS wrote the manuscript. BB, SJ, CL and GG critically revised the manuscript for important intellectual content. All authors read and approved the final manuscript.
CK and GG are sharing senior authorship.
KP:
kynurenine pathway
KYN:
kynurenine
TRP:
KYNA:
kynurenic acid
PIC:
picolinic acid
QUIN:
mAb:
IDO-1:
TDO-2:
1MT:
1-methyl-tryptophan
This work had been funded by the Australian Research Council, the National Health and Medical Research Council, St Vincent's Clinic Foundation and Peter Duncan Foundation. This study had also partly benefited from the Multiple Sclerosis Research Australia and Trish Foundation. The authors wish to acknowledge the Bioanalytical Mass Spectrometry Facility (BMSF) at UNSW for their kind support, especially Ms Sonia Bustamante for her technical expertise and kind advice with the GC/MS. The authors will also like to thanks Dr. Kuffner, Prof Nicholas Hunt and Dr. Campagnoni for their generous provision of cells.
12974_2014_204_MOESM1_ESM.docx Additional file 1: Expression of kynurenine pathway genes in oligodendroglial cells. Levels of transcripts in (a) N19 and (b) N20.1 oligodendroglial cell line in differentiated state, with or without IFN-γ treatment. Dots represent mean value from four different experiments. The levels of transcripts from the genes encoding are IDO1 (indoleamine 2,3-dioxygenase), TDO2 (tryptophan 2,3-dioxygenase), KAT1 (kynurenine aminotransferase1), KAT2 (kynurenine aminotransferase2), KYNU (kynureninase), ACMSD (aminocarboxymuconate semialdehyde decarboxylase) and QPRT (quinolinate phosphoribosyl transferase). Horizontal line indicates median values. ***P < 0.001, **P < 0.01 and *P < 0.05. (DOCX 155 KB)
12974_2014_204_MOESM2_ESM.docx Additional file 2: Expression of kynurenine pathway protein in oligodendroglial cells. Characterization of the kynurenine pathway in (a) N19 and (b) N20.1 cells by immunohistochemistry. Cells showed positive cytoplasmic staining of TDO2 (N19 and N20.1) and KMO (N20.1 only) after 24 hours IFN-γ treatment. IDO1, QUIN and PIC were absent in this cells. Nuclei stained with DAPI marker. Scale bar 20 μm. RAW 264.7 cells were used as control for positive-staining of IDO1, TDO2, KMO, QUIN and PIC. (DOCX 4 MB)
12974_2014_204_MOESM8_ESM.docx Authors' original file for figure 6
12974_2014_204_MOESM10_ESM.docx Authors' original file for figure 8
Applied Neurosciences Program, Peter Duncan Neurosciences Research Unit, St Vincent's Centre for Applied Medical Research, Sydney, Australia
School of Medical Sciences, Faculty of Medicine, University of New South Wales, Sydney, Australia
Department of Neurology, St Vincent's Hospital, Sydney, Australia
Neurodegenerative diseases Research Group, Australian School of Advanced Medicine, Faculty of Human Sciences, Macquarie University, Sydney, 2109, NSW, Australia
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text:
Home Open Access Tracking Project (OATP) abernard102@gmail.com NLM and Publishers Launch Emergency Access Initiative, Granting Free Access to Books and Journals for Healthcare Professionals Responding to Earthquake in Nepal
NLM and Publishers Launch Emergency Access Initiative, Granting Free Access to Books and Journals for Healthcare Professionals Responding to Earthquake in Nepal
abernard102@gmail.com 2015-04-29
"The National Library of Medicine (NLM) and publisher Emergency Access Initiative (EAI) has been activated to support healthcare professionals working on the response to the earthquake in Nepal. The EAI is a collaborative partnership between NLM and participating publishers to provide free access to full-text from more than 650 biomedical journals and more than 4,000 reference books and online databases to healthcare professionals and libraries affected by disasters. It serves as a temporary collection replacement and/or supplement for libraries affected by disasters that need to continue to serve medical staff and affiliated users. It is also intended for medical personnel responding to the specified disaster. EAI is not an open access collection. It is only intended for those affected by the disaster or assisting the affected population. If you know of a library or organization involved in healthcare efforts in response to the earthquake in Nepal, please let them know of this service. This is the fifth time the EAI has been activated. Previously support was provided following the earthquake and subsequent cholera epidemic in Haiti; flooding in Pakistan; the earthquake and tsunami in Japan; and the Ebola outbreak in West Africa. NLM thanks the numerous participating publishers for their generous support of this initiative: American Academy of Pediatrics, American Association for the Advancement of Science, American Chemical Society, American College of Physicians, American Medical Association, American Society of Health-Systems Pharmacists, American Society for Microbiology Press, B.C. Decker, BMJ, Cambridge University Press, EBSCOHost, Elsevier, FA Davis, Mary Ann Liebert, Massachusetts Medical Society, McGraw-Hill, Merck Publishing, Oxford University Press, People's Medical Publishing House, Springer, University of Chicago Press, Wiley and Wolters Kluwer."
http://www.nlm.nih.gov/news/nepal_earthquake_resources_2015.html
Open Access Tracking Project (OATP) » abernard102@gmail.com
oa.new oa.announcement oa.nlm oa.libraries oa.librarians oa.public_health oa.eai oa.publishers oa.business_models

redpajama_set_name: RedPajamaCommonCrawl


text:
Elected Democratic Officials
CCDP
CCDP Faces — We Are All In This Together
https://youtu.be/9d3uEDRITSYThe CCDP Faces video project was slated to be showcased first in our Cleveland County Fair booth beginning Sept. 30. Then the fair was canceled. But the fair committee elected…
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"Postcards to Swing States" Campaign
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Cleveland County Board of Elections Appointed for 2021—2023 Term
The Cleveland County Board of Elections (BOE) is comprised of five members who are appointed every two years. Four of those members are appointed by the State Board of Elections;…
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"Adlai Stevenson II Was My Distant Cousin" ~Evelyn Morehead Around the time I was 10 years old, my mother began to tell me tidbits about our family tree. The one…
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Biden's First 100 Days When Joe Biden was officially sworn in as President on Jan. 20, 2021, the initial reaction for most Americans was an overwhelming sense of calm. It…
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The North Carolina Democratic Party Welcomes President Biden to Raleigh As Part of "National Month of Action" Today, President Joe Biden is visiting Raleigh in an effort to encourage more…
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North Carolina Has One of the Lowest Unemployment Rates In the Country
May State Jobs Report: North Carolina Has One of the Lowest Unemployment Rates In the Country The U.S. Bureau of Statistics today released the State Employment and Unemployment report for…
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Degory Priest (c. 1579–1621) was a passenger on the voyage of the Mayflower in 1620. He signed the Mayflower Compact.
The name of Digory Priest or Prust is common in Devon, Cornwall and London England.
Priest was a member of the Separatists church. This religion was not allowed in England and many people moved to Leiden Netherlands Priest was one of those families.
Priest was a hat-maker from London. There are many legal documents in Leiden which show that he was born in 1579. In 1611 Priest married a widow named Sarah Vincent. Sarah was a sister of Mayflower passenger Isaac Allerton. Priest's became a citizen of Leiden on November 16, 1615 with Isaac Allerton and Roger Wilson.
On June 28, 1617 Priest asked two men to sign a legal document that said that he had not hit John Cripps but only "touched his Jabot". This was the frill on the front of his shirt.
Leiden records show that Degory Priest was a witness to a statement signed on January 18, 1618. The statement was signed by Tailor Isaac Allerton. It was about the cost of a crimson coat.
On April 9, 1619, Degory Priest and Samuel Lee signed a document for Nicholas Claverly, a Tobacco pipe maker. This document was a promise for Claverly's good behavior. Claverly had arrived in Leiden about 1615 and lived in a house owned by Degory Priest.
Records show that on May 3, 1619 Degory Priest was a witness to a document signed by Richard Tyrill. Priest said that Nicholas Claverly was not connected with the murder of Tyrill's brother John Tyrill.
Priest came alone of the Mayflower. His family came over later in another ship.
Priest left Plymouth, England on 16 September 1620. There were 102 passengers and 30–40 crew. On 19 November 1620, the Mayflower reached land at Cape Cod hook. They landed on November 21. They wrote the Mayflower Compact, which made rules on how they would live and treat each other. Degory was one of the signers. The Mayflower was supposed to land in the Colony of Virginia, but the ship was too damaged and they were forced to land at Cape Cod now called Provincetown Harbor.
William Bradford wrote that seven men died soon after the ship landed. "Digerie Preist" was among them. Bradford said that all these died soon after their arrival in the general sickness. He added that Priest's had his wife and children were sent for because she was Mr. Allerton's sister.
Degory Priest married Sarah Vincent on November 4, 1611. She was a widow. They had two daughters. Their names were Marah and Sarah. Sarah Priest married a second time. His was also a hat-maker from Leiden. Their burial places are unknown.
Priest died early in the first winter, on January 1, 1621 of the "general sickness". He was 42 years of age. He was buried in an unmarked grave. This was the custom that first winter of 1620–1621. His name appears on the Pilgrim Memorial Tomb, located on Cole's Hill in Plymouth.
This page was last changed on 25 September 2014, at 12:43.


FRENCHNANNY – Recruitment agency in Russia - Гувернантки из Англии и Франции. Fille au pair en Russie. English-speaking nannies wishing to work in Russia. Сервис во Франции по-русски. Переводчик на Лазурном берегу.
FRENCHNNANY offers recruitment services for English-speaking nannies wishing to work in Russia. Our agency has now been existing for several years.
Since the "au pair" position is not as commonly known in Russia as in Europe, our goal is to promote its existence and its multiple services and advantages.


My passion is taking care of patients and helping them attain the highest level of health possible.
My passion is taking care of patients and helping them attain the highest level of health possible. I have been in practice as a NP in Cary for over 21 years and I cherish developing long-term relationships with my patients. I have a strong focus on preventative healthcare. I believe in patterning with my patients. It is always a happy day when I see a patient meet a health care goal. I have a holistic approach to health care focusing of nutrition, exercise and emotional health.


Duravis R250 tires from Bridgestone are designed to handle the extreme conditions of commercial hauling, delivering and heavy duty service. Dual sidewall casings construction resists damage from punctures, cuts and abrasions while the steel belted construction maintains performance levels and increases tire life overall. Specially engineered tread and groove patterns increase traction while reducing wear over time.


text:
Cameron est une ville américaine située dans le comté de Marshall en Virginie-Occidentale.

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Selon le recensement de 2010, Cameron compte . La municipalité s'étend sur .

La ville est fondée en 1846 par David McConaughey. Il nomme la ville Cameron, en l'honneur de son ami Samuel Cameron, membre de la Baltimore and Ohio Railroad.

Notes et références

City en Virginie-Occidentale
Comté de Marshall (Virginie-Occidentale)

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Image Title: Astounding Free Standing Wood Burning Fireplace Stylish Design With Regard To Freestanding Decor 12. Post Title: Freestanding Wood Burning Fireplace. Filename: astounding-free-standing-wood-burning-fireplace-stylish-design-with-regard-to-freestanding-decor-12.jpg. Image Dimension: 500 x 500 pixels. Images Format: jpg/jpeg. Publisher/Author: Noel Shanahan. Uploaded Date: Thursday - March 01st. 2018 13:03:15 PM. Category: Architecture. Image Source: architecture.com. Burning Wood LAN To Fireplaces Contemporary Free Freestanding Multi Stylish High Fuel Hearth Fireplace On Stovax Regard 25 2. 15. Stoves Astounding 17. Malm Pertaining Quality Design With Regarding Inserts Prepare Decorations Inside 14. 1. For Closed Remodel Woodburning Standing The Ideas Best Products Country Pinterest 5. Modern Vs Decor 12. In 16. Lancer 11. Stove 4.


Addressing women, Imam Khomeini (r.a.) would say, "You accepted that Women's Day should be the birthday anniversary of Fatima Zahra (s.a.) and this creates a responsibility for you." It was decided that Women's Day should fall on Fatima Zahra's (a.s.) birthday anniversary. What was the significance? It was a symbolic act. The meaning of the symbolic act is that women should move ahead on this straight path. And this path brings about greatness and the highest positions for women. It is the path of piety, modesty and knowledge. It is the path of resistance in different arenas. It is the path of raising children, the path of family life. It is the path of all spiritual virtues and values. Women should move ahead in this direction.
Salam, Beautiful article! I will use this to prove that Islam gives women more rights than white Western Feminism does, which isn't really Feminism- it objectifies women. Islam places importance not on a woman's body but on her intellect and wisdom, so it's only logical to say that Islam is greatly more Feminist than Western "Feminism"! God bless Ayatollah Khomeini and the martyrs. Thank you for writing this piece! Mamnoonam!


James D'Amato and Kat Kuhl of Chicago, Illinois are the inventors of Noisy Person Cards, a party game "designed to help people develop new character voices for role-playing games." Similar to Cards Against Humanity and Apples to Apples, The Noisy Person Cards game asks players to give voices to Character Cards (e.g., hectic, adorable, cackling) by reading Phrase Cards (e.g. "I would gladly pay you Tuesday for a hagburger today." The Noisy Person Cards come with 100 Character Cards and 500 Phrase Cards. James and Kat are pitching their game on The Toy Box (May 19).
Prior to The Toy Box, James and Kat successfully raised more than $45,000 (in one month!) to produce The Noisy Person Cards via a Kickstarter campaign. You can download a free print and play edition on their website. The Toy Box airs Fridays at 8pm on ABC.


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Ken Scott Undergraduate Bursary for Overseas Students at University of Dundee in UK, 2017
Ken Scott Undergraduate Bursary for Overseas Students at University of Dundee in UK, 2017 is open for International Students . The scholarship allows Undergraduate level programm(s) in the field of Law taught at University of Dundee . The deadline of the scholarship is .
The University of Dundee is announcing its Ken Scott bursary to undertake to study Diploma in Professional Legal Practice at Dundee Law School in the Academic year 2016/17. Bursaries are open to UK, EU and overseas students for September 2017 starts.
The University of Dundee is a public research university based in the city and Royal burgh of Dundee on the east coast of the central Lowlands of Scotland.
Degree Level: Bursary is available to pursue undergraduate programme.
Available Subject: Bursary is awarded in the field of Law.
Scholarship Benefits: Bursary of £250 value will be awarded.
Number of Scholarships: One bursary is available.
Eligibility: Applications are invited for the above Bursary which is available to one LLB graduate of The University of Dundee who seeks to undertake to study The Diploma in Professional Legal Practice at Dundee Law School in the Academic year 2016/17. Applicants must have already applied for the Diploma in Professional Legal Practice. The Bursary is for the sum of £250 and will be awarded on the basis of financial need, provided that it is evidenced by a third party. Academic merit will not be taken into account.
Nationality: Students from the following countries are eligible to apply to apply.
UK: England, Northern Ireland, Scotland, Wales
EU: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden
Overseas: Afghanistan, Albania, Algeria, Andorra, Angola, Antigua and Barbuda, Argentina, Armenia, Australia, Azerbaijan, Bahrain, Bangladesh, Barbados, Belarus, Belize, Benin, Bhutan, Bolivia, Bosnia and Herzegovina, Botswana, Brazil, Brunei, Burkina Faso, Burma, Burundi, Cambodia, Cameroon, Canada, Cape Verde, Central African Republic, Chad, Chile, China, Colombia, Comoros, Congo, Congo (Democratic Republic), Costa Rica, Cuba, Djibouti, Dominica, Dominican Republic, East Timor, Ecuador, Egypt, El Salvador, Equatorial Guinea, Eritrea, Ethiopia, Fiji, Gabon, Georgia, Ghana, Grenada, Guatemala, Guinea, Guinea-Bissau, Guyana, Haiti, Honduras, Hong Kong, Iceland, India, Indonesia, Iran, Iraq, Israel, Ivory Coast, Jamaica, Japan, Jordan, Kazakhstan, Kenya, Kiribati, Kosovo, Kuwait, Kyrgyzstan, Laos, Lebanon, Lesotho, Liberia, Libya, Liechtenstein, Macedonia, Madagascar, Malawi, Malaysia, Maldives, Mali, Marshall Islands, Mauritania, Mauritius, Mexico, Micronesia, Moldova, Monaco, Mongolia, Montenegro, Morocco, Mozambique, Namibia, Nauru, Nepal, New Zealand, Nicaragua, Niger, Nigeria, North Korea, Norway, Oman, Pakistan, Palau, Panama, Papua New Guinea, Paraguay, Peru, Philippines, Qatar, Russia, Rwanda, Samoa, San Marino, Sao Tome and Principe, Saudi Arabia, Senegal, Serbia, Seychelles, Sierra Leone, Singapore, Solomon Islands, Somalia, South Africa, South Korea, South Sudan, Sri Lanka, St Kitts and Nevis, St Lucia, St Vincent, Sudan, Suriname, Swaziland, Switzerland, Syria, Tajikistan, Tanzania, Thailand, The Bahamas, The Gambia, Togo, Tonga, Trinidad and Tobago, Tunisia, Turkey, Turkmenistan, Tuvalu, Uganda, Ukraine, United Arab Emirates, United States, Uruguay, Uzbekistan, Vanuatu, Vatican City, Venezuela, Vietnam, Yemen, Zambia, Zimbabwe.
Entrance Requirement: Applicants must have their previous degree.
Test Requirement: No
English Language Requirements: Applicants from outside the home country will often need to meet specific English language/other language requirements in order to be able to study there.
How to Apply: The Bursary is to be administered by Blackadders and the successful applicant will be required to attend a cheque presentation in September 2016. Applications should be made by letter sent by post. Please provide the name of someone from the University (lecturer/advisor) who is familiar with the background to your application and with your financial circumstances.
Deadline: Scholarship application deadline is June 24, 2017.
Ken Scott Undergraduate Bursary for Overseas Students at University of Dundee in UK, 2017 is available to undertake Undergraduate level programs at University of Dundee.
20 Fellowships for Developing Countries Students, 2017
University of Dundee Scholarships
Dundee Scholarships


text:
Jerome Corsi said he's raising money for his legal defense. | AP Photo
Roger Stone ally expecting Mueller indictment
Jerome Corsi, a longtime Stone associate and conspiracy theorist, said Mueller snared him in a 'perjury trap.'
By KYLE CHENEY and DARREN SAMUELSOHN
Updated 11/12/2018 06:32 PM EST
2018-11-12T06:32-0500
Jerome Corsi, an ally of longtime Donald Trump associate Roger Stone, said Monday that he expected to be indicted by special counsel Robert Mueller for perjury.
Corsi, a conspiracy theorist who has been meeting with Mueller's team for two months, said that he spoke to prosecutors for around 40 hours but that talks had "all blown up."
"I'm going to be indicted," he said in a livestreamed video. "I anticipate being indicted."
Reports have suggested that Mueller's interest in Corsi stems from a review of communications between him and Stone in 2016 about an impending release of Clinton campaign emails by WikiLeaks, the transparency activist organization. Mueller is investigating whether any Americans had advance knowledge of the email dump, which dominated the final weeks of the campaign.
Corsi called the impending indictment the result of a "perjury trap," though he emphasized that he wasn't totally sure what the indictment might be for. Corsi's attorney declined to comment.
Corsi also repeatedly emphasized his continued support for Trump's presidency and urged backers to send him money for his legal defense.
"I think my crime, really, is supporting Donald Trump," he said, in between long rants about voter fraud and suggesting that Democrats intend to overthrow Trump.
Democrats vow to tighten the screws on Whitaker
By BRENT D. GRIFFITHS
Corsi said prosecutors showed up at his house on Aug. 28, just before his 72nd birthday, and presented a subpoena. He said he didn't talk to them at the time. But later, with his lawyer present, Corsi said, he met multiple times with Mueller's team in Washington. He recalled prosecutors Jeannie Rhee, Aaron Zelinsky and Andrew Goldstein being present.
Corsi also said he was raising money for his legal defense.
His hourlong remarks meandered among various conspiracy theories, suggesting "criminals" were running the Justice Department and that Democrats were seeking to "overthrow" Trump.
"In prison, if I'm allowed to write, maybe I'll do some fiction about this," he said.
Corsi gained notoriety as the author of a book questioning John Kerry's military service during the 2004 presidential campaign. He also fomented birtherism, a conspiracy theory that Trump ultimately championed as well. Trump once retweeted a Corsi article claiming baselessly that Obama's administration fabricated jobs numbers.
According to an ABC report last month, Corsi was scheduled to appear before the Mueller grand jury on Nov. 2. But reporters at the Washington courthouse that day did not see him enter or leave the building.
During a webcast last Monday, Corsi said he'd been involved "in a really constant basis" with the Mueller investigation for about two months.
"It'd been one of the biggest pushes of my life," he said on the show. Beyond that, Corsi said, he couldn't talk about the special counsel inquiry.
"There will come a time when I can and will talk about it," he said, "but right now I'm going to continue to follow my lawyer's advice and be in a 'no comment' mode."
Trump Russia scandal
Mueller Investigation


text:
4-H Insect Identification Contest Study Guide for Senior 4-H'ers
Youth Projects, Insects, Insect Identification
Insects are our most abundant form of wildlife. There are more species of insects than there are species of all other animals and plants combined! Your backyard is inhabited by hundreds of species of insects, each with its own unique and interesting biology and habit. Learning about wildlife is rewarding and fun, and the first step in learning about any type of wildlife is identifying what you are looking at.
Learning to identify insects by participating in the 4-H insect identification contest can provide lifelong benefits. Not only will you be able to identify pest insects and understand how to control them, but you will also be better prepared for high school and college biology classes. You will be a better naturalist with a greater understanding and appreciation of the complexity of life and the world in which you live.
The objective of the 4-H insect identification contest is for contestants to learn basic biology of insects and related organisms, to be able to identify insects to the order level, and to be able to identify 100 of the more common insects and arachnids by their common name.
Although this study guide is primarily intended to help 4-H'ers prepare for the insect identification contest, it can also be used by high school science teachers to help teach students about insects and insect identification. High school students who enjoy learning about insects and other wildlife might even want to participate in the 4-H insect identification contest.
Reference materials for this contest include the following publications:
This study guide (available online or from your local county Extension office)
Publication 317 4-H Introduction to Entomology (available online or from your local county Extension office)
Publication 2297 The 4-H Entomology Manual (available online or from your local county Extension office)
National Wildlife Federation Field Guide to Insects and Spiders and Related Species of North America by Arthur V. Evans (available through libraries and booksellers). This guide includes general information on the biology of insects, spiders, and related arthropods, along with color photographs and basic information for all the insects listed in this study guide. Other field guides and older editions can also be useful, but if you use such guides, be aware that they may use outdated or alternative order classifications.
The state contest is divided into two parts: specimen identification and a written quiz. Specimens may be pinned, preserved in alcohol, shown in photographs, or shown in visual projections. You will be asked to provide information about the specimen. For example, you may be asked to give either the order or common name of the specimen or to indicate the type of life cycle or type of mouthparts. For the written quiz, you will be asked to provide answers to questions about insect biology and habits. Contestants who know the orders of insects, can identify the insects on the study list by their common name, and are familiar with the information in the 4-H Entomology Manual will do well in the contest.
Realize They Are Not All Insects
The Orders of Insects and Arachnids list and the Insect Identification Study List for Seniors both list a few groups or species of land-dwelling creatures that are not insects but do belong to a closely related group of arthropods. These are included because these other land-dwelling arthropods are usually studied by entomologists and because it is important to be able to recognize these creatures and to know they are not insects.
Pay Attention to Size
When taking close-up photos of insects, the goal is usually to fill the frame with the specimen in order to show as much detail as possible. This means that, in most field guides, a flea and a bumble bee will appear about the same size. However, any good field guide will also give information about the approximate length of mature specimens, and it is important to make note of this. When reading about an insect you have never seen before, it can be helpful to look at a ruler and visualize just how long the insect is. Of course, seeing or collecting actual specimens is the best way to learn the size of a particular insect species!
Also keep in mind that there can be considerable size variation among mature specimens of many species of insects. For example, some bumble bees can be more than three times larger than other specimens of the same species. Insects do not grow once they reach the adult stage, but the nutrition they receive in the immature stage has a great influence on adult size.
Know What You Don't Know
Contestants should recognize that the 100 insects on the study list represent only a small fraction of the insect species in the state. In the senior division, contestants may be presented with a few specimens of insects that are not on the study list. If this is done, you will be told at the beginning of the contest, and you will be told how many "off list" specimens there are. In this case, you need not correctly identify the specimen. An answer of "not on list" or "I do not know" will be counted as correct. If you happen to know the insect and correctly identify it, your answer will also be counted correct, but you will not get extra credit. For example, if a squash bee or sweat bee is presented and you are asked to supply the common name, you should identify it as "not listed" or "do not know" rather than calling it a honey bee. However, if asked to give the order, type of mouthparts, or type of life cycle for an "off list" specimen, you should be able to do so. The reason for including a few "off list" specimens in the identification contest is that, when identifying insects, it is as important to know what you don't know as it is to know what you know.
Pay Attention to Spelling
Contestants should strive to spell common names and order names correctly. Answers that are badly misspelled will be counted wrong, and spelling may be used to break ties.
It is also helpful to understand why the common names of some insects are written as one word, as in "dragonfly," while those of other insects are written as two words, as in "house fly." In general, when an organism really is a member of the group being named, the name is written as two words. If the organism does not really belong to that group, then the name is written as one word. For example, dragonflies, butterflies, and fireflies are not really flies, but house flies and horse flies are. Likewise, hornworms are not really worms, but honey bees really are bees. Also, entomologists use the word "bug" only to refer to a certain subgroup of the order hemiptera: stink bugs, squash bugs, ambush bugs, etc. This is why when lady beetles are referred to as "ladybugs," it is written as one word. To an entomologist, ladybugs are not true bugs, just as antlions are not really lions, and silverfish are not really fish.
The Study List
The study list contains the common names of 100 insects and related arthropods. While some of these names refer to a specific insect, many are names for groups of insects, and there may be dozens, even hundreds, of different species within the group. For example, head louse, monarch butterfly, Colorado potato beetle, and cicada killer are common names for specific insects, but termite, lady beetle, mosquito, and longhorn beetle are names for groups of insects. There are several different species of termites, dozens of different species of lady beetles and mosquitoes, and hundreds of different species of longhorn beetles.
In cases where the listed name represents a group of insects, you are not expected to be able to identify individual species within the groups. However, you should be familiar enough with the characteristics of the listed group to be able to identify a member of the group regardless of which particular species is presented. For example, whether presented with a specimen of a cottonwood borer, a locust borer, or an ivory-marked beetle, the contestant should be able to identify the specimen as a longhorn beetle. This is similar to being able to tell whether a particular desert is a cookie, a pie, or a cake even though you may not know exactly what kind of cookie, pie, or cake it is.
You should be able to identify any specimen to the order level and be able to tell what type of life cycle or mouthparts a specimen has even if it is not on the common names list. For example, if presented with a specimen of an eastern Hercules beetle or any other species of beetle, you should be able to identify it as a member of the order Coleoptera and know that it has chewing mouthparts and a complete life cycle. Likewise, if presented with a caterpillar or any adult moth or butterfly, you should be able to recognize it as a member of the order Lepidoptera.
Refer to National Wildlife Federation Field Guide to Insects and Spiders and Related Species of North America by Arthur V. Evans to become familiar with the listed insects. Most specimens presented in the contest will be species illustrated in this field guide. It will also be helpful to view other photos and illustrations of the listed insects using online sources or other field guides. Collecting or observing actual specimens is an even better way to study!
Learning to identify the different orders of insects is the key to learning how to identify insects. Learning the orders is not nearly as difficult as it might first seem. There are only 31 orders, and eight of these (marked with an * in the order table) are so uncommon that you do not need to know them for the contest. See "Orders of Insects and Arachnids" for a list of the orders you need to know.
The names of many orders end in "ptera," which means wing. Learning the wing characteristics for an order can help you quickly identify most adult insects that belong to that order. For example, moths and butterflies, order Lepidoptera (scale wing), have scales on their wings that rub off on your fingers when you touch them. Beetles, order Coleoptera (sheath winged), have hard, sheath-like front wings. Some order names end in "aptera," which means "without wings." Fleas, which have sucking mouthparts and no wings, are in the order Siphonaptera ("Siphon" for tube or pipe and "aptera" for without wings). See the appropriate pages in the 4-H Entomology Manual and in the front of the suggested field guide for more details about insect orders and order names.
Insects within an order have many traits and habits in common, and if you know which order an insect belongs to, you know a lot of other information about that insect. For example, all Diptera have a complete life cycle, and adult flies have sucking or sponging mouthparts. All Hemiptera have a gradual life cycle and have sucking mouthparts as adults and as nymphs. Knowing the orders really is the key to knowing insects! You do not have to memorize what type of mouthparts and life cycle each species of insect has. You just have to know what order it is in and what type of life cycle and mouthparts are characteristic of that order.
This column indicates only a few of the more common hosts or food sources of the listed insect. Contestants should recognize that most of the listed insects have many more hosts that are not listed. For example, horn flies feed primarily on cattle but will also feed on horses, goats, and many other animals. For contest purposes, contestants should list the hosts indicated on the study list. Hosts listed for Lepidoptera are for the larval stage.
Mouthparts
All insects do not feed in the same way. Some feed on plants by eating the leaves or by boring through fruit, stems, or trunks. Others feed on plants by sucking sap, and still others feed on animals by sucking blood. Moths and butterflies use siphoning mouthparts to suck nectar from flowers through a long thin proboscis. Some flies have sponging/sucking type mouthparts, and some adult insects do not have working mouthparts at all because they do not live very long and do not feed as adults. Knowing what type of mouthparts an insect has can help you know what type of damage it will cause; if the insect is a pest, this information can even help determine which kinds of control will work best.
For purposes of this contest, we will refer to the following five types of mouthparts:
Chewing (C)—Distinguished by a pair of strong mandibles and a smaller pair of maxillae. Characteristic of beetles, caterpillars, and several other orders.
Piercing/sucking (P/S)—Distinguished by an elongated, hollow proboscis used to pierce the plant, animal, or insect being fed upon and suck up sap or blood. Characteristic of the Hemiptera and many flies, such as mosquitoes.
Chewing and sucking (C&S)—Distinguished by having chewing mandibles as well as other mouthparts modified for sucking. Characteristic of bees.
Sponging (SP)—Distinguished by a hollow proboscis with an enlarged sponge-like structure at the end. Characteristic of house flies and some other flies.
Siphoning (SIP)—Distinguished by a long, hollow proboscis that is usually rolled up when not in use. Characteristic of adult moths and butterflies.
See the information on "How Insects Feed— Mouthparts" in the 4-H Entomology Manual for more information on insect mouthparts and how insects feed.
Be aware that some insects have one type of mouthparts as immatures and a completely different type of mouthparts as adults. For example, moths and butterflies have chewing mouthparts as immatures and siphoning mouthparts as adults. Also be aware that some insects have unusual mouthparts that do not fit any of the above categories. For example, immature lacewings and some predatory beetles have hollow mandibles through which they suck the blood of their insect prey. Head lice have sucking mouthparts, but they are not like the piercing/sucking mouthparts of a bed bug. The sucking mouthparts of spiders, ticks, and mites do not fit any of the above categories exactly.
Pest Status or Harmful Stage
The information in this column indicates whether a particular insect or group of insects is considered to be a pest, a beneficial, or neutral (not pests). If the insect is a pest, the harmful stage is indicated. Contestants should recognize that this information is somewhat subjective in nature, and there are often exceptions. For example, cicadas are not usually pests, but large numbers of periodic cicadas can damage fruit trees by causing twigs to break where they insert their eggs. Even though adult females can sting if mishandled, cicada killers are also listed as "not pests" because stings are very rare. Similarly, praying mantids are not listed as being beneficial, even though they are predators of other insects, because they usually prey on insects that are not pests.
When participating in the contest, answer questions based on the information presented in the study guide—even if you are aware of exceptions to the general information given in the study guide. Biology is not always an exact science; it is often a science of generalities and exceptions to those generalities.
Orders of Insects and Arachnids
Type Mouthparts
Class Entognatha
Protura*
proturans
springtails
chewing or sucking
Diplura*
diplurans
Class Insecta
Thysanura
silverfish and firebrats
Microcoryphia*
bristletails
grasshoppers and crickets
Phasmida
Grylloblattodea*
rockcrawlers
Mantophasmatodea*
mantophasmids
Isoptera
Plecoptera
Dermaptera
Embioptera*
web-spinners
Psocoptera
barklice and booklice
Phthiraptera
chewing lice and sucking lice
Zoraptera *
zorapterans
Hemiptera1
true bugs, cicadas, leafhoppers, fulgorids, aphids, whiteflies, scales
piercing/sucking
Ephemeroptera
chewing (immature), none (adult)
Thysanoptera
Neuroptera2
lacewings, antlions, dobsonflies, snakeflies
Strepsiptera*
twisted-wing parasites
Mecoptera
scorpionflies
chewing (immature), piercing/sucking (adult)
chewing (immature), sucking or sponging (adult)
Trichoptera
chewing (immature), sucking (adult)
chewing (immature), siphoning (adult)
ants, bees, and wasps
chewing or chewing and sucking
Common Orders of Arachnids, Class Chelicerata
ticks and mites
shewing
Scorpiones
Pseudoscorpiones
*Orders followed by an asterisk are uncommon and are rarely seen by most amateur entomologists. Specimens from these orders will not be included in the contest.
1Earlier books divide the Hemiptera into two orders, Hemiptera (true bugs) and Homoptera (cicadas, leafhoppers, fulgorids, aphids, whiteflies, and scales).
2Some books, including the Field Guide to Insects and Spiders and Related Species of North America, place dobsonflies and snakeflies in separate orders, but for contest purposes, they are considered to be Neuroptera.
Insect Identification Study List for Seniors
Insect*
Mouth Parts
Not pests
Nymphs and adults
antlion (i)
Neuroptera
sucking (i)
baldfaced hornet
Adults (will sting if disturbed)
barklouse
fungi and lichens
black and yellow mud dauber
Adults (nests can damage equipment)
black carpenter ant
insects and honeydew (nests in wood)
black soldier fly
decaying organic matter
C (i)
Larvae and adults
black swallowtail butterfly (i)
parsley, dill, fennel
Larvae (pests in herb gardens); adults welcome in butterfly gardens
black widow spider
Immatures and adults (bite is venomous)
boll weevil
Larvae and adults (eradicated from Mississippi)
pollen and nectar
Beneficial but will sting if disturbed
algae and aquatic plants
carpenter bee
Adults (bore galleries in wood)
cecropia moth (i)
leaves of many trees
chinch bug
corn, St. Augustine grass
roots of trees
cicada killer
Not pests (adults can sting)
potato and tomato
Immatures and adults (webs are unsightly)
corn earworm (i)
corn, cotton, vegetables
crane fly
grass thatch
cucumber beetle, 12-spotted
general garden feeders
dragonfly (i)
predator, eats mosquitoes
Beneficial: niad and adult
predator (some plant-feeding)
Immatures and adults (can damage rose blooms)
eyed click beetle
field cricket
general plant feeders
fiery searcher
Not a pest
Siphonoptera
golden silk orb weaver
granary weevil
stored grains
green bottle fly
feces and carrion
green lacewing
Beneficial: larvae
green stink bug
plant seeds and fruits
harvestman
predator/scavenger
head louse
Beneficial (but will sting if disturbed)
horned passalus
decaying logs
horn fly
hornworm (i)
tomato, pepper
horse fly
garbage, manure (i)
hummingbird moth
honeysuckle (i)
ichneumon wasp
parasites of other insects
imported fire ant
insects and seeds
Adults (sting if disturbed)
Indian meal moth
grains and dried stored foods
jewel beetles
larvae bore in trees
azalea, eggplant, pyracantha
lady beetle (i)
Beneficial: larvae and adults
leafhopper
most plants
longhorn beetles
luna moth (i)
leaves of hardwood trees
May beetle (i)
roots of grasses (i)
aquatic plants and detritus (i)
Adults (nuisance when numerous)
various plants
Mexican bean beetle
beans, cowpeas
roots of grasses
monarch butterfly (i)
milkweed plants (i)
blood of mammals
praying mantid
pseudoscorpion
red-spotted purple butterfly (i)
willow and poplar
robber fly
scorpionfly
insects and decaying vegetation
Thyanura
cotton clothing, paper
Immatures and adults
soldier beetle
soybean looper (i)
soybeans, other plants
spider mite, two-spotted
many plants
springtail
fungi and decaying vegetation
squash bug
curcubits
squash and pumpkins
stable fly
plant material (i)
striped blister beetle
vegetables and crops
tarnished plant bug
wood, paper
plants, leaves, or flowers
mammals and other animals
tiger beetle
tiger swallowtail butterfly
various trees
true katydid
varied carpet beetle
wool and silk fabrics, feathers and fur
velvet ant
parasites of ground-nesting bees and wasps
viceroy butterfly
willow and other trees
vinegar fly
ripe fruit
walkingstick
Not pests (two-striped walkingsticks can release a spray that causes temporary blindness)
white-fringed beetle
yellow garden spider
(i) = immature (meaning that the host or type of mouthparts listed are for the immature stage)
*In most cases, contestants will be expected to identify adult insects, but when the common name is followed by an (i), you should be able to identify either the adult or the immature stage. Contestants may use the following abbreviations (or may use complete spelling):
For mouthparts: C = chewing; P/S = piercing/sucking; C&S = chewing and sucking; SP = sponging; Sip = siphoning; Sucking = sucking (as in spiders, ticks, and spider mites)
By Blake Layton, PhD, Extension Professor, Entomology and Plant Pathology.
Attract pollinators to gardens in 3 easy steps
Seed treatments lose punch against thrips
Bee health complexity requires scientific solutions
Mississippi farmers, beekeepers work together to protect bees
MSU scientists study tie between insecticides and bee health
Mississippi 4-H Junior Master Gardener Program - Information for Prospective Sponsors
Mississippi 4-H Report Form - Ages 8-11
Mississippi 4-H Report Form - Ages 12-18
Design Your Own Clover
Mississippi 4-H Junior Master Gardener Program - Guidelines for Volunteer Leaders
Crape Myrtle Bark Scale Vol. 5, No. 6
Crane Fly Vol. 5, No. 5
Eastern Subterranean Termite Swarmers Vol. 5, No. 3
Cabbage Root Aphid/Poplar Petiolegall Aphid, Vol. 5, No.2
Red Flour Beetle, Vol. 5, No.1
http://extension.msstate.edu/publications/publications/4-h-insect-identification-contest-study-guide-for-senior-4-hers


Mat sits in an ale house, drinking and relieving the last few hours of his life. The Battle in the Stone; Mat's life was saved by a horde of Trollocs, we'll read his flashback to you. We'll spend a little time talking about Manetheran, and how the Pattern itself has picked the perfect people to save the world from the touch of the Dark One. While brooding to himself, Mat overhears a group of men talking about the Two Rivers. They're merchants, and a few are concerned that there will be no fine Two Rivers tabac to sell next year, because there is a large group of Whitecloaks occupying that region. We all know what happens when the Whitecloaks come to town. Mat returns to the Stone to inform his friends of this news, but when he finds Perrin, the Blacksmith is already packing his things. Their paths are laid out for them, Mat's leads to the Waste, intertwined with Rand's. Perrin's though, leads home.
spoilercon.org Plans are already in the works for next year's WotSpoiler's meet-up.


text:
Q: Android data binding not working with View 'android:tag' property Trying to use the new Android Data Binding in my project, but I'm getting an error when trying to set the 'android:tag' property to some custom variable.
My menu_item.xml file:
<?xml version="1.0" encoding="utf-8"?>
    <layout xmlns:tools="http://schemas.android.com/tools"
        xmlns:android="http://schemas.android.com/apk/res/android">

    <data>

        <variable
            name="menuItem"
            type="com.example.MenuItem" />
    </data>

    <LinearLayout
        android:layout_width="match_parent"
        android:layout_height="match_parent"
        android:orientation="horizontal"
        android:tag="@{menuItem}"
        tools:ignore="UseCompoundDrawables">

        <!--suppress AndroidUnknownAttribute -->
        <ImageView
            android:id="@+id/icon"
            android:layout_width="wrap_content"
            android:layout_height="wrap_content"
            android:imageResource="@{menuItem.itemType.drawableId}" />

        <TextView
            android:id="@+id/displayName"
            android:layout_width="wrap_content"
            android:layout_height="wrap_content"
            android:text="@{menuItem.itemType.displayNameId}" />

    </LinearLayout>
</layout>

My MenuItem class:
public class MenuItem {

    public final ItemType itemType;

    public MenuItem(ItemType itemType) {
        this.itemType = itemType;
    }

}

Part of the genetated MenyItemBinding.java:
public MenuItemBinding(View root) {
        super(root, 0);
        final Object[] bindings = mapBindings(root, 3, sIncludes, sViewsWithIds);
        this.displayName = (android.widget.TextView) bindings[2];
        this.displayName.setTag(null);
        this.icon = (android.widget.ImageView) bindings[1];
        this.icon.setTag(null);
        this.mboundView0 = (android.widget.LinearLayout) bindings[0];
        this.mboundView0.setTag(root.getResources().getString(com.myApp.R.string.@{menuItem}));
        setRootTag(root);
        invalidateAll();
    }

And the error is in the generated class, when trying to set the Tag of the bound view.
Any ideas how to get around this? Preferably, not to use a custom LinearLayout to support this.

A: That is a bug. We haven't tried data binding tags, mostly because tags are special.
When targeting devices pre-ICS, Android data binding takes over the tag of the outermost element of the layout. This tag is used for mostly for binding lifecycle and is used by DataBindingUtil.findBinding() and DataBindingUtil.getBinding().
So, since data binding isn't working on tags, the only work-around is to not supply a tag to your LinearLayout or supply a fixed tag or resource string. If you are targeting ICS and above, it is valid to reassign the tag after binding the layout:
MenuItemBinding binding = MenuItemBinding.inflate(layoutInflater);
binding.getRoot().setTag(menuItem);

You can also create a BindingAdapter for a new attribute:
@BindingAdapter("specialTag")
public static void setSpecialTag(View view, Object value) {
    view.setTag(value);
}

and then use it in your layout:
<LinearLayout
    android:layout_width="match_parent"
    android:layout_height="match_parent"
    android:orientation="horizontal"
    app:specialTag="@{menuItem}"
    tools:ignore="UseCompoundDrawables"/>

This will allow you to use findViewByTag() and all of the other things you expect.
However, this will NOT work if you target Honeycomb and earlier devices. There is no getting around that. You may be tempted to do something like this:
@BindingAdapter("specialTag")
public static void setSpecialTag(View view, Object value) {
    view.setTag(R.id.app_tag, value);
}

You won't be able to use findViewByTag with that approach, but it will store whatever value you want when you use your view. But the reason we don't use ID'd tags with Honeycomb and earlier is that there is a memory leak when using ID'd tags, so don't do it.
I hope this helps. I'll file a bug internally to support data bound android:tags.

redpajama_set_name: RedPajamaStackExchange


text:
Q: Can I implement Google's AdWord server-to-server "installs" conversion tracking from an iOS app? Google's AdWords product supports a server-to-server API.
Give the instructions on that page can I make a call to https://www.googleadservices.com/pagead/conversion/MY_CONVERSION_ID/ from within an iOS app or does my App need to call my server enabling my server to make the call?
If I can do from an iOS app are there ObjC or Swift examples?


Every Wednesday at 5:30 with the KTC Socialites.
Distances range from about a 5K to 6 miles.
Bring a friend! Join 'em afterwords for food and libations!
1st Saturday of the month at 8am! meet at Runners Market. Distance ranges from 3.5 – 6 miles or more!
Runners Market is a proud supporter of the Legacy Parks Foundation! Shop at Runners Market on the first Saturday of each month and a portion of the proceeds goes back to their work to expand, protect, and access our parks, trails, and waterways in East Tennessee!
At Runners Market we are Knoxvillians just like you. We live here, work here, and run here! We are dedicated to providing you, our neighbor, with quality advice and friendly service that can't be found at any big box store in town. We will always do our best to make you feel at home, so come on by!
Our friendly staff is committed to properly fitting you for shoes best suited for your feet and level of activity. Click here for more information on the shoe fitting process and how we can help you run and walk longer in comfortable shoes!
Knoxville is an excellent place to enjoy a run or walk year round. As outdoor enthusiasts, our staff can provide recommendations for runs, walks, and hikes in the Knoxville area. Stop by for directions, maps, and tips on where to go.
Struggling to find a shoe that fits and feels great? Our trained and experienced staff can help! We can fit you for shoes that work with the biomechanics of your feet to ensure you and your feet stay happy!
If you enjoy the thrill of competition or just that feeling of camaraderie at community races, here you will find a comprehensive list of area road and trail races to help you stay on track for your next outing.


Software for macromolecular model-building. Contribute to pemsley/coot development by creating an account on GitHub.
23/10/2018 · Add a garlic clove to the water. Tossing in a garlic clove or two while the water comes to a boil is a great way to add earthy flavor to your hot dogs. You don't even have to peel the garlic; just throw in a clove or two unpeeled.
If you're hitting the tracks of Mount Coot-tha, make sure you have ample water, a good map and don't set off too late in the day. Sometimes the inclines take longer than you think. Sometimes the inclines take longer than you think.
[Coot puts the sidechain back to the original position] Click "Real Space Refine Zone" in the Model/Fit/Refine window 10 . In the graphics window, click on an atom of residue 89A.